Antibodies that bind to integrin alpha 11 and uses thereof

Abstract

The present disclosure provides an antibody or antigen-binding fragment thereof that specifically binds to integrin alpha 11 (ITGA11). The present disclosure also provides methods for treating disorders such as fibrosis, inflammatory disorders, or cancer in a subject. The present disclosure includes related pharmaceutical compositions, polynucleotides, vectors, host cells, production methods, therapeutic methods, diagnostic methods, and kits.

Description

Related Applications

[0001] This Patent Cooperation Treaty application claims priority to U.S. Provisional Patent Application No. 63 / 380,668, filed October 24, 2022, the entire contents of which are incorporated herein by reference. [Background technology]

[0002] Interactions between cells and their extracellular microenvironment are primarily mediated by a family of cell surface receptors known as integrins, which are recognized as key molecules involved in myofibroblast differentiation. Integrins are heterodimeric transmembrane receptors composed of α and β subunits that combine to form 24 different integrin heterodimers. Integrins control cytoskeletal dynamics, thereby influencing many important cellular processes, such as cell adhesion, migration, and differentiation. Integrins also play a key role in the activation of growth factors such as transforming growth factor β (TGFβ). The integrin alpha chain α11 (ITGA11) interacts with the integrin beta chain β1 to form the α11β1 heterodimer. α11β1 selectively binds type I collagen and has been shown to be expressed on the cell surface of cultured fetal and cardiac fibroblasts, as well as activated liver, lung, and kidney myofibroblasts. α11β1 has been shown to be upregulated by TGFβ and regulate the differentiation of embryonic mesenchymal cells on collagen matrices. α11β1 is also involved in tumor growth induction and the metastatic potential of small cell lung cancer cells. ITGA11 is overexpressed in fibrotic tissues, including the lungs of patients with IPF and the kidneys of patients with chronic allograft nephropathy. Therefore, ITGA11 is a target for therapeutic intervention, and there is a need for therapeutic modalities that bind to ITGA11. Summary of the Invention

[0003] The present disclosure provides an antibody or antigen-binding fragment thereof that specifically binds to integrin alpha 11 (ITGA11). The present disclosure also provides a method for treating an ITGA11-associated disorder (such as fibrosis, inflammatory disorder, or cancer) in a subject. The present disclosure also includes related pharmaceutical compositions, polynucleotides, vectors, host cells, production methods, therapeutic methods, diagnostic methods, and kits.

[0004] In one aspect, the present disclosure provides an antibody or antigen-binding fragment thereof that specifically binds to integrin alpha 11 (ITGA11), The antibody or antigen-binding fragment thereof includes: a complementarity determining region (CDR) heavy chain 1 (CDR-H1) comprising the amino acid sequence of GFTFSSYA (SEQ ID NO: 1), GFTFSNAW (SEQ ID NO: 9), GFTFSSYS (SEQ ID NO: 14), GYTFTDYY (SEQ ID NO: 28), GFTFSDYW (SEQ ID NO: 36), or GFMFDTHA (SEQ ID NO: 46); a complementarity-determining region (CDR) heavy chain 2 (CDR-H2) comprising the amino acid sequence of ISGSGGST (SEQ ID NO: 2), ISSSSTI (SEQ ID NO: 15), FDPEDGET (SEQ ID NO: 29), or ISGSGGSI (SEQ ID NO: 74); Complementarity determining region (CDR) heavy chain 3 (CDR-H3) comprising the amino acid sequences of AKDLDWSGHDAFDI (SEQ ID NO: 3), ARDRGYSYSETSNDAFDI (SEQ ID NO: 10), ARGPDLSDYFDY (SEQ ID NO: 16), AKDPRGSGRDDAFDI (SEQ ID NO: 20), AKDPTTMTTDAFDI (SEQ ID NO: 25), ATLDYRGVVYFDY (SEQ ID NO: 30), AKDLLWAARDAFDI (SEQ ID NO: 37), AKQTVTSADDYFDY (SEQ ID NO: 43), ARSGETAGTDYFDY (SEQ ID NO: 48); Complementarity-determining region (CDR) light chain 1 (CDR-L1) comprising the amino acid sequences of QSISSY (SEQ ID NO: 4), QTIGSY (SEQ ID NO: 21), SGSIASNY (SEQ ID NO: 31), QGINDF (SEQ ID NO: 40), and QSVSSSY (SEQ ID NO: 49); Complementarity-determining region (CDR) light chain 2 (CDR-L2) comprising the amino acid sequence of AAS (SEQ ID NO: 5), GAS (SEQ ID NO: 22), or EDK (SEQ ID NO: 32); and Complementarity-determining region (CDR) light chain 3 (CDR-L3) comprising the amino acid sequence of QQTYSTPLT (SEQ ID NO: 6), QQSYSTPFT (SEQ ID NO: 11), QQSYSTPLT (SEQ ID NO: 17), QSYDSSNHWV (SEQ ID NO: 33), or QQDYNSPYT (SEQ ID NO: 50).

[0005] In some embodiments, the antibody or antigen-binding fragment comprises a CDR-H1 comprising the amino acid sequence of GFTFSSYA (SEQ ID NO: 1).

[0006] In some embodiments, the antibody or antigen-binding fragment comprises a CDR-H2 comprising the amino acid sequence of ISGSGGST (SEQ ID NO: 2).

[0007] In some embodiments, the antibody or antigen-binding fragment comprises a CDR-L1 comprising the amino acid sequence of QSISSY (SEQ ID NO: 4).

[0008] In some embodiments, the antibody or antigen-binding fragment comprises a CDR-L2 comprising the amino acid sequence of AAS (SEQ ID NO: 5).

[0009] In some embodiments, the antibody or antigen-binding fragment comprises a CDR-L3 comprising the amino acid sequence of QQSYSTPFT (SEQ ID NO: 11).

[0010] In some embodiments, the antibody or antigen-binding fragment thereof comprises: CDR-H1 comprising the amino acid sequence of GFTFSSYA (SEQ ID NO: 1); CDR-H2 comprising the amino acid sequence of ISGSGGST (SEQ ID NO: 2); CDR-H3 comprising the amino acid sequence of AKDLDWSGHDAFDI (SEQ ID NO: 3); CDR-L1 comprising the amino acid sequence of QSISSY (SEQ ID NO: 4); CDR-L2 comprising the amino acid sequence of AAS (SEQ ID NO: 5); and A CDR-L3 comprising the amino acid sequence of QQTYSTPLT (SEQ ID NO: 6). In some embodiments, the antibody or antigen-binding fragment thereof comprises: a heavy chain variable domain comprising an amino acid sequence having at least 90% (e.g., at least 95%, 96%, 97%, 98%, or 99%) sequence identity to the amino acid sequence of SEQ ID NO:7; and A light chain variable domain comprising an amino acid sequence having at least 90% (e.g., at least 95%, 96%, 97%, 98%, or 99%) sequence identity to the amino acid sequence of SEQ ID NO:8. In some embodiments, the antibody or antigen-binding fragment thereof comprises: a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO:7; and A light chain variable domain comprising the amino acid sequence of SEQ ID NO:8.

[0011] In some embodiments, the antibody or antigen-binding fragment thereof comprises: CDR-H1 comprising the amino acid sequence of GFTFSNAW (SEQ ID NO: 9); CDR-H2 comprising the amino acid sequence of ISGSGGST (SEQ ID NO: 2); CDR-H3 comprising the amino acid sequence of ARDRGYSYSETSNDAFDI (SEQ ID NO: 10); CDR-L1 comprising the amino acid sequence of QSISSY (SEQ ID NO: 4); CDR-L2 comprising the amino acid sequence of AAS (SEQ ID NO: 5); and A CDR-L3 comprising the amino acid sequence of QQSYSTPFT (SEQ ID NO: 11). In some embodiments, the antibody or antigen-binding fragment thereof comprises: a heavy chain variable domain comprising an amino acid sequence having at least 90% (e.g., at least 95%, 96%, 97%, 98%, or 99%) sequence identity to the amino acid sequence of SEQ ID NO: 12; and A light chain variable domain comprising an amino acid sequence having at least 90% (e.g., at least 95%, 96%, 97%, 98%, or 99%) sequence identity to the amino acid sequence of SEQ ID NO:13. In some embodiments, the antibody or antigen-binding fragment thereof comprises: a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 12; and A light chain variable domain comprising the amino acid sequence of SEQ ID NO: 13.

[0012] In some embodiments, the antibody or antigen-binding fragment thereof comprises: CDR-H1 comprising the amino acid sequence of GFTFSSYS (SEQ ID NO: 14); CDR-H2 comprising the amino acid sequence of ISSSSSTI (SEQ ID NO: 15); CDR-H3 comprising the amino acid sequence of ARGPDLSDYFDY (SEQ ID NO: 16); CDR-L1 comprising the amino acid sequence of QSISSY (SEQ ID NO: 4); CDR-L2 comprising the amino acid sequence of AAS (SEQ ID NO: 5); and CDR-L3 comprising the amino acid sequence of QQSYSTPLT (SEQ ID NO: 17). In some embodiments, the antibody or antigen-binding fragment thereof comprises: a heavy chain variable domain comprising an amino acid sequence having at least 90% (e.g., at least 95%, 96%, 97%, 98%, or 99%) sequence identity to the amino acid sequence of SEQ ID NO: 12; and A light chain variable domain comprising an amino acid sequence having at least 90% (e.g., at least 95%, 96%, 97%, 98%, or 99%) sequence identity to the amino acid sequence of SEQ ID NO:13. In some embodiments, the antibody or antigen-binding fragment thereof comprises: a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 12; and A light chain variable domain comprising the amino acid sequence of SEQ ID NO: 13.

[0013] In some embodiments, the antibody or antigen-binding fragment thereof comprises: CDR-H1 comprising the amino acid sequence of GFTFSSYA (SEQ ID NO: 1); CDR-H2 comprising the amino acid sequence of ISGSGGST (SEQ ID NO: 2); CDR-H3 comprising the amino acid sequence of AKDPRGSGRDDAFDI (SEQ ID NO: 20); CDR-L1 comprising the amino acid sequence of QTIGSY (SEQ ID NO: 21); CDR-L2 comprising the amino acid sequence of GAS (SEQ ID NO: 22); and A CDR-L3 comprising the amino acid sequence of QQSYSTPFT (SEQ ID NO: 11). In some embodiments, the antibody or antigen-binding fragment thereof comprises: a heavy chain variable domain comprising an amino acid sequence having at least 90% (e.g., at least 95%, 96%, 97%, 98%, or 99%) sequence identity to the amino acid sequence of SEQ ID NO:23; and A light chain variable domain comprising an amino acid sequence having at least 90% (e.g., at least 95%, 96%, 97%, 98%, or 99%) sequence identity to the amino acid sequence of SEQ ID NO:24. In some embodiments, the antibody or antigen-binding fragment thereof comprises: a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 23; and A light chain variable domain comprising the amino acid sequence of SEQ ID NO:24.

[0014] In some embodiments, the antibody or antigen-binding fragment thereof comprises: CDR-H1 comprising the amino acid sequence of GFTFSSYA (SEQ ID NO: 1); CDR-H2 comprising the amino acid sequence of ISGSGGST (SEQ ID NO: 2); CDR-H3 comprising the amino acid sequence of AKDPTTMTTDAFDI (SEQ ID NO: 25); CDR-L1 comprising the amino acid sequence of QSISSY (SEQ ID NO: 4); CDR-L2 comprising the amino acid sequence of AAS (SEQ ID NO: 5); and A CDR-L3 comprising the amino acid sequence of QQSYSTPFT (SEQ ID NO: 11). In some embodiments, the antibody or antigen-binding fragment thereof comprises: a heavy chain variable domain comprising an amino acid sequence having at least 90% (e.g., at least 95%, 96%, 97%, 98%, or 99%) sequence identity to the amino acid sequence of SEQ ID NO:26; and A light chain variable domain comprising an amino acid sequence having at least 90% (e.g., at least 95%, 96%, 97%, 98%, or 99%) sequence identity to the amino acid sequence of SEQ ID NO:27. In some embodiments, the antibody or antigen-binding fragment thereof comprises: a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 26; and A light chain variable domain comprising the amino acid sequence of SEQ ID NO:27.

[0015] In some embodiments, the antibody or antigen-binding fragment thereof comprises: CDR-H1 comprising the amino acid sequence of GYTFTDYY (SEQ ID NO: 28); CDR-H2 comprising the amino acid sequence of FDPEDGET (SEQ ID NO: 29); CDR-H3 comprising the amino acid sequence of ATLDYRGVVYFDY (SEQ ID NO: 30); CDR-L1 comprising the amino acid sequence of SGSIASNY (SEQ ID NO: 31); CDR-L2 comprising the amino acid sequence of EDK (SEQ ID NO: 32); and A CDR-L3 comprising the amino acid sequence of QSYDSSNHWV (SEQ ID NO: 33). In some embodiments, the antibody or antigen-binding fragment thereof comprises: a heavy chain variable domain comprising an amino acid sequence having at least 90% (e.g., at least 95%, 96%, 97%, 98%, or 99%) sequence identity to the amino acid sequence of SEQ ID NO: 34; and A light chain variable domain comprising an amino acid sequence having at least 90% (e.g., at least 95%, 96%, 97%, 98%, or 99%) sequence identity to the amino acid sequence of SEQ ID NO:35. In some embodiments, the antibody or antigen-binding fragment thereof comprises: a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 34; and A light chain variable domain comprising the amino acid sequence of SEQ ID NO: 35.

[0016] In some embodiments, the antibody or antigen-binding fragment thereof comprises: CDR-H1 comprising the amino acid sequence of GFTFSDYW (SEQ ID NO: 36); CDR-H2 comprising the amino acid sequence of ISGSGGST (SEQ ID NO: 2); CDR-H3 comprising the amino acid sequence of AKDLLWAARDAFDI (SEQ ID NO: 37); CDR-L1 comprising the amino acid sequence of QSISSY (SEQ ID NO: 4); CDR-L2 comprising the amino acid sequence of AAS (SEQ ID NO: 5); and A CDR-L3 comprising the amino acid sequence of QQSYSTPFT (SEQ ID NO: 11). In some embodiments, the antibody or antigen-binding fragment thereof comprises: a heavy chain variable domain comprising an amino acid sequence having at least 90% (e.g., at least 95%, 96%, 97%, 98%, or 99%) sequence identity to the amino acid sequence of SEQ ID NO: 38; and A light chain variable domain comprising an amino acid sequence having at least 90% (e.g., at least 95%, 96%, 97%, 98%, or 99%) sequence identity to the amino acid sequence of SEQ ID NO:39. In some embodiments, the antibody or antigen-binding fragment thereof comprises: a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 38; and A light chain variable domain comprising the amino acid sequence of SEQ ID NO: 39.

[0017] In some embodiments, the antibody or antigen-binding fragment thereof comprises: CDR-H1 comprising the amino acid sequence of GFTFSSYA (SEQ ID NO: 1); CDR-H2 comprising the amino acid sequence of ISGSGGST (SEQ ID NO: 2); CDR-H3 comprising the amino acid sequence of AKDLLWAARDAFDI (SEQ ID NO: 37); CDR-L1 comprising the amino acid sequence of QGINDF (SEQ ID NO: 40); CDR-L2 comprising the amino acid sequence of AAS (SEQ ID NO: 5); and A CDR-L3 comprising the amino acid sequence of QQSYSTPLT (SEQ ID NO: 17). In some embodiments, the antibody or antigen-binding fragment thereof comprises: a heavy chain variable domain comprising an amino acid sequence having at least 90% (e.g., at least 95%, 96%, 97%, 98%, or 99%) sequence identity to the amino acid sequence of SEQ ID NO:41; and A light chain variable domain comprising an amino acid sequence having at least 90% (e.g., at least 95%, 96%, 97%, 98%, or 99%) sequence identity to the amino acid sequence of SEQ ID NO:42. In some embodiments, the antibody or antigen-binding fragment thereof comprises: a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 41; and A light chain variable domain comprising the amino acid sequence of SEQ ID NO:42.

[0018] In some embodiments, the antibody or antigen-binding fragment thereof comprises: CDR-H1 comprising the amino acid sequence of GFTFSNAW (SEQ ID NO: 9); CDR-H2 comprising the amino acid sequence of ISGSGGST (SEQ ID NO: 2); CDR-H3 comprising the amino acid sequence AKQTVTSADDYFDY (SEQ ID NO: 43); CDR-L1 comprising the amino acid sequence of QSISSY (SEQ ID NO: 4); CDR-L2 comprising the amino acid sequence of AAS (SEQ ID NO: 5); and A CDR-L3 comprising the amino acid sequence of QQSYSTPFT (SEQ ID NO: 11). In some embodiments, the antibody or antigen-binding fragment thereof comprises: a heavy chain variable domain comprising an amino acid sequence having at least 90% (e.g., at least 95%, 96%, 97%, 98%, or 99%) sequence identity to the amino acid sequence of SEQ ID NO:44; and A light chain variable domain comprising an amino acid sequence having at least 90% (e.g., at least 95%, 96%, 97%, 98%, or 99%) sequence identity to the amino acid sequence of SEQ ID NO:45. In some embodiments, the antibody or antigen-binding fragment thereof comprises: a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO:44; and A light chain variable domain comprising the amino acid sequence of SEQ ID NO:45.

[0019] In some embodiments, the antibody or antigen-binding fragment thereof comprises: CDR-H1 comprising the amino acid sequence of GFMFDTHA (SEQ ID NO: 46); CDR-H2 comprising the amino acid sequence of ISGSGGSI (SEQ ID NO: 47); CDR-H3 comprising the amino acid sequence of ARSGETAGTDYFDY (SEQ ID NO: 48); CDR-L1 comprising the amino acid sequence of QSVSSSY (SEQ ID NO: 49); CDR-L2 comprising the amino acid sequence of GAS (SEQ ID NO: 22); and A CDR-L3 comprising the amino acid sequence of QQDYNSPYT (SEQ ID NO: 50). In some embodiments, the antibody or antigen-binding fragment thereof comprises: a heavy chain variable domain comprising an amino acid sequence having at least 90% (e.g., at least 95%, 96%, 97%, 98%, or 99%) sequence identity to the amino acid sequence of SEQ ID NO:51; and A light chain variable domain comprising an amino acid sequence having at least 90% (e.g., at least 95%, 96%, 97%, 98%, or 99%) sequence identity to the amino acid sequence of SEQ ID NO:52. In some embodiments, the antibody or antigen-binding fragment thereof comprises: a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 51; and A light chain variable domain comprising the amino acid sequence of SEQ ID NO: 52.

[0020] In some embodiments, the antibody or antigen-binding fragment thereof comprises a heavy chain variable domain comprising an amino acid sequence having at least 90% (e.g., at least 95%, 96%, 97%, 98%, 99%, or 100%) sequence identity to the amino acid sequence of any one of SEQ ID NOs: 7, 12, 18, 23, 26, 34, 38, 41, 44, or 51.

[0021] In some embodiments, the antibody or antigen-binding fragment thereof comprises a light chain variable domain comprising an amino acid sequence having at least 90% (e.g., at least 95%, 96%, 97%, 98%, 99%, or 100%) sequence identity to the amino acid sequence of any one of SEQ ID NOs: 8, 13, 19, 24, 35, 39, 42, 45, or 52.

[0022] In some embodiments, the antibody or antigen-binding fragment thereof is selected from the group consisting of a monoclonal antibody or antigen-binding fragment thereof, a polyclonal antibody or antigen-binding fragment thereof, a human antibody or antigen-binding fragment thereof, a humanized antibody or antigen-binding fragment thereof, a primatized antibody or antigen-binding fragment thereof, a bispecific antibody or antigen-binding fragment thereof, a multispecific antibody or antigen-binding fragment thereof, a dual variable immunoglobulin domain, a monovalent antibody or antigen-binding fragment thereof, a chimeric antibody or antigen-binding fragment thereof, a single-chain Fv molecule (scFv), a diabody, a triabody, a nanobody, an antibody-like protein scaffold, a domain antibody, an Fv fragment, a Fab fragment, a F(ab')2 molecule, and a tandem scFv (taFv).

[0023] In some embodiments, the antibody or antigen-binding fragment thereof is a human antibody, a humanized antibody, or a chimeric antibody, or an antigen-binding fragment thereof.

[0024] In some embodiments, the antibody or antigen-binding fragment thereof specifically binds to the heterodimer of ITGA11 and integrin β1 (ITGA11B1).

[0025] In another aspect, the present disclosure provides a polynucleotide encoding any of the antibodies or antigen-binding fragments thereof described herein.

[0026] In another aspect, the present disclosure provides a vector comprising a polynucleotide described herein (e.g., a polynucleotide encoding any of the antibodies or antigen-binding fragments thereof described herein). In some embodiments, the vector is an expression vector (e.g., a eukaryotic expression vector or a viral vector, such as a viral vector selected from the group consisting of adenovirus (Ad), retrovirus, poxvirus, adeno-associated virus, baculovirus, herpes simplex virus, and vaccinia virus).

[0027] In another aspect, the present disclosure provides a host cell comprising the vector described herein. In some embodiments, the host cell expresses an antibody or antigen-binding fragment thereof described herein.

[0028] In another aspect, the present disclosure provides a pharmaceutical composition comprising an antibody or antigen-binding fragment thereof described herein, a polynucleotide described herein, a vector described herein, or a host cell described herein, and a pharmaceutically acceptable carrier or excipient.

[0029] In another aspect, the present disclosure provides a kit comprising an agent selected from an antibody or antigen-binding fragment thereof described herein, a polynucleotide described herein, a vector described herein, or a host cell described herein, or a pharmaceutical composition described herein.

[0030] In another aspect, the present disclosure provides a method of treating a subject having or at risk of developing a disorder, the method comprising administering to the subject an antibody or antigen-binding fragment thereof described herein, a polynucleotide described herein, a vector described herein, or a host cell described herein, or a pharmaceutical composition described herein.

[0031] In some embodiments, the disorder is fibrosis. In some embodiments, the fibrosis is selected from the group consisting of liver fibrosis (e.g., fibrosis associated with cirrhosis (e.g., alcohol-induced cirrhosis, viral-induced cirrhosis, cirrhosis after hepatitis C, and primary biliary cirrhosis), schistosomiasis, cholangitis (e.g., sclerosing cholangitis), and autoimmune-induced hepatitis); renal fibrosis (e.g., tubulointerstitial fibrosis, scleroderma, diabetic nephritis, and glomerulonephritis); dermal fibrosis (e.g., scleroderma, hypertrophic and keloid scarring, nephrogenic fibrosing dermatosis, and burns); myelofibrosis; neurofibromatosis; fibromas; intestinal fibrosis, and fibrous adhesions due to surgical procedures); cardiac fibrosis (e.g., fibrosis associated with myocardial infarction); vascular fibrosis (e.g., arterial restenosis after angioplasty and atherosclerosis). ocular fibrosis (e.g., fibrosis associated with cataract surgery, proliferative vitreoretinopathy, and retroorbital fibrosis); myelofibrosis (e.g., idiopathic myelofibrosis and drug-induced myelofibrosis); pulmonary fibrosis (e.g., interstitial pulmonary fibrosis); glomerulonephritis; heart failure (ischemic and non-ischemic); scleroderma; excessive scar tissue after surgery or device insertion, trauma or burns, progressive kidney disease, valvular heart disease, hypertensive heart disease, articular and periarticular fibrosis, myelofibrosis, ocular fibrosis / vitreous fibrosis, intestinal fibrosis and stenosis, peritoneal and retroperitoneal fibrosis, pancreatic fibrosis, nephrogenic systemic fibrosis, and primary sclerosing cholangitis. Pathological matrix elaboration also plays a role in fibroproliferative tumor progression and metastasis.

[0032] In some embodiments, the disorder is an inflammatory disorder. In some embodiments, the inflammatory disorder is selected from the group consisting of asthma (e.g., allergic asthma, exercise-induced asthma, aspirin-sensitive / exacerbated asthma, atopic asthma, severe asthma, mild asthma, moderate-to-severe asthma, corticosteroid-naive asthma, chronic asthma, corticosteroid-resistant asthma, corticosteroid-refractory asthma, newly diagnosed and untreated asthma, smoking-induced asthma, corticosteroid-uncontrolled asthma, etc.); airway inflammation, airway hyperreactivity, airway hyperresponsiveness, and the like. hyperresponsiveness), rhinosinusitis, rhinosinusitis with polyps, nasal polyps; arthritis (e.g., osteoarthritis, rheumatoid arthritis, collagen-induced arthritis, arthritis as a result of injury, etc.); eosinophilic inflammation, mast cell-mediated inflammatory diseases, sepsis, septic shock, seronegative enthesitis / arthritis (SEA) syndrome, osteoporosis, eosinophilic esophagitis, scleroderma, dermatitis, atopic dermatitis, allergic rhinitis, bullous pemphigoid, urticaria (e.g., chronic urticaria), inflammation of cartilage, polymyalgia rheumatica, polyarteritis nodosa nodossa, Wegener's granulomatosis, Behçet's disease, myositis, polymyositis, dermatomyolitis, dermatomyositis, vasculitis, arteritis, diabetic nephropathy, interstitial cystitis, graft-versus-host disease (GVHD), inflammatory conditions of the gastrointestinal tract (e.g., inflammatory bowel disease (IBD), ulcerative colitis (UC), Crohn's disease (CD); enteritis (e.g., enteritis due to environmental insults (e.g., due to or related to therapeutic regimens such as chemotherapy, radiation therapy), infectious enteritis, ischemic enteritis, collagenous or lymphocytic colitis, necrotizing enterocolitis; enteritis in conditions such as chronic granulomatous disease or celiac disease; food allergies, gastritis, infectious gastritis or enteritis (e.g., Helicobacter pylori-infected chronic active gastritis), and other forms of gastrointestinal inflammation due to infectious agents);and inflammatory lung pathologies (e.g., chronic obstructive pulmonary disease (COPD), eosinophilic lung inflammation, infection-induced lung pathologies (viral (e.g., influenza, parainfluenza, rotavirus, human metapneumovirus, and respiratory syncytial virus) infections, bacterial infections, fungal infections (e.g., Aspergillus), parasitic infections, or prion infections); allergen-induced lung pathologies, pollutant-induced lung pathologies (e.g., asbestosis, silicosis, or beryllium poisoning), gastric aspiration-induced lung pathologies, immune-modulating selected from nodal abnormalities, genetically predisposed inflammatory conditions (such as cystic fibrosis), trauma-induced pulmonary conditions (e.g., ventilator injury), emphysema, bronchitis, sarcoidosis, histiocytosis, lymphangioleiomyomatosis, acute lung injury, acute respiratory distress syndrome, chronic lung disease, bronchopulmonary dysplasia, pneumonia (e.g., community-acquired pneumonia, hospital-acquired pneumonia, ventilator-associated pneumonia, viral pneumonia, bacterial pneumonia, and severe pneumonia), airway exacerbation, and those associated with acute respiratory distress syndrome (ARDS);

[0033] In some embodiments, the disorder is cancer, hi some embodiments, the cancer is selected from breast cancer, colorectal cancer, liver cancer, kidney cancer, liver cancer, lung cancer, pancreatic cancer, cancer of the gastrointestinal tract, melanoma, ovarian cancer, prostate cancer, cervical cancer, bladder cancer, glioblastoma, head and neck cancer, and bile duct cancer.

[0034] definition To aid in understanding this disclosure, several terms are defined below. Terms defined herein have the meanings commonly understood by one of ordinary skill in the art relevant to this disclosure. Terms such as "a," "an," and "the" are not intended to refer exclusively to a singular item, but include general types of embodiments that may be used for illustration. While the terminology herein is used to describe particular embodiments of the disclosure, their use does not limit the disclosure, except as set forth in the claims.

[0035] As used herein, the term "about" refers to a value that is within 10% of the stated value. For example, the term "about 5 nM" refers to a range of 4.5 nM to 5.5 nM.

[0036] Any value stated herein in a range of values ​​includes both the upper and lower limits, as well as any value included between the upper and lower limits.

[0037] As used herein, the term "antibody" (Ab) refers to an immunoglobulin molecule that specifically binds to or is immunologically reactive with a particular antigen, including polyclonal antibodies, monoclonal antibodies, and genetically engineered or modified antibodies, including, but not limited to, chimeric antibodies, humanized antibodies, primatized antibodies, heteroconjugate antibodies (e.g., bispecific, trispecific, and tetraspecific antibodies, diabodies, triabodies, and tetrabodies), and antigen-binding fragments of antibodies (e.g., Fab', F(ab')2, Fab, Fv, rgG, and scFv fragments). Furthermore, unless otherwise specified, the term "monoclonal antibody" (mAb) is intended to include both intact molecules and antibody fragments (e.g., Fab and F(ab')2 fragments) that are capable of specifically binding to a target protein. Fab and F(ab')2 fragments lack the Fc fragment of intact antibody, clear more rapidly from an animal's circulation, and may have less non-specific tissue binding than intact antibodies (see Wahl et al., J. Nucl. Med. 24:316, 1983; this reference is incorporated herein by reference).

[0038] As used herein, the term "antigen-binding fragment" refers to one or more fragments of an antibody that retain the ability to specifically bind to a target antigen (e.g., as measured by binding affinity). The antigen-binding function of an antibody can be performed by a fragment of a complete antibody. The antibody fragment can be a Fab, F(ab')2, scFv, SMIP, diabody, triabody, affibody, nanobody, aptamer, bispecific, dual-binding bispecific, mAb pair, or domain antibody. Examples of binding fragments encompassed by the term "antigen-binding fragment" of an antibody are: (i) a Fab fragment, which is a monovalent fragment consisting of the VL, VH, CL, and CH1 domains; (ii) F(ab′)2 fragment, a bivalent fragment containing two Fab fragments linked by a disulfide bridge at the hinge region; (iii) an Fd fragment consisting of the VH and CH1 domains; (iv) an Fv fragment consisting of the VL and VH domains of a single arm of an antibody; (v) dAbs containing VH and VL domains; (vi) a dAb fragment consisting of a VH domain (Ward et al., Nature 341:544-546, 1989); (vii) dAbs consisting of VH or VL domains; (viii) an isolated complementarity-determining region (CDR); and (ix) a combination of two or more isolated CDRs, optionally joined by a synthetic linker; Including, but not limited to: Furthermore, although the two domains of an Fv fragment, VL and VH, are encoded by different genes, they can be joined using recombinant techniques by a linker that allows them to be combined into a single protein chain in which the VL and VH regions pair to form a monovalent molecule (known as a single-chain Fv (scFv); see, e.g., Bird et al., Science 242:423-426, 1988; and Huston et al., Proc. Natl. Acad. Sci. USA 85:5879-5883, 1988). These antibody fragments can be obtained using conventional techniques known to those of skill in the art, and the fragments can be screened for utility in the same manner as intact antibodies. Antigen-binding fragments can be produced by recombinant DNA techniques, enzymatic or chemical cleavage of intact immunoglobulins, or, in some embodiments, by chemical peptide synthesis means known in the art.

[0039] As used herein, the term "binding affinity" refers to the strength of all non-covalent interactions between a single binding site of a molecule (e.g., an antibody) and its binding partner (e.g., an antigen or antigenic peptide). Unless otherwise specified, "binding affinity" as used herein refers to intrinsic binding affinity, which reflects the specific interaction between members of a binding pair (e.g., an antibody and an antigen). The affinity of molecule X for its partner Y can generally be expressed by the dissociation constant (Kd). Affinity can be measured by standard methods known in the art, including those described herein. Low affinity complexes generally contain antibodies that tend to dissociate easily from antigens, while high affinity complexes generally contain antibodies that tend to remain bound to antigens for longer periods of time.

[0040] As used herein, the term "chimeric" antibody refers to an antibody having variable domain sequences (e.g., CDR sequences) derived from the immunoglobulin of a source organism (such as a rat or mouse) and constant regions derived from the immunoglobulin of a different organism (e.g., human, other primate, pig, goat, rabbit, hamster, cat, dog, guinea pig, members of the bovine family (such as cow, bison, buffalo, elk, and yak, among others), cow, sheep, horse, or bison, among others). Methods for making chimeric antibodies are known in the art. See, e.g., Morrison, 1985, Science 229(4719):1202-7; Oi et al., 1986, BioTechniques 4:214-221; Gillies et al., 1985, J. Immunol. Methods 125:191-202; U.S. Patent Nos. 5,807,715, 4,816,567, and 4,816,397; these references are incorporated herein by reference.

[0041] As used herein, the term "complementarity-determining region (CDR)" refers to the hypervariable regions found in both the light- and heavy-chain variable domains. The more highly conserved portions of the variable domains are called framework regions (FRs). As understood in the art, the amino acid positions defining the hypervariable regions of an antibody can vary depending on the context and the various definitions known in the art. Some positions within a variable domain may be considered hybrid hypervariable positions; these positions may be considered within a hypervariable region according to one set of criteria, while others are considered outside a hypervariable region according to a different set of criteria. One or more of these positions may also be found in an extended hypervariable region. The antibodies described herein may contain modifications at these hybrid hypervariable positions. Each naturally occurring heavy and light chain variable domain contains four framework regions, primarily in a β-sheet configuration, connected by three CDRs, which form connecting loops and, in some cases, form part of the β-sheet structure. The CDRs of each chain, together with the CDRs of the other antibody chain, are held in close proximity to each other by the FR regions in the order FR1-CDR1-FR2-CDR2-FR3-CDR3-FR4, and contribute to the formation of the antibody target binding site (see Kabat et al., Sequences of Proteins of Immunological Interest (National Institute of Health, Bethesda), Md. 1987; this reference is incorporated herein by reference). In this specification, the numbering of amino acid residues in immunoglobulins is done according to the immunoglobulin amino acid residue numbering system of Kabat et al., unless otherwise specified.

[0042] As used herein, the terms "conservative mutation," "conservative substitution," or "conservative amino acid substitution" refer to the substitution of one or more different amino acids for one or more amino acids that exhibit similar physicochemical properties, such as polarity, electrostatic charge, and steric bulk. These properties are summarized for each of the 20 naturally occurring amino acids in Table 1 below.

[0043] [Table 1]

[0044] From this table it can be seen that conservative amino acid families include, for example, (i) G, A, V, L, I, P, and M; (ii) D and E; (iii) C, S and T; (iv) H, K and R; (v) N and Q; and (vi) F, Y and W. A conservative mutation or substitution is therefore one that replaces one amino acid with a member of the same amino acid family (e.g., substitution of Ser for Thr or Lys for Arg).

[0045] Amino acid substitutions are indicated by the following convention: Amino acid sequences may be represented herein using (AA1)(N)(AA2), where "AA1" represents the amino acid normally present at a particular site within the amino acid sequence, "N" represents the residue number within the amino acid sequence at which the substitution occurs, and "AA2" represents the amino acid present in the amino acid sequence after the substitution is effected. For example, the designation "C232S" in reference to an antibody hinge region, such as an IgG2 antibody hinge region, refers to the substitution of a native cysteine ​​residue with a serine residue at amino acid residue 232 of the hinge amino acid sequence shown. Similarly, the designation "C233S" in reference to an antibody hinge region, such as an IgG2 antibody hinge region, refers to the substitution of a native cysteine ​​residue with a serine residue at amino acid residue 233 of the hinge amino acid sequence shown.

[0046] As used herein, the term "conjugate" refers to a compound formed by chemical bonding of a reactive functional group on one molecule with a suitable reactive functional group on another molecule.

[0047] As used herein, the term "derivatized antibody" refers to an antibody that has been modified by chemical reactions to cleave residues or add chemical moieties not native to the isolated antibody. Derivatized antibodies can be obtained by glycosylation, acetylation, pegylation, phosphorylation, amidation, derivatization by addition of known chemical protecting / blocking groups, proteolytic cleavage, or conjugation to a cellular ligand or other protein. Any of a variety of chemical modifications can be performed using established procedures by known techniques, including, but not limited to, specific chemical cleavage, acetylation, formylation, tunicamycin metabolic synthesis, and the like. Additionally, derivatives can include one or more unnatural amino acid residues, for example, using amber suppression techniques (see, e.g., U.S. Pat. No. 6,964,859; this reference is incorporated herein by reference).

[0048] As used herein, the term "diabody" refers to a bivalent antibody comprising two polypeptide chains, in which each polypeptide chain is connected by a V H and V L V connected by a linker that is too short to allow intramolecular association of the domains (e.g., a linker consisting of five amino acids) H and V L domains. This structure forces each domain to pair with a complementary domain on another polypeptide chain to form a homodimeric structure. Thus, the term "triabody" refers to a trivalent antibody comprising three peptide chains, each of which contains one VH domain and one VL domain connected by a linker (e.g., a linker consisting of 1-2 amino acids) that is too short to allow intramolecular association of the VH and VL domains within the same peptide chain. To fold into their native structure, peptides constructed in this manner typically trimerize, positioning the VH and VL domains of adjacent peptide chains spatially close to each other to allow proper folding (see Holliger et al., Proc. Natl. Acad. Sci. USA 90:6444-48, 1993; this reference is incorporated herein by reference).

[0049] The term "disorder" as used herein refers to any pathological condition, disease, or pathological abnormal biological function state in a subject.In particular, the present disclosure provides ITGA11 related to disorders.Specific disorders of the present disclosure include fibrosis, inflammatory disorders, or cancer.

[0050] As used herein, the term "epitope" refers to a portion of an antigen that is recognized and bound by a polypeptide, such as an antibody, antigen-binding fragment thereof, single-chain polypeptide, or construct as described herein. With respect to a protein antigen (such as ITGA11), an epitope may be a continuous epitope, which is a single, uninterrupted segment of one or more amino acids covalently linked to each other by peptide bonds, where all of the constituent amino acids bind the polypeptide (e.g., an antibody, antigen-binding fragment thereof, single-chain polypeptide, or construct thereof). A continuous epitope may consist of, for example, 1, 5, 10, 15, 20, or more amino acids within the antigen. In some embodiments, an epitope may be a discontinuous epitope, which comprises two or more segments of amino acids, each separated from each other in the amino acid sequence of the antigen by one or more intervening amino acid residues. A non-contiguous epitope may consist of, for example, a segment of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more such amino acid residues. Despite this separation by intervening amino acids, the segments comprising a non-contiguous epitope may be located spatially close to each other, for example, in the conformation of the antigen. An epitope may be defined not only by its amino acid composition, but also by the post-translational state (e.g., phosphorylation) of the amino acids of the epitope or the bonding arrangement (e.g., cis or trans) of the peptide bond between two amino acids in the epitope.

[0051] As used herein, the term "framework region" or "FW region" includes the amino acid residues adjacent to the CDRs. FW region residues may be present, for example, in human antibodies, rodent-derived antibodies (e.g., murine antibodies), humanized antibodies, primatized antibodies, chimeric antibodies, antibody fragments (e.g., Fab fragments), single-chain antibody fragments (e.g., scFv fragments), antibody domains, and bispecific antibodies, among others.

[0052] As used herein, the term "fusion protein" refers to a protein linked to another molecule via a covalent bond. Fusion proteins can be chemically synthesized, for example, by an amide bond-forming reaction between the N-terminus of one protein and the C-terminus of another protein. Alternatively, a fusion protein comprising one protein covalently linked to another protein can be recombinantly expressed in a cell (e.g., a eukaryotic or prokaryotic cell) by, for example, expression of a polynucleotide encoding the fusion protein from a vector or the cell's genome. A fusion protein can comprise one protein covalently linked to a linker, which is in turn covalently linked to another molecule. Examples of linkers that can be used to form fusion proteins include peptide-containing linkers, such as those containing natural or unnatural amino acids. In some embodiments, it may be desirable to include D-amino acids in the linker because these residues are not present in native proteins and are therefore more resistant to degradation by endogenous proteases. Linkers can be prepared using a variety of strategies well known in the art and, depending on the reactive moiety of the linker, can be cleaved by enzymatic hydrolysis, photolysis, hydrolysis under acidic conditions, hydrolysis under alkaline conditions, oxidation, disulfide reduction, nucleophilic cleavage, or organometallic cleavage (Leriche et al., Bioorg. Med. Chem., 20:571-582, 2012).

[0053] As used herein, the term "heterospecific antibody" refers to a monoclonal antibody, preferably a human or humanized antibody, that has binding specificities for at least two different antigens. Traditionally, recombinant production of heterospecific antibodies is based on the coexpression of two immunoglobulin heavy / light chain pairs, where the two heavy chains have different specificities (Milstein et al., Nature 305:537, 1983). Similar methods are described, for example, in WO 93 / 08829, U.S. Patent Nos. 6,210,668; 6,193,967; 6,132,992; 6,106,833; 6,060,285; 6,037,453; 6,010,902; 5,989,530; 5,959,084; 5,959,083; and 5,989,530. 32,448; 5,833,985; 5,821,333; 5,807,706; 5,643,759; 5,601,819; 5,582,996, 5,496,549; 4,676,980; WO91 / 00360, WO92 / 00373; EP03089; Traunecker et al., EMBO J.10:3655 (1991); Suresh et al., Methods in Enzymology 121:210 (1986); these references are incorporated herein by reference. Heterospecific antibodies may contain Fc mutations that enhance correct chain association in multispecific antibodies, as described by Klein et al., mAbs 4(6):653-663, 2012; this reference is incorporated herein by reference.

[0054] As used herein, the term "human antibody" refers to an antibody in which substantially all portions of the protein (e.g., CDRs, framework, C L , C H Domain (e.g., C H 1. C H 2. C H 3), Hinge, (V L , V H) refers to antibodies that are substantially non-immunogenic in humans and contain only minor sequence changes or variations. Human antibodies can be produced in human cells (e.g., by recombinant expression) or in non-human animals or prokaryotic or eukaryotic cells capable of expressing functionally rearranged human immunoglobulin (e.g., heavy and / or light chain) genes. Furthermore, when a human antibody is a single-chain antibody, it can contain a linker peptide not found in naturally occurring human antibodies. For example, an Fv can contain a linker peptide, such as two to about eight glycine or other amino acid residues, connecting the heavy chain variable region and the light chain variable region. Such a linker peptide is considered to be of human origin. Human antibodies can be produced by a variety of known methods, including phage display using antibody libraries derived from human immunoglobulin sequences. See U.S. Patent Nos. 4,444,887 and 4,716,111; and International Patent Publications WO1998 / 46645; WO1998 / 50433; WO1998 / 24893; WO1998 / 16654; WO1996 / 34096; WO1996 / 33735; and WO1991 / 10741; these references are incorporated herein by reference. Human antibodies can also be produced using transgenic mice which are incapable of expressing functional endogenous immunoglobulins, but which can express human immunoglobulin genes. See, e.g., International Patent Publications WO98 / 24893; WO92 / 01047; WO96 / 34096; WO96 / 33735; U.S. Patent Nos. 5,413,923; 5,625,126; 5,633,425; 5,569,825; 5,661,016; 5,545,806; 5,814,318; 5,885,793; 5,916,771; and 5,939,598; these references are incorporated herein by reference.

[0055] As used herein, the term "humanized" antibody refers to forms of non-human (e.g., murine) antibodies that are chimeric immunoglobulins, immunoglobulin chains, or fragments thereof (such as Fv, Fab, Fab', F(ab')2, or other target-binding subdomains of antibodies) that contain minimal sequence derived from non-human immunoglobulin. Generally, humanized antibodies contain substantially all of at least one variable domain, typically two variable domains, in which all or substantially all of the CDR regions correspond to non-human immunoglobulin CDR regions. All or substantially all of the FR regions may also be human immunoglobulin sequences. Humanized antibodies may also contain at least a portion of an immunoglobulin constant region (Fc), typically of a human immunoglobulin consensus sequence. Methods for humanizing antibodies are known in the art. See, for example, Riechmann et al., Nature 332:323-7, 1988; U.S. Patent Nos. 5,530,101; 5,585,089; 5,693,761; 5,693,762; and 6,180,370; Queen et al.; EP 239400; International Patent Publication WO 91 / 09967; U.S. Patent No. 5,225,539; EP 592106; and EP 519596; these references are incorporated herein by reference.

[0056] The term "hydrophobic side chain" as used herein refers to an amino acid side chain that exhibits relatively low solubility in water, for example, due to the steric or electronic properties of the chemical moiety present in the side chain. Examples of amino acids that contain hydrophobic side chains include amino acids that contain unsaturated aliphatic hydrocarbons (such as alanine, valine, leucine, isoleucine, proline, and methionine) and amino acids that contain aromatic ring systems that are electrostatically neutral at physiological pH (such as tryptophan, phenylalanine, and tyrosine).

[0057] The term "monoclonal antibody," as used herein, refers to an antibody that is derived from a single clone, including any eukaryotic, prokaryotic, or phage clone, and not the method by which it is produced.

[0058] As used herein, the term "non-native constant region" refers to an antibody constant region that is derived from a source other than the antibody variable region or is a synthetic polypeptide produced by humans and has an amino acid sequence different from the sequence of a naturally occurring antibody constant region. For example, an antibody comprising a non-native constant region may have a variable region derived from a non-human source (e.g., mouse, rat, or rabbit) and a constant region derived from a human source (e.g., a human antibody constant region), or a constant region derived from another primate, pig, goat, rabbit, hamster, cat, dog, guinea pig, member of the bovine family (such as cow, bison, buffalo, elk, and yak, among others), cow, sheep, horse, or bison.

[0059] As used herein, the term "percent (%) sequence identity" refers to the percentage of amino acid (or nucleic acid) residues in a candidate sequence that are identical to those in a reference sequence, after aligning the sequences and, if necessary, introducing gaps to achieve the maximum percent sequence identity (e.g., gaps can be introduced into one or both of the candidate and reference sequences for optimal alignment, and non-homologous sequences can be ignored for comparison purposes). Alignment for purposes of determining percent sequence identity can be achieved in a variety of ways within the skill of those in the art, for example, using publicly available computer software (e.g., BLAST, ALIGN, or Megalign (DNASTAR) software). Those skilled in the art can determine appropriate parameters for assessing alignment, including any algorithms required to achieve maximum alignment over the entire length of the sequences being compared. For example, an aligned reference sequence for comparison with a candidate sequence may indicate that the candidate sequence exhibits 50% to 100% sequence identity over the entire length of the candidate sequence or over a selected portion of consecutive amino acid (or nucleic acid) residues of the candidate sequence. The length of the candidate sequence aligned for comparison purposes can be, for example, at least 30% (e.g., 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100%) of the length of the reference sequence. When a position in the candidate sequence is occupied by the same amino acid residue as the corresponding position in the reference sequence, then the molecules are identical at that position.

[0060] The term "primatized antibody" refers to an antibody that comprises a framework region of a primate-derived antibody and other regions, such as CDRs and / or constant regions, derived from an antibody of non-primate origin. Methods for producing primatized antibodies are known in the art. See, for example, U.S. Patent Nos. 5,658,570; 5,681,722; and 5,693,780; these references are incorporated herein by reference. For example, the primatized antibody or antigen-binding fragment thereof described herein can be produced by inserting the CDRs of a non-primate antibody or antigen-binding fragment thereof into an antibody or antigen-binding fragment thereof that comprises one or more framework regions of a primate.

[0061] As used herein, the term "operably linked" in reference to polynucleotide fragments is intended to mean that two polynucleotide fragments are joined such that the amino acid sequences encoded by the two polynucleotide fragments remain in-frame.

[0062] As used herein, the term "pharmacokinetic profile" refers to the absorption, distribution, metabolism, and clearance of a drug over time following administration of the drug to a patient.

[0063] As used herein, the term "regulatory sequence" includes promoters, enhancers, and other expression control elements (e.g., polyadenylation signals) that control the transcription or translation of antibody chain genes. Such regulatory sequences are described, for example, in Goeddel, Gene Expression Technology: Methods in Enzymology 185 (Academic Press, San Diego, CA, USA, 1990); this reference is incorporated herein by reference.

[0064] As used herein, the term "scFv" refers to a single-chain Fv antibody in which the variable domains of the heavy and light chains from an antibody are joined to form a single chain. An scFv fragment comprises a single polypeptide chain comprising the variable region of the antibody light chain (VL) (e.g., CDR-L1, CDR-L2, and / or CDR-L3) and the variable region of the antibody heavy chain (VH) (e.g., CDR-H1, CDR-H2, and / or CDR-H3), separated by a linker. The linker connecting the VL and VH regions of the scFv fragment can be a peptide linker composed of proteogenic amino acids. Alternative linkers may be used to increase the resistance of the scFv fragment to proteolysis (e.g., linkers comprising D-amino acids), to increase the solubility of the scFv fragment (e.g., hydrophilic linkers such as polyethylene glycol-containing linkers or polypeptides containing repeats of glycine and serine residues), to improve the biophysical stability of the molecule (e.g., linkers containing cysteine ​​residues that form intramolecular or intermolecular disulfide bonds), or to reduce the immunogenicity of the scFv fragment (e.g., linkers containing glycosylation sites). scFv molecules are known in the art and are described, for example, in U.S. Patent No. 5,892,019; Flo et al. (Gene 77:51, 1989); Bird et al. (Science 242:423, 1988); Pantoliano et al. (Biochemistry 30:10117, 1991); Milenic et al. (Cancer Research 51:6363, 1991); and Takkinen et al. (Protein Engineering 4:837, 1991). The VL and VH domains of scFv molecules can be derived from one or more antibody molecules. Those skilled in the art will understand that the variable regions of the scFv molecules described herein can be modified differently in the amino acid sequence of the antibody molecule from which they are derived. For example, in one embodiment, nucleotide or amino acid substitutions resulting in conservative substitutions or changes in amino acid residues can be made (e.g., in CDR and / or framework residues).Alternatively, or in addition, mutations are made to CDR amino acid residues to optimize antigen binding using art-recognized techniques. scFv fragments are described, for example, in WO2011 / 084714; this reference is incorporated herein by reference.

[0065] As used herein, the phrase "specifically binds" refers to a binding reaction that determines, for example, the presence of an antigen in a heterogeneous population of proteins and other biomolecules that are recognized with specificity by an antibody or antigen-binding fragment thereof. An antibody or antigen-binding fragment thereof that specifically binds to an antigen has a K of less than 100 nM. D For example, an antibody or antigen-binding fragment that specifically binds to an antigen has a K of at most 100 nM. D (e.g., 1 pM to 100 nM). An antibody or antigen-binding fragment thereof that does not exhibit specific binding to a particular antigen or its epitope will bind to the antigen with a K of greater than 100 nM for that particular antigen or its epitope. D (e.g., greater than 500 nM, 1 μM, 100 μM, 500 μM, or 1 mM). A variety of immunoassay formats may be used to select antibodies specifically immunoreactive with a particular protein or carbohydrate. For example, solid-phase ELISA immunoassays are routinely used to select antibodies specifically immunoreactive with a protein or carbohydrate. For a description of immunoassay formats and conditions that can be used to determine specific immunoreactivity, see Harlow & Lane, Antibodies, A Laboratory Manual, Cold Spring Harbor Press, New York (1988); and Harlow & Lane, Using Antibodies, A Laboratory Manual, Cold Spring Harbor Press, New York (1999).

[0066] As used herein, the terms "subject" and "patient" refer to an organism receiving treatment for a particular disease or condition as described herein. Examples of subjects and patients include mammals, such as humans, primates, pigs, goats, rabbits, hamsters, cats, dogs, guinea pigs, members of the bovine family (especially cows, bison, buffalo, elk, and yaks), cattle, sheep, horses, and bison, that are being treated for a disease or condition, among others.

[0067] As used herein, the term "transformation" refers to any of a variety of techniques commonly used for the introduction of foreign DNA into prokaryotic or eukaryotic host cells (e.g., electroporation, lipofection, calcium phosphate precipitation, DEAE-dextran mediated transformation, etc.).

[0068] As used herein, the term "treat" or "treatment" refers to a therapeutic procedure whose purpose is to prevent or slow (alleviate) an undesirable physiological change or disorder, such as the progression of fibrosis, an inflammatory disorder, or cancer described herein. Beneficial or desired clinical results include, but are not limited to, alleviation of symptoms, reduction in the extent of disease, stabilized (i.e., not worsening) disease state, delay or slowing of disease progression, improvement or alleviation of disease symptoms, and remission (whether partial or complete), whether detectable or undetectable. Subjects in need of treatment include those already with a condition or disorder, as well as those susceptible to the condition or disorder, or those in whom the condition or disorder is to be prevented.

[0069] The term "variable region CDR" as used herein includes amino acids in CDRs or complementarity determining regions identified using sequence or structure-based methods. As used herein, the term "CDR" or "complementarity determining region" refers to the non-contiguous antigen binding sites found in the variable regions of both heavy and light chain polypeptides. These specific regions are described by Kabat et al., J. Biol. Chem. 252:6609-6616, 1977; and Kabat et al., Sequences of Proteins of Immunological Interest, 5th Edition, U.S. Department of Health and Human Services, NIH Publication No. 91-3242, 1991; Chothia et al., J. Mol. Biol. 196:901-917, 1987; and MacCallum et al., J. Mol. Biol. 262:732-745, 1996, and the definitions herein include overlapping or subsets of amino acid residues when compared with each other. The term "CDR" may be, for example, the CDR defined by Kabat based on sequence comparison.

[0070] As used herein, the term "vector" includes nucleic acid vectors, such as DNA vectors (such as plasmids), RNA vectors, viruses, or other suitable replicons (e.g., viral vectors). Various vectors have been developed for the delivery of polynucleotides encoding foreign proteins into prokaryotic or eukaryotic cells. Examples of such expression vectors are disclosed, for example, in WO 1994 / 11026; this reference is incorporated herein by reference. The expression vectors described herein contain polynucleotide sequences and additional sequence elements used, for example, for protein expression and / or integration of these polynucleotide sequences into the genome of mammalian cells. Particular vectors that can be used for the expression of the antibodies and antibody fragments described herein include plasmids containing regulatory sequences, such as promoter and enhancer regions, that direct gene transcription. Other useful vectors for the expression of antibodies and antibody fragments contain polynucleotide sequences that increase the translation rate of these genes or improve the stability or nuclear export of mRNA resulting from gene transcription. These sequence elements include, for example, 5' and 3' untranslated regions, an internal ribosome entry site (IRES), and a polyA signal site for directing efficient transcription of the gene carried by the expression vector. The expression vectors described herein may also contain a polynucleotide encoding a marker for selection of cells containing such a vector. Examples of suitable markers include genes encoding resistance to antibiotics such as ampicillin, chloramphenicol, kanamycin, or nourseothricin.

[0071] As used herein, the term "VH" refers to the variable region of an immunoglobulin heavy chain of an antibody, including the heavy chain of an Fv, scFv, or Fab. Reference to "VL" refers to the variable region of an immunoglobulin light chain, including the light chain of an Fv, scFv, dsFv, or Fab. Antibodies (Ab) and immunoglobulins (Ig) are glycoproteins with the same structural characteristics. While antibodies exhibit binding specificity to a specific target, immunoglobulins include both antibodies and other antibody-like molecules that lack target specificity. Natural antibodies and immunoglobulins are typically heterotetrameric glycoproteins of approximately 150,000 daltons, composed of two identical light chains (L) and two identical heavy chains (H). Each heavy chain of a natural antibody has a variable domain (VH) at its amino terminus, followed by multiple constant domains. Each light chain of a natural antibody has a variable domain at its amino terminus (VL) and a constant domain at its carboxy terminus. [Brief explanation of the drawings]

[0072]

Figure 1

Figure 2

Figure 3

[0073] The present disclosure provides an antibody or antigen-binding fragment thereof that specifically binds to integrin alpha 11 (ITGA11). The present disclosure also provides a method for treating an ITGA11-related disorder in a subject, such as fibrosis, an inflammatory disorder, or cancer. The present disclosure also includes related pharmaceutical compositions, polynucleotides, vectors, host cells, production methods, therapeutic methods, diagnostic methods, and kits.

[0074] [I] Anti-ITGA11 antibody or antigen-binding fragment thereof The present disclosure provides antibodies or antigen-binding fragments thereof that specifically bind to integrin chain alpha 11 (ITGA11). In some embodiments, the antibodies or antigen-binding fragments disclosed herein specifically bind to human ITGA11, cynomolgus monkey ITGA11, and / or mouse ITGA11. In some embodiments, the antibodies or antigen-binding fragments disclosed herein specifically bind to ITGA11 (alpha 11beta 1 or ITGA11B1) heterodimerized with integrin chain beta 1 (ITGB1) (e.g., human ITGA11B1, cynomolgus monkey ITGA11B1, or mouse ITGA11B1). In some embodiments, the antibodies or antigen-binding fragments disclosed herein specifically bind to human ITGA11 heterodimerized with human ITGB1, cynomolgus monkey ITGB1, or mouse ITGB1.

[0075] In some embodiments, the antibodies or antigen-binding fragments disclosed herein specifically bind to ITGA11 (e.g., human ITGA11) rather than ITGA10 (e.g., human ITGA10) or ITGA2 (e.g., human ITGA2). In some embodiments, the antibodies or antigen-binding fragments disclosed herein specifically bind to ITGA11B1 (e.g., human ITGA11B1) rather than ITGA10B1 (e.g., human ITGA10B1) or ITGA2B1 (e.g., human ITGA2B1).

[0076] In some embodiments, the antibodies or antigen-binding fragments thereof disclosed herein are monovalent bispecific antibodies.

[0077] In some embodiments, the antibodies or antigen-binding fragments thereof disclosed herein selectively bind ITGA11 and ITGA10 compared to ITGA2 or ITGB1.

[0078] In particular, the present disclosure features anti-ITGA11 antibodies or antigen-binding fragments described by any of the CDR, heavy chain (VH), and / or light chain (VL) sequences provided below. The present disclosure specifically contemplates antibodies having any combination of the disclosed CDRs (e.g., any combination of CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, and CDR-L3 disclosed herein). The present disclosure also specifically contemplates pairing of a heavy chain (VH) comprising any of the heavy chain variable regions described herein with a light chain (VL) comprising any of the light chain variable regions disclosed herein.

[0079] mAb FIB-918-1 mAb FIB-918-1 comprises complementarity determining region (CDR) heavy chain 1 (CDR-H1) comprising the amino acid sequence of GFTFSSYA (SEQ ID NO: 1); CDR heavy chain 2 (CDR-H2) comprising the amino acid sequence of ISGSGGST (SEQ ID NO: 2); and CDR heavy chain 3 (CDR-H3) comprising the amino acid sequence of AKDLDWSGHDAFDI (SEQ ID NO: 3). mAb FIB-918-1: EVQLVESGGGLVQPGGSLRLSCAAS GFTFSSYA MSWVRQAPGKGLEWVSA ISGSGGST YYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYC AKDLDWSGHDAFDI It comprises a heavy chain variable domain having the amino acid sequence of WGQGTTVTVSS (SEQ ID NO: 7; CDRs are underlined). mAb FIB-918-1 comprises complementarity determining region (CDR) light chain 1 (CDR-L1) comprising the amino acid sequence of QSISSY (SEQ ID NO: 4); CDR light chain 2 (CDR-L2) comprising the amino acid sequence of AAS (SEQ ID NO: 5); and CDR light chain 3 (CDR-L3) comprising the amino acid sequence of QQTYSTPLT (SEQ ID NO: 6). mAb FIB-918-1 is DIQMTQSPSSLSASVGDRVTITCRAS QSISSY LNWYQQKPGKAPKLLIY AAS SLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYC QQTYSTPLT It comprises a light chain variable domain having the amino acid sequence of FGGGTKVEIKR (SEQ ID NO: 8; CDRs are underlined).

[0080] mAb FIB-918-2 mAb FIB-918-2 comprises a CDR-H1 having the amino acid sequence of GFTFSSYA (SEQ ID NO: 1); a CDR-H2 having the amino acid sequence of ISGSGGST (SEQ ID NO: 2); and a CDR-H3 having the amino acid sequence of AKDPRGSGRDDAFDI (SEQ ID NO: 20). [ka] and a heavy chain variable domain having the amino acid sequence: mAb FIB-918-2 comprises CDR-L1 having the amino acid sequence of QTIGSY (SEQ ID NO: 21); CDR-L2 having the amino acid sequence of GAS (SEQ ID NO: 22); and CDR-L3 having the amino acid sequence of QQSYSTPFT (SEQ ID NO: 11). [ka] and a light chain variable domain having the amino acid sequence:

[0081] mAb FIB-918-3 mAb FIB-918-3 It includes CDR-H1 containing the amino acid sequence of GFTFSSYS (SEQ ID NO: 14); CDR-H2 containing the amino acid sequence of ISSSSTI (SEQ ID NO: 15); and CDR-H3 containing the amino acid sequence of ARGPDLSDYFDY (SEQ ID NO: 16). mAb FIB-918-3 EVQLVESGGGLVQPGGSLRLSCAAS GFTFSSYS MNWVRQAPGKGLEWVSY ISSSSSTI YYADSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYC ARGPDLSDYFDY WGRGTLVTVSS (SEQ ID NO: 18; CDRs are underlined). mAb FIB-918-3 contains CDR-L1 comprising the amino acid sequence of QSISSY (SEQ ID NO: 4); CDR-L2 comprising the amino acid sequence of AAS (SEQ ID NO: 5); and CDR-L3 comprising the amino acid sequence of QQSYSTPLT (SEQ ID NO: 17). Includes: mAb FIB-918-3 is DIQLTQSPSSLSASVGDRVTITCRAS QSISSY LNWYQQKPGKAPKLLIY AAS SLQSGVPSRFSGSGSGTDFTLTISSLQPEDIATYYC QQSYSTPLT It comprises a light chain variable domain having the amino acid sequence of FGGGTKLEIKR (SEQ ID NO: 19; CDRs are underlined).

[0082] mAb FIB-918-4 mAb FIB-918-4 comprises CDR-H1 having the amino acid sequence of GYTFTDYY (SEQ ID NO: 28); CDR-H2 having the amino acid sequence of FDPEDGET (SEQ ID NO: 29); and CDR-H3 having the amino acid sequence of ATLDYRGVVYFDY (SEQ ID NO: 30). mAb FIB-918-4: QVQLVQSGAEVKKPGATVKISCKVS GYTFTDYY MHWVRQAPGKGLEWMGG FDPEDGET IYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYC ATLDYRGVVYFDY It comprises a heavy chain variable domain having the amino acid sequence of WGQGTLVTVSS (SEQ ID NO: 34; CDRs are underlined). mAb FIB-918-4 comprises CDR-L1 having the amino acid sequence of SGSIASNY (SEQ ID NO: 31); CDR-L2 having the amino acid sequence of EDK (SEQ ID NO: 32); and CDR-L3 having the amino acid sequence of QSYDSSNHWV (SEQ ID NO: 33). mAb FIB-918-4: NFMLTQPHSVSDSPGKTVTISCTGS SGSIASNY VQWYQQRPGSAPTTVIY EDK RRPSGVPDRFIGSIDSSSNSASLTISGLRTEDEADYYC QSYDSSNHWV It comprises a light chain variable domain having the amino acid sequence of FGGGTQLTVLG (SEQ ID NO: 35; CDRs are underlined).

[0083] mAb FIB-918-5 mAb FIB-918-5 comprises a CDR-H1 having the amino acid sequence of GFTFSSYA (SEQ ID NO: 1); a CDR-H2 having the amino acid sequence of ISGSGGST (SEQ ID NO: 2); and a CDR-H3 having the amino acid sequence of AKDPTTMTTDAFDI (SEQ ID NO: 25). mAb FIB-918-5: EVQLVESGGGLVQPGGSLRLSCAAS GFTFSSYA MSWVRQAPGKGLEWVSA ISGSGGST YYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYC AKDPTTMTTDAFDI It comprises a heavy chain variable domain having the amino acid sequence of WGQGTMVTVSS (SEQ ID NO: 26; CDRs are underlined). mAb FIB-918-5 comprises CDR-L1 having the amino acid sequence of QSISSY (SEQ ID NO: 4); CDR-L2 having the amino acid sequence of AAS (SEQ ID NO: 5); and CDR-L3 having the amino acid sequence of QQSYSTPFT (SEQ ID NO: 11). mAb FIB-918-5 is DIQMTQSPSSLSASVGDRVTITCRAS QSISSY LNWYQQKPGKAPKLLIY AAS SLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYC QQSYSTPFTIt comprises a light chain variable domain having the amino acid sequence of FGPGTKLEIKR (SEQ ID NO: 27; CDRs are underlined).

[0084] mAb FIB-918-6 mAb FIB-918-6 comprises a CDR-H1 having the amino acid sequence of GFTFSNAW (SEQ ID NO: 9); a CDR-H2 having the amino acid sequence of ISGSGGST (SEQ ID NO: 2); and a CDR-H3 having the amino acid sequence of ARDRGYSYSETSNDAFDI (SEQ ID NO: 10). mAb FIB-918-6: EVQLLESGGDLVKAGGSLRLSCAAS GFTFSNAW MSWVRQAPGKGLEWVSG ISGSGGST YYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYC ARDRGYSYSETSNDAFDI It comprises a heavy chain variable domain having the amino acid sequence of WGRGTLVTVSS (SEQ ID NO: 12; CDRs are underlined). mAb FIB-918-6 contains CDR-L1 comprising the amino acid sequence of QSISSY (SEQ ID NO: 4); CDR-L2 comprising the amino acid sequence of AAS (SEQ ID NO: 5); and CDR-L3 comprising the amino acid sequence of QQSYSTPFT (SEQ ID NO: 11). Includes: mAb FIB-918-6 is DIQMTQSPSSLSASVGDRVTITCRAS QSISSY LNWYQQKPGKAPKLLIY AAS SLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQ QSYSTPFT It comprises a light chain variable domain having the amino acid sequence of FGPGTKLEIKR (SEQ ID NO: 13; CDRs are underlined).

[0085] mAb FIB-918-7 mAb FIB-918-7 comprises a CDR-H1 having the amino acid sequence of GFMFDTHA (SEQ ID NO: 46); a CDR-H2 having the amino acid sequence of ISGSGGSI (SEQ ID NO: 47); and a CDR-H3 having the amino acid sequence of ARSGETAGTDYFDY (SEQ ID NO: 48). mAb FIB-918-7: EVQLVESGGGLVQPGGSLRLSCAAS GFMFDTHA MSWVRQAPGKGLEWVSS ISGSGGSI YYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYC ARSGETAGTDYFDY It comprises a heavy chain variable domain having the amino acid sequence of WGQGTLVTVSS (SEQ ID NO: 51; CDRs are underlined). mAb FIB-918-7 comprises CDR-L1 having the amino acid sequence of QSVSSSY (SEQ ID NO: 49); CDR-L2 having the amino acid sequence of GAS (SEQ ID NO: 22); and CDR-L3 having the amino acid sequence of QQDYNSPYT (SEQ ID NO: 50). mAb FIB-918-7 is EIVMTQSPATLSLSPGERATLSCRAS QSVSSSY LSWYQQKPGQAPRLLIY GAS TRATGIPARFSGSGSGTDFTLTISSLQPEDFAVYYC QQDYNSPYT It comprises a light chain variable domain having the amino acid sequence of FGQGTKVDIKR (SEQ ID NO: 52; CDRs are underlined).

[0086] mAb FIB-918-8 mAb FIB-918-8 comprises a CDR-H1 having the amino acid sequence of GFTFSNAW (SEQ ID NO: 9); a CDR-H2 having the amino acid sequence of ISGSGGST (SEQ ID NO: 2); and a CDR-H3 having the amino acid sequence of AKQTVTSADDYFDY (SEQ ID NO: 43). mAb FIB-918-8: EVQLVESGGGVVRPGGPLRLSCAAS GFTFSNAW MSWVRQAPGKGLEWVSA ISGSGGST YYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAIYYC AKQTVTSADDYFDY It comprises a heavy chain variable domain having the amino acid sequence of WGQGTLVTVSS (SEQ ID NO: 44; CDRs are underlined). mAb FIB-918-8 comprises CDR-L1 having the amino acid sequence of QSISSY (SEQ ID NO: 4); CDR-L2 having the amino acid sequence of AAS (SEQ ID NO: 5); and CDR-L3 having the amino acid sequence of QQSYSTPFT (SEQ ID NO: 11). mAb FIB-918-8 is DIQMTQSPSSLSASVGDRVTITCRAS QSISSY LNWYQQKPGKAPKLLIY AAS SLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYC QQSYSTPFT It comprises a light chain variable domain having the amino acid sequence of FGPGTKLEIKR (SEQ ID NO: 45; CDRs are underlined).

[0087] mAb FIB-918-9 mAb FIB-918-9 comprises a CDR-H1 having the amino acid sequence of GFTFSDYW (SEQ ID NO: 36); a CDR-H2 having the amino acid sequence of ISGSGGST (SEQ ID NO: 2); and a CDR-H3 having the amino acid sequence of AKDLLWAARDAFDI (SEQ ID NO: 37). mAb FIB-918-9: EVQLVESGGGLVQPGGSLRLSCLAS GFTFSDYW MAWVRQAPGKGLEWVSA ISGSGGST YYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYC AKDLLWAARDAFDI It comprises a heavy chain variable domain having the amino acid sequence of WGQGTLVTVSS (SEQ ID NO: 38; CDRs are underlined). mAb FIB-918-9 comprises CDR-L1 having the amino acid sequence of QSISSY (SEQ ID NO: 4); CDR-L2 having the amino acid sequence of AAS (SEQ ID NO: 5); and CDR-L3 having the amino acid sequence of QQSYSTPFT (SEQ ID NO: 11). mAb FIB-918-9 is DIQMTQSPSSLSASVGDRVTITCRAS QSISSY LNWYQQKPGKAPKLLIY AAS SLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYC QQSYSTPFTIt comprises a light chain variable domain having the amino acid sequence of FGPGTKLEIKR (SEQ ID NO: 39; CDRs are underlined).

[0088] mAb FIB-918-10 mAb FIB-918-10 comprises a CDR-H1 having the amino acid sequence of GFTFSSYA (SEQ ID NO: 1); a CDR-H2 having the amino acid sequence of ISGSGGST (SEQ ID NO: 2); and a CDR-H3 having the amino acid sequence of AKDLLWAARDAFDI (SEQ ID NO: 37). mAb FIB-918-10: QVQLVESGGGLVQPGGSLRLSCAAS GFTFSSYA MSWVRQAPGKGLEWVSA ISGSGGST YYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYC AKDLLWAARDAFDI It comprises a heavy chain variable domain having the amino acid sequence of WGQGTLVTVSS (SEQ ID NO: 41; CDRs are underlined). mAb FIB-918-10 comprises CDR-L1 having the amino acid sequence of QGINDF (SEQ ID NO: 40); CDR-L2 having the amino acid sequence of AAS (SEQ ID NO: 5); and CDR-L3 having the amino acid sequence of QQSYSTPLT (SEQ ID NO: 17). mAb FIB-918-10 is AIQLTQSPSTLSASVGDRVTITCRAS QGINDF LAWYQQKPGKAPKLLIY AAS SLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYC QQSYSTPLT It comprises a light chain variable domain having the amino acid sequence of FGGGTKVEIKR (SEQ ID NO: 42; CDRs are underlined).

[0089] Light chain variable domain sequence The present disclosure provides an antibody or antigen-binding fragment thereof comprising a light chain variable domain comprising an amino acid sequence having at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of any one of SEQ ID NOs: 8, 13, 19, 24, 35, 39, 42, 45, and 52. SEQ ID NO:8 DIQMTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQTYSTPLTFGGGTKVEIKR SEQ ID NO:13 DIQMTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPFTFGPGTKLEIKR SEQ ID NO:19 DIQLTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDIATYYCQQSYSTPLTFGGGTKLEIKR SEQ ID NO:24 NIQMTQSPSSLSASVGDRVTITCRASQTIGSYLNWYQQKPGTAPKLLIYGASTSHTWVPSRFTGGGSGTEFTLTISSLQSEDFATYYCQQSYSTPFTFGGGTKLEIKR SEQ ID NO:27 DIQMTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPFTFGPGTKLEIKR SEQ ID NO:35 NFMLTQPHSVSDSPGKTVTISCTGSSGSIASNYVQWYQQRPGSAPTTVIYEDKRRPSGVPDR Figure SIDSSSNSASLTISGLRTEDEADYYCQSYDSSNHWVFGGGTQLTVLG Accession number: 39 DIQMTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPFTFGPGTKLEIKR Accession number: 42 AIQLTQSPSTLSASVGDRVTITCRASQGINDFLAWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPLTFGGGTKVEIKR Accession number: 45 DIQMTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPFTFGPGTKLEIKR Accession number: 52 EIVMTQSPATLSLSPGERATLSCRASQSVSSSYLSWYQQKPGQAPRLLIYGASTRATGIPARFSGSGSGTDFTLTISSLQPEDFAVYYCQQDYNSPYTFGQGTKVDIKR

[0090] Heavy chain variable domain sequence The present disclosure provides an antibody or an antigen-binding fragment thereof comprising a heavy chain variable domain comprising an amino acid sequence having at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to any one of the amino acid sequences of SEQ ID NO: 7, 12, 18, 23, 26, 34, 38, 41, 44, and 51. SEQ ID NO: 7 EVQLVESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAISGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKDLDWSGHDAFDIWGQGTTVTVSS SEQ ID NO:12 EVQLLESGGDLVKAGGSLRLSCAASGFTFSNAWMSWVRQAPGKGLEWVSGISGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDRGYSYSETSNDAFDIWGRGTLVTVSS SEQ ID NO:18 EVQLVESGGGLVQPGGSLRLSCAASGFTFSSYSMNWVRQAPGKGLEWVSYISSSSSTIYYADSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCARGPDLSDYFDYWGRGTLVTVSS SEQ ID NO:23 EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAISGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKDPRGGSGRDDAFDIWGQGTMVTVSS SEQ ID NO:26 EVQLVESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAISGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKDPTTMTTDAFDIWGQGTMVTVSS SEQ ID NO:34 QVQLVQSGAEVKKPGATVKISCKVSGYTFTDYYMHWVRQAPGKGLEWMGGFDPEDGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCATLDYRGVVYFDYWGQGTLVTVSS SEQ ID NO:38 EVQLVESGGGLVQPGGSLRLSCLASGFTFSDYWMAWVRQAPGKGLEWVSAISGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKDLLWAARDAFDIWGQGTLVTVSS SEQ ID NO:41 QVQLVESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAISGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKDLLWAARDAFDIWGQGTLVTVSS SEQ ID NO:44 EVQLVESGGGVVRPGGPLRLSCAASGFTFSNAWMSWVRQAPGKGLEWVSAISGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAIYYCAKQTVTSADDYFDYWGQGTLVTVSS SEQ ID NO:51 EVQLVESGGGLVQPGGSLRLSCAASGFMFDTHAMSWVRQAPGKGLEWVSSISGSGGSIYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARSGETAGTDYFDYWGQGTLVTVSS

[0091] The antibodies described herein include fully human antibodies, humanized antibodies, primatized antibodies, and chimeric antibodies. Additionally, the antibodies described herein include fully human antibodies, humanized antibodies, primatized antibodies, and chimeric antibodies that contain one or more or all of the CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, and CDR-L3 sequences described herein, in which one or more or all of the CDR sequences are identical to those of the antibodies described herein (e.g., mAbs). and FIB-918-1, FIB-918-2, FIB-918-3, FIB-918-4, FIB-918-5, FIB-918-6, FIB-918-7, FIB-918-8, FIB-918-9, or FIB-918-10) exhibiting at least 70% sequence identity (e.g., 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity) to the corresponding CDR sequences of any one of FIB-918-1, FIB-918-2, FIB-918-3, FIB-918-4, FIB-918-5, FIB-918-6, FIB-918-7, FIB-918-8, FIB-918-9, or FIB-918-10).

[0092] In some embodiments, the antibody or antigen-binding fragment is a humanized antibody or antigen-binding fragment comprising one or more or all of the CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, and CDR-L3 sequences described herein, in which one or more or all of the CDR sequences are humanized antibodies described herein (e.g., mAbs). and FIB-918-1, FIB-918-2, FIB-918-3, FIB-918-4, FIB-918-5, FIB-918-6, FIB-918-7, FIB-918-8, FIB-918-9, or FIB-918-10).

[0093] The antibodies and antibody fragments described herein further include fully human antibodies, humanized antibodies, primatized antibodies, and chimeric antibodies comprising one or more or all of the CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, and CDR-L3 sequences, one or more or all of which CDR sequences comprise one or more (e.g., up to three) amino acid substitutions (e.g., one or more conservative amino acid substitutions) relative to the corresponding CDR sequences of an antibody described herein. For example, an antibody described herein can be generated by incorporating any one or more of the CDR sequences of an antibody described herein into the framework regions (e.g., FW1, FW2, FW3, and FW4) of a human antibody.

[0094] As an example, one strategy used to design the humanized antibodies described herein is to align the sequences of the heavy and light chain variable regions of the antibodies described herein with the heavy and light chain variable regions of consensus human antibodies. Consensus human antibody heavy and light chain sequences are known in the art (see, e.g., the "VBASE" human germline sequence database; see also Kabat et al., Sequences of Proteins of Immunological Interest, 5th ed., U.S. Department of Health and Human Services, NIH Publication No. 91-3242, 1991; Tomlinson et al., J. Mol. Biol. 227:776-98, 1992; and Cox et al., Eur. J. Immunol. 24:827-836, 1994; the disclosures of these references are incorporated herein by reference). In this way, framework residues and CDRs of the variable domains can be identified by sequence alignment (see Kabat, supra). To create a humanized antibody, for example, one or more CDRs of a consensus human antibody can be replaced with the corresponding CDRs of an antibody described herein. An example of a consensus human antibody variable domain is the heavy chain variable domain described in U.S. Patent No. 6,054,297; the disclosures of these references are incorporated herein by reference. These amino acid substitutions are carried out, for example, by recombinant expression of polynucleotides encoding the heavy and light chains of the humanized antibody in host cells using methods known in the art or described herein.

[0095] Similarly, this strategy can also be used to generate primatized antibodies; for example, one or more or all of the CDRs of a primate antibody consensus sequence can be replaced with one or more or all of the CDRs of an antibody described herein, for example. Consensus primate antibody sequences are known in the art (see, e.g., U.S. Patent Nos. 5,658,570; 5,681,722; and 5,693,780; the disclosures of each of these references are incorporated herein by reference).

[0096] In some embodiments, it may be desirable to introduce certain framework residues into the heavy and / or light chain variable domains of a human antibody in addition to the CDR sequences from an antibody, such as the antibodies described herein. For example, U.S. Patent No. 6,054,297 identifies several cases in which it is advantageous to retain certain framework residues from a particular variable region of the heavy or light chain of an antibody in the resulting humanized antibody. In some embodiments, framework residues may participate in noncovalent interactions with antigens and thus contribute to the affinity of the antibody for its target antigen. In some embodiments, individual framework residues may modulate the conformation of the CDRs and thus indirectly affect antibody-antigen interactions. Certain framework residues may form an interface between the VH and VL domains and thus contribute to the overall antibody structure. In some cases, framework residues may constitute functional glycosylation sites (e.g., Asn-X-Ser / Thr), which, upon attachment of carbohydrate moieties, may determine antibody structure and antigen affinity. In such cases, it may be advantageous to retain particular framework residues of the antibodies described herein, for example, in a humanized or primatized antagonistic antibody or antigen-binding fragment thereof, as different framework residues may promote higher epitope affinity and improved biochemical activity of the antibody or antigen-binding fragment thereof.

[0097] The antibodies described herein also include antibody fragments, Fab domains, F(ab') molecules, F(ab')2 molecules, single-chain variable fragments (scFvs), tandem scFv fragments, diabodies, triabodies, dual variable region domain immunoglobulins, multispecific antibodies, bispecific antibodies, and heterospecific antibodies, comprising one or more or all of the CDRs of an antibody described herein, or one or more or all of the CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, and CDR-L3 sequences, wherein one or more or all of the CDR sequences exhibit at least 70% sequence identity (e.g., 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity) to the corresponding CDR sequences of an antibody described herein. The antibodies described herein further include fully human, humanized, primatized, and chimeric antibodies comprising one or more or all of the CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, and CDR-L3 sequences, one or more or all of which CDR sequences comprise one or more (e.g., up to three) amino acid substitutions (e.g., one or more conservative amino acid substitutions) relative to the corresponding CDR sequences of an antibody described herein. These molecules can be expressed recombinantly, for example, by incorporating polynucleotides encoding these proteins into expression vectors for transformation into eukaryotic or prokaryotic cells using techniques described herein or known in the art, or can be synthesized, for example, by solid phase peptide synthesis methods described herein or known in the art.

[0098] The polypeptides described herein further include antibody-like molecular scaffolds comprising one or more or all of the CDRs of the antibodies described herein, or one or more or all of the CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, and CDR-L3 sequences, for example, where one or more or all of the CDR sequences exhibit at least 70% sequence identity (e.g., 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity) to the corresponding CDR sequences of the antibodies described herein or contain one or more (e.g., up to three) amino acid substitutions (e.g., one or more conservative amino acid substitutions) relative to the corresponding CDR sequences of the antibodies described herein. Exemplary antibody-like molecular scaffolds include the tenth fibronectin type III domain ( 10 Examples include proteins containing Fn3. 10 The tertiary structure of the Fn3 domain is similar to that of the variable region of an IgG heavy chain, and one of skill in the art will recognize the residues of the corresponding CDR sequences of the antibodies or antigen-binding fragments thereof described herein. 10 By substituting residues in the BC, DE, and FG loops of Fn3, it is possible to graft onto the fibronectin scaffold, for example, one or more or all of the CDR sequences of an antibody or antigen-binding fragment thereof described herein, or a sequence having at least 70% sequence identity (e.g., 75%, 80%, 85%, 90%, 95%, 97%, 99%, or 100% sequence identity) to any one or more of these CDR sequences, or a sequence containing amino acid substitutions, such as conservative or non-conservative amino acid substitutions (e.g., substitutions of up to three amino acids) to one or more of these CDR sequences. This is a modified 10 This can be achieved by recombinant expression of the Fn3 domain (e.g., using the vectors and techniques described herein) as an antibody-like molecular scaffold for grafting of antibody-derived CDRs onto the BC, DE, and FG structural loops. 10Examples using Fn3 domains are reported in WO2000 / 034784, WO2009 / 142773, WO2012 / 088006, and U.S. Patent No. 8,278,419; the disclosures of each of these references are incorporated herein by reference.

[0099] [II] Nucleic acids and expression systems The antibodies or antigen-binding fragments thereof described herein can be prepared by any of a variety of established techniques. For example, the anti-ITGA11 antibodies or antigen-binding fragments thereof described herein can be prepared by recombinantly expressing immunoglobulin light and heavy chain genes in host cells. To express an antibody by recombinant technology, host cells can be transformed with one or more recombinant expression vectors carrying DNA fragments encoding the antibody immunoglobulin light and heavy chains, so that the light and heavy chains are expressed in the host cells and, optionally, secreted into the medium in which the host cells are cultured, from which the antibody can be recovered. Standard recombinant DNA techniques are used to obtain antibody heavy and light chain genes, incorporate these genes into recombinant expression vectors, and introduce the vectors into host cells, such as those described in Molecular Cloning: A Laboratory Manual, 2nd Edition (Sambrook, Fritsch, and Maniatis (eds.), Cold Spring Harbor, NY, 1989), Current Protocols in Molecular Biology (Ausubel et al., eds., Greene Publishing Associates, 1989), and U.S. Pat. No. 4,816,397; the disclosures of each of these references are incorporated herein by reference.

[0100] Vector for expressing anti-ITGA11 antibody Viral genomes provide a rich source of vectors that can be used for the efficient introduction of foreign genes into the genome of cells (e.g., eukaryotic or prokaryotic cells). Viral genomes are particularly useful vectors for gene transfer because polynucleotides contained within such genomes are typically integrated into the genome of target cells by general or specific transduction. These processes occur as part of the natural viral replication cycle and do not require added proteins or reagents to induce gene integration. Examples of viral vectors include retroviruses, adenoviruses (e.g., Ad5, Ad26, Ad34, Ad35, and Ad48), parvoviruses (e.g., adeno-associated viruses), coronaviruses, negative-strand RNA viruses (such as orthomyxoviruses (e.g., influenza virus)), rhabdoviruses (e.g., rabies and vesicular stomatitis viruses), paramyxoviruses (e.g., measles and Sendai virus), positive-strand RNA viruses (such as picornaviruses and alphaviruses), and double-stranded DNA viruses including adenoviruses, herpesviruses (e.g., herpes simplex virus types 1 and 2, Epstein-Barr virus, cytomegalovirus), and poxviruses (e.g., vaccinia, modified vaccinia Ankara (MVA), fowlpox, and canarypox). Other viruses useful for delivering polynucleotides encoding the light and heavy chains or antibody fragments of the antibodies described herein include, for example, Norwalk virus, togavirus, flavivirus, reovirus, papovavirus, hepadnavirus, and hepatitis virus. Examples of retroviruses include avian leukosis sarcoma virus, mammalian C, B, and D viruses, the HTLV-BLV complex, lentiviruses, and spumaviruses (see article Coffin, JM Retroviridae: The viruses and their replication in Fundamental Virology, 3rd ed. (BN Fields et al., eds., Lippincott-Raven Publishers, Philadelphia, 1996)).Other examples include murine leukemia viruses, murine sarcoma viruses, mouse mammary tumor viruses, bovine leukemia viruses, feline leukemia viruses, feline sarcoma viruses, avian leukemia viruses, human T-cell leukemia viruses, baboon endogenous viruses, gibbon leukemia viruses, Mason-Pfizer monkey viruses, simian immunodeficiency viruses, simian sarcoma viruses, Rous sarcoma viruses, and lentiviruses. Other examples of vectors are described, for example, in U.S. Patent No. 5,801,030 to McVey et al.; the disclosure of each of these references is incorporated herein by reference.

[0101] Genome editing technology In addition to viral vectors, various additional methods have been developed for integrating genes into the genome of target cells for polypeptide expression, for example, those encoding antibody light and heavy chains, single-chain polypeptides, single-chain variable fragments (scFvs), tandem scFvs, Fab domains, F(ab')2 domains, diabodies, and triabodies, among others, such as those described herein. One such method that can be used to integrate polynucleotides encoding antibodies or fragments thereof, such as those described herein, into prokaryotic or eukaryotic cells involves transposons. A transposon is a polynucleotide that encodes a transposase enzyme and contains a polynucleotide sequence or gene of interest flanked by cleavage sites at the 5' and 3' positions. Once the transposon is delivered into a cell, expression of the transposon gene begins, resulting in an active enzyme that cleaves the gene of interest from the transposon. This activity is mediated by site-specific recognition of the transposon's cleavage site by the transposase. In some embodiments, these cleavage sites may be terminal repeats or inverted terminal repeats. Once excised from the transposon, the gene of interest can be integrated into the genome of a prokaryotic or eukaryotic cell by transposase-catalyzed cleavage of a similar cleavage site present in the nuclear genome of the cell. This allows the gene encoding the antibody, or a fragment thereof, or a domain thereof, to be inserted into the cleaved nuclear DNA at the excision site, and subsequent ligation of a phosphodiester bond linking the gene of interest to the DNA of the genome of the prokaryotic or eukaryotic cell completes the integration process. In some embodiments, the transposon may be a retrotransposon, whereby the gene encoding the antibody is first transcribed into an RNA product and then reverse-transcribed into DNA before integration into the genome of the prokaryotic or eukaryotic cell.Examples of transposon systems include the PiggyBac transposon (described in detail in WO2010 / 085699) and the sleeping beauty transposon (described in detail in U.S. Patent No. 20050112764); the disclosures of each of these references are incorporated herein by reference.

[0102] Another useful method for integrating nucleic acid molecules encoding antibodies or fragments thereof, such as those described herein, into the genome of prokaryotic or eukaryotic cells is the cluster of regularly interspaced short palindromic repeats (CRISPR) / Cas system, which first evolved as an adaptive defense mechanism in bacteria and archaea against viral infection. The CRISPR / Cas system consists of palindromic repeat sequences within plasmid DNA and the associated Cas9 nuclease. This combination of DNA and protein directs site-specific DNA cleavage of target sequences by first integrating foreign DNA into the CRISPR locus. Polynucleotides containing these foreign sequences and the repetitive spacer elements of the CRISPR locus are then transcribed in the host cell to generate guide RNAs, which then anneal to the target sequence and localize the Cas9 nuclease to this site. In this way, highly site-specific Cas9-mediated DNA cleavage can occur in foreign polynucleotides because the interaction that brings Cas9 into close proximity with the target DNA molecule is governed by RNA:DNA hybridization. As a result, CRISPR / Cas system can be theoretically designed to cut any target DNA molecule of interest.This technology has been utilized to edit eukaryotic genomes (Hwang et al., Nat Biotech., 31:227-229, 2013), and can be used as an efficient method for site-specific editing of eukaryotic or prokaryotic genomes to cut DNA before the integration of the polynucleotide encoding antibody or its fragment described herein.The use of CRISPR / Cas to regulate gene expression is described in U.S. Patent No. 8,697,359; the disclosure of this reference is incorporated herein by reference.

[0103] Another method for site-specifically cleaving genomic DNA before the integration of a polynucleotide encoding an antibody or its fragment, such as those described herein, includes the use of zinc finger nucleases and transcription activator-like effector nucleases (TALENs).Unlike the CRISPR / Cas system, these enzymes do not contain a guided polynucleotide for localization to a specific target sequence.Target specificity is instead controlled by the DNA binding domain within these enzymes.Zinc finger nucleases and TALENs for use in genome editing applications are described in Urnov et al. (Nat.Rev.Genet., 11:636-646, 2010); and Joung et al. (Nat.Rev.Mol.Cell.Bio.14:49-55, 2013); these references are incorporated herein by reference. Further genome editing techniques that can be used to integrate the polynucleotide encoding the antibody described herein into the genome of prokaryotic or eukaryotic cells include the use of ARCUS® meganuclease, which can be rationally designed to site-specifically cleave genomic DNA.The use of these enzymes to integrate the polynucleotide encoding the antibody or fragment thereof described herein into the genome of prokaryotic or eukaryotic cells is particularly advantageous in view of the structure / activity relationship established for such enzymes.Single-chain meganucleases can therefore be modified at specific amino acid positions to create nucleases that selectively cleave DNA at desired positions.These single-chain nucleases are described in detail in, for example, U.S. Patent Nos. 8,021,867 and 8,445,251; the disclosures of each of these references are incorporated herein by reference.

[0104] Polynucleotide Sequence Elements To express an antibody or fragment thereof, such as those described herein, polynucleotides encoding the light and heavy chains, partially or completely, for example, polynucleotides encoding one or more or all of the CDR sequences of an antibody or antigen-binding fragment thereof described herein, can be inserted into an expression vector, such that the gene is operably linked to transcriptional and translational control sequences. Expression vectors and expression control sequences are selected to be compatible with the expression host cell used. Polynucleotides encoding the light and heavy chain genes of an antibody or fragment thereof can be inserted into separate vectors, or, optionally, both polynucleotides can be incorporated into the same expression vector using established techniques described herein or known in the art. In addition to polynucleotides encoding the heavy and light chains of an antibody (or polynucleotides encoding a single-chain polypeptide, antibody fragment (such as an scFv molecule), or construct described herein), the recombinant expression vectors described herein can have regulatory sequences that control the expression of the antibody chain genes in a host cell. The design of the expression vector, including the selection of regulatory sequences, can depend on factors such as the choice of host cell to be transformed or the desired protein expression level. For example, suitable regulatory sequences for mammalian host cell expression include viral elements that direct high-level protein expression in mammalian cells, such as promoters and / or enhancers from cytomegalovirus (CMV) (such as the CMV promoter / enhancer), simian virus 40 (SV40) (such as the SV40 promoter / enhancer), adenovirus (e.g., the adenovirus major late promoter (AdMLP)), and polyoma. Viral regulatory elements, and sequences thereof, are described in detail in, for example, U.S. Patent Nos. 5,168,062; 4,510,245; and 4,968,615; the disclosures of each of these references are incorporated herein by reference.

[0105] In addition to the antibody chain genes and regulatory sequences, the recombinant expression vectors described herein can carry additional sequences, such as sequences that control replication of the vector in host cells (e.g., origins of replication) and selectable marker genes. Selectable marker genes facilitate selection of host cells into which the vector has been introduced (see, e.g., U.S. Pat. Nos. 4,399,216; 4,634,665; and 5,179,017). For example, typically, selectable marker genes confer resistance to cytotoxic drugs (such as G418, puromycin, blasticidin, hygromycin, or methotrexate) on the host cells into which the vector has been introduced. Suitable selectable marker genes include the dihydrofolate reductase (DHFR) gene (for use in methotrexate selection / amplification in "DHFR" host cells) and the neo gene (for G418 selection). For expression of the light and heavy chains of the antibody or fragment thereof, the expression vector containing the polynucleotides encoding the heavy and light chains can be transfected into a host cell by standard techniques.

[0106] Host cells for expression of anti-ITGA11 antibodies or fragments thereof The antibodies or fragments thereof described herein can be expressed in either prokaryotic or eukaryotic host cells. In certain embodiments, expression of the antibodies or fragments thereof is carried out in eukaryotic cells, such as mammalian host cells, for optimal secretion of properly folded and immunologically active antibodies. Examples of mammalian host cells for expressing the recombinant antibodies or antigen-binding fragments thereof described herein include Chinese hamster ovary (CHO) cells (e.g., including the DHFR CHO cells described in Urlaub and Chasin (1980, Proc. Natl. Acad. Sci. USA 77:4216-4220) used with the DHFR selectable marker as described in Kaufman and Sharp (1982, Mol. Biol. 159:601-621)), NSO myeloma cells, COS cells, 293 cells, and SP2 / 0 cells. Additional cell types that may be useful for expressing antibodies and fragments thereof include bacterial cells, such as BL-21(DE3) E. coli cells, which can be transformed with vectors containing foreign DNA according to established protocols. Additional eukaryotic cells that may be useful for expressing antibodies include yeast cells, such as autoautotrophic strains of S. cerevisiae, which can be transformed according to established and known procedures in the art and selectively grown in a deficient medium. When a recombinant expression vector encoding an antibody gene is introduced into a mammalian host cell, the antibody is produced by culturing the host cell for a sufficient period of time to allow for expression of the antibody in the host cell or secretion of the antibody into the medium in which the host cell is grown.

[0107] Antibodies or antigen-binding fragments thereof can be recovered from the culture medium using standard protein purification methods. Host cells can also be used to produce portions of intact antibodies, such as Fab fragments or scFv molecules. Methods in which the above procedures vary according to well-known protocols established in the art are also included herein. For example, to produce antigen-binding fragments of antibodies, it may be desirable to transduce host cells with DNA encoding either the light chain or the heavy chain (but not both) of an antibody or fragment thereof described herein.

[0108] Once an antibody or fragment thereof described herein is produced by recombinant expression, it can be purified by any method known in the art, such as methods useful for the purification of immunoglobulin molecules, for example, by chromatography (e.g., ion exchange, affinity, and size fractionation column chromatography), centrifugation, fractionation by differential solubility, or any other standard technique for protein purification. Additionally, the antibodies or fragments thereof described herein can be fused to heterologous polypeptide sequences described herein or known in the art to facilitate purification or to produce therapeutic conjugates.

[0109] Once isolated, the antibodies or fragments thereof may be further purified, if desired, by, for example, high performance liquid chromatography (see, e.g., Fisher, Laboratory Techniques in Biochemistry and Molecular Biology (Work and Burdon, eds., Elsevier, 1980); this reference is incorporated herein by reference) or by gel filtration chromatography, such as on a SUPERDEX® 75 column (Pharmacia Biotech AB, Uppsala, Sweden).

[0110] [III] Preparation and purification of anti-ITGA11 antibodies or their fragments Antigens containing epitopes corresponding to portions of ITGA11 can be used to generate, for example, monoclonal, polyclonal, chimeric, humanized, or recombinant ITGA11-specific antibodies. Methods include immunological methods such as those described by Kohler and Milstein (Nature 256:495-497, 1975; and Eur. J. Immunol. 6: 511-519, 1976) and Campbell ("Monoclonal Antibody Technology, The Production and Characterization of Rodent and Human Hybridomas," Burdon et al., eds., Laboratory Techniques in Biochemistry and Molecular Biology, Vol. 13, Elsevier Science Publishers, Amsterdam, 1985), and recombinant DNA methods such as those described by Huse et al. (Science 246:1275-1281, 1989).

[0111] Briefly, an antigen may be combined with an adjuvant and administered to a host animal (e.g., rabbit, mouse, rat, goat, guinea pig, hamster, horse, sheep, and non-human primate). Such administration of the antigen may be carried out by any of a variety of methods, including, but not limited to, subcutaneous injection or intramuscular injection. Upon administration, the titer of antibodies produced in the host animal is monitored, which may be carried out by any of a variety of techniques well known in the art (e.g., periodic blood sampling), and the antiserum is isolated (e.g., by centrifugation) and then screened, for example, for the presence of antibodies having binding affinity for ITGA11. Screening for the desired antibody may be carried out by techniques including, for example, radioimmunoassay, ELISA, sandwich immunoassay, immunoradiometric assay, gel diffusion precipitation reaction, in situ immunoassay (e.g., using colloidal gold, enzyme, or radioisotope labels), Western blot, precipitation reaction, agglutination assay (e.g., gel agglutination assay or hemagglutination assay), complement fixation assay, immunofluorescence assay, protein A assay, and immunoelectrophoresis assay.

[0112] Antisera obtained from the host animal may be affinity purified to obtain the antibodies of the present disclosure. The antisera may be purified by conventional techniques, such as by introducing the antisera into a separation column. The antigen of the present disclosure may be immobilized on a column to isolate and purify the antibody. The column may then be washed to remove antibodies that do not have specificity for the antigen immobilized on the column, and the remaining antibodies are eventually eluted from the column. The isolated antibodies may then be stored using conventional practices known to those of skill in the art.

[0113] Established methods for immunizing primates are known in the art (see, e.g., WO1986 / 6004782; this reference is incorporated herein by reference). Immunization is a stable method for producing monoclonal antibodies by utilizing the antigen specificity of B lymphocytes. For example, monoclonal antibodies can be prepared by the Kohler-Millstein method (e.g., the method described in EP0110716; this reference is incorporated herein by reference), in which spleen cells from a non-human animal (e.g., a primate) are provided with antigenic peptides. Clonally expanded B lymphocytes generated by immunization can be isolated from the animal's serum and then fused with myeloma cells to form hybridomas. Hybridomas are particularly useful cells for antibody production because these immortalized cells can provide a continuous supply of antigen-specific antibodies. Antibodies from such hybridomas can then be isolated using techniques known in the art, for example, by purifying the antibodies from the cell culture medium by affinity chromatography.

[0114] Alternatively, antibody libraries (e.g., naive antibody libraries, synthetic antibody libraries, semi-synthetic antibody libraries, or combinatorial libraries) can be screened to identify antibodies. Such libraries are commercially available from several sources (e.g., Cambridge Antibody, Cambridge, UK; Genetastix Corporation, Pacific Northwest Laboratory, Richland, Washington, USA; and MorphoSys AG, Munich, Germany (e.g., HuCal GOLD)). See, for example, U.S. Patent Nos. 6,696,248; 6,706,484; 6,828,422; and 7,264,963; these references are incorporated herein by reference.

[0115] Screening of antibody libraries can be performed using one of the methods known to those skilled in the art, including, for example, phage display, selectively infective phage, polysome technology, and assay systems for enzymatic activity or protein stability. Antibodies with desired properties can be identified, for example, by sequencing the corresponding nucleic acid sequence, by amino acid sequencing, or by mass spectrometry. Optimization can be performed by replacing the small sequence with a different sequence (e.g., a random sequence) and then repeating the screening process one or more times. Antibodies can be screened, for example, for optimized affinity or specificity for the target molecule, optimized expression level, optimized stability, or optimized solubility.

[0116] The antibodies of the present disclosure recognize and specifically bind to ITGA11. In some embodiments, the Kd between the antibody and ITGA11 is, for example, at least about 10 -4 M, 10 -5 M, 10 -6 M, 10 -7 M, 10 -8 M, 10 -9 M, 10 -10 M, 10 -11 M, or 10 -12 Being or exceeding M.

[0117] [IV] Anti-ITGA11 antibody conjugate For example, it may be desirable to conjugate an antibody or fragment thereof to a second molecule to modulate the activity of the antibody in vivo or for diagnostic purposes. An antibody or antigen-binding fragment thereof can be conjugated to another molecule at either the N-terminus or C-terminus of the antibody's light or heavy chain using any one of a variety of well-established conjugation strategies well known in the art. Examples of reactive functional group pairs that can be used to covalently link an antibody or antigen-binding fragment thereof to another molecule include, without limitation, thiol pairs; carboxylic acids and amino groups; ketones and amino groups; aldehydes and amino groups; thiols and alpha, beta-unsaturated moieties (such as maleimides or dehydroalanine); thiols and alpha-haloamides; carboxylic acids and hydrazides; aldehydes and hydrazides; and ketones and hydrazides.

[0118] Antibodies or antigen-binding fragments thereof can be directly covalently linked to other molecules by chemical conjugation as described. Alternatively, fusion proteins comprising the antibodies or antigen-binding fragments thereof can be recombinantly expressed from cells (e.g., eukaryotic or prokaryotic cells). This can be achieved, for example, by incorporating a polynucleotide encoding the fusion protein into the nuclear genome of the cell (e.g., using techniques described herein or known in the art). Optionally, the antibodies and fragments thereof described herein can be linked to a second molecule by forming a covalent bond between the antibody and a linker. This linker is then conjugated to the other molecule, or the linker can be conjugated to another molecule prior to ligation to the antibody or antigen-binding fragment. Examples of linkers that can be used for conjugation include polypeptide linkers, such as those containing natural or unnatural amino acids. In some embodiments, it may be desirable to include D-amino acids in the linker because these residues do not occur in natural proteins and are therefore more resistant to degradation by endogenous proteases. Fusion proteins containing polypeptide linkers can be made using chemical synthesis techniques, such as those described herein, or by recombinant expression of a polynucleotide encoding the fusion protein in a cell (e.g., a prokaryotic or eukaryotic cell). Linkers can be prepared using a variety of strategies known in the art and, depending on the reactive elements of the linker, can be cleaved by enzymatic hydrolysis, photolysis, hydrolysis under acidic conditions, hydrolysis under alkaline conditions, oxidation, disulfide reduction, nucleophilic cleavage, or organometallic cleavage (Leriche et al., Bioorg. Med. Chem., 20:571-582, 2012).

[0119] The antibodies or antigen-binding fragments thereof described herein can be conjugated to, mixed with, or administered separately from a therapeutic agent.

[0120] Labeled anti-ITGA11 antibody or its antigen-binding fragment In some embodiments, the antibodies or antigen-binding fragments thereof described herein are conjugated to other molecules (e.g., epitope tags) for purposes of purification or detection. Examples of such molecules useful in protein purification include those that display a structural epitope that can be recognized by a second molecule. This is a common strategy used in protein purification by affinity chromatography, in which a molecule is immobilized on a solid support and exposed to a heterogeneous mixture containing a target protein conjugated to a molecule that can bind to the immobilized compound. Examples of epitope tag molecules that can be conjugated to the antibodies or antigen-binding fragments thereof described herein for molecular recognition purposes include, but are not limited to, maltose-binding protein, glutathione-S-transferase, polyhistidine tag, FLAG tag, myc tag, human influenza hemagglutinin (HA) tag, biotin, and streptavidin. Complexes containing the epitopes presented by these molecules can be recognized by complementary molecules such as maltose, glutathione, nickel-containing complexes, anti-FLAG antibodies, anti-myc antibodies, anti-HA antibodies, streptavidin, or biotin, respectively. For example, antibodies described herein or fragments thereof conjugated to epitope tags derived from complex mixtures of other proteins and biomolecules (e.g., DNA, RNA, carbohydrates, phospholipids, etc.) can be purified by treating the mixture with a solid-phase resin containing a complementary molecule that can selectively recognize and bind to the epitope tag of the antibody or fragment thereof. Examples of solid-phase resins include agarose beads, which are compatible with purification in aqueous solutions.

[0121] The antibody or antigen-binding fragment thereof can also be covalently linked to a fluorescent molecule, for example, to detect the antibody or antigen-binding fragment thereof described herein by fluorometry and / or direct visualization using fluorescence microscopy. Examples of fluorescent molecules that can be conjugated to the antibodies described herein include green fluorescent protein, cyan fluorescent protein, yellow fluorescent protein, red fluorescent protein, phycoerythrin, allophycocyanin, Hoechst, 4',6-diamidino-2-phenylindole (DAPI), propidium iodide, fluorescein, coumarin, rhodamine, tetramethylrhodamine, and cyanine. Further examples of fluorescent molecules suitable for conjugation to the antibodies described herein are well known in the art and are described in detail in, for example, U.S. Patent Nos. 7,417,131 and 7,413,874; each of these references is incorporated herein by reference.

[0122] The antibodies or antigen-binding fragments thereof described herein containing fluorescent molecules are particularly useful for monitoring the cell surface localization characteristics of antibodies and their fragments. For example, cultured mammalian cells can be exposed to the antibodies or antigen-binding fragments thereof described herein covalently conjugated to fluorescent molecules, and then analyzed using conventional fluorescence microscopy techniques known in the art. Confocal fluorescence microscopy is a particularly powerful method for determining the cell surface localization of tagged antibodies, because each side of the cell can be analyzed to distinguish antibodies or fragments thereof internalized inside the cell, for example, by receptor-mediated endocytosis, from those bound to the outer surface of the cell membrane. Furthermore, cells can be treated with antibodies conjugated to fluorescent molecules that emit visible light at specific wavelengths (e.g., fluorescein, which fluoresces at approximately 535 nm), as well as additional fluorescent molecules known to localize to specific cell surface sites and fluoresce at different wavelengths (e.g., molecules that localize to CD25 and fluoresce at approximately 599 nm). The resulting emission pattern can be visualized by confocal fluorescence microscopy, and the images from these two wavelengths can be combined to reveal information about the location of the antibody or its antigen-binding fragment on the cell surface in relation to other receptors.

[0123] Bioluminescent proteins can be incorporated into fusion proteins for detection and visualization of antibodies or fragments thereof. Bioluminescent proteins, such as luciferase and aequorin, emit light as part of a chemical reaction with a substrate (e.g., luciferin and coelenterazine). Exemplary bioluminescent proteins suitable for use as diagnostic sequences and methods for their use are described, for example, in U.S. Pat. Nos. 5,292,658, 5,670,356, 6,171,809, and 7,183,092; each of these references is incorporated herein by reference. The antibodies or antigen-binding fragments thereof described herein labeled with bioluminescent proteins are useful tools for detecting antibodies in in vitro assays. For example, the presence of an antibody conjugated to a bioluminescent protein can be detected in a complex mixture of additional proteins by separating the components of the mixture using gel electrophoresis methods known in the art (e.g., native gel analysis) and then transferring the separated proteins to a membrane for Western blotting. Detection of antibodies in a mixture of other proteins can be achieved by treating the membrane with an appropriate luciferase substrate using established protocols and then visualizing the mixture of proteins on the film.

[0124] The antibodies or antigen-binding fragments thereof described herein can also be linked to a molecule containing a radioactive nuclide, such that the antibodies or fragments thereof described herein can be detected by analyzing the radioactive emission pattern of the nuclide. Alternatively, the antibodies or fragments thereof can be directly modified by incorporating the radioactive nuclide into the antibody during preparation of the protein. 35 S), nitrogen ( 15 N), or carbon ( 13Radioactive isotopes (C) can be incorporated into the antibodies or fragments thereof described herein, for example, by culturing bacteria in medium supplemented with nutrients containing these isotopes. Optionally, tyrosine derivatives containing radioactive halogens can be incorporated into antibodies, for example, by culturing bacterial cells in medium supplemented with radiolabeled tyrosine. It has been shown that tyrosine functionalized at the phenolic C2 position with radioactive halogens is rapidly incorporated into growing polypeptide chains in vivo using endogenous translation enzymes (U.S. Pat. No. 4,925,651; this reference is incorporated herein by reference). Halogens include fluorine, chlorine, bromine, iodine, and astatine. Furthermore, antibodies can be modified after isolation and purification from cell culture by functionalizing the polypeptides described herein with radioactive isotopes. Halogens are a class of isotopes that can be readily introduced into purified proteins by aromatic substitution at tyrosine or tryptophan, for example, by reaction of one or more of these residues with electrophilic halogen species. Examples of radioactive halogen isotopes include: 18 F, 75 Br, 77 Br, 122 I, 123 I, 124 I, 125 I, 129 I, 131 I, or 211 At is one example.

[0125] Another alternative strategy for incorporating radioisotopes is the covalent attachment of a chelating group to the antibody or fragment thereof, or construct. The chelating group can be covalently linked to the antibody or fragment thereof by attachment to a reactive functional group such as a thiol, amino group, alcohol, or carboxylic acid. The chelating group can then be 125 I, 67 Ga, 111 In, 99 Tc, 169 Yb, 186 Re, 123 I, 124 I, 125 I, 131 I, 99mTc, 111 In, 64 Cu, 67 Cu, 186 Re, 188 Re, 177 Lu, 90 Y, 77 As, 72 As, 86 Y, 89 Zr, 211 At, 212 Bi, 213 Bi, or 225 It can be modified to contain any of a variety of metallic radioisotopes, including but not limited to radionuclides such as Ac.

[0126] In some embodiments, the antibodies or fragments thereof described herein are administered in combination with Gd 3+ , Fe 3+ , Mn 3+ , or Cr 2+It may be desirable to covalently conjugate the antibody with a chelating group capable of binding metal ions derived from heavy elements or rare earth ions, such as chromium(III), manganese(II), iron(II), iron(III), cobalt(II), nickel(II), copper(II), praseodymium(III), neodymium(III), samarium(III), gadolinium(III), terbium(III), dysprosium(III), holmium(III), erbium(III), and ytterbium(III). In this way, the antibody can be detected by MRI spectroscopy. For example, an antibody or fragment thereof conjugated to a chelating group bound to a paramagnetic ion can be administered to a mammalian subject (e.g., a human patient) to monitor the distribution of the antibody after administration. This can be achieved by administering the antibody to a patient via any of the administration routes described herein, such as intravenous administration, and then analyzing the location of the administered antibody by recording an MRI of the patient according to established protocols. The antibody or antigen-binding fragment thereof can also be conjugated to other molecules to improve the solubility and stability of the antibody or antigen-binding fragment thereof in aqueous solution. Examples of such molecules include PEG, PSA, bovine serum albumin (BSA), and human serum albumin (HSA), among others. For example, the antibody can be conjugated to a carbohydrate moiety to avoid detection of the antibody or its fragment by the immune system of the patient receiving treatment. This process of hyperglycosylation reduces the immunogenicity of therapeutic proteins by sterically hindering the interaction of the protein with B-cell receptors in circulation. Alternatively, the antibody or fragment thereof can be conjugated to a molecule that prevents clearance of the antibodies described herein from human serum and improves their pharmacokinetic profile. Examples of molecules that may be conjugated to or inserted within the antibodies or antigen-binding fragments thereof described herein to reduce the clearance and improve the pharmacokinetic profile of these antibodies and fragments include salvage receptor binding epitopes.These epitopes are found within the Fc region of IgG immunoglobulins and have been shown to bind Fc receptors and extend the half-life of antibodies in human serum. Insertion of salvage receptor-binding epitopes into antibodies or fragments thereof can be performed, for example, as described in U.S. Patent No. 5,739,277; this reference is incorporated herein by reference.

[0127] [V] Treatment Methods The anti-ITGA11 antibodies or antigen-binding fragments thereof described herein can be used to treat patients suffering from ITGA11-related disorders (e.g., disorders affected by ITGA11 inhibition), such as fibrosis, inflammatory disorders, or cancer. The anti-ITGA11 antibodies or antigen-binding fragments thereof can be administered to mammalian subjects, such as humans, suffering from fibrosis, inflammatory disorders, or cancer.

[0128] fibrosis Fibrosis is a normal response to various tissue injuries that can lead to organ dysfunction. Diseases characterized by pathological fibrosis and that can be treated using the methods and compositions of the present invention include liver fibrosis (e.g., fibrosis associated with cirrhosis (e.g., alcohol-induced cirrhosis, viral-induced cirrhosis, cirrhosis after hepatitis C, and primary biliary cirrhosis), schistosomiasis, cholangitis (e.g., sclerosing cholangitis), and autoimmune-induced hepatitis); kidney fibrosis (e.g., tubulointerstitial fibrosis, scleroderma, diabetic nephritis, and glomerulonephritis); skin fibrosis (e.g., scleroderma, hypertrophic and keloid scarring, nephrogenic fibrosing dermatosis, and burns); myelofibrosis; neurofibromatosis; fibromas; intestinal fibrosis, and fibrous adhesions due to surgical procedures); cardiac fibrosis (e.g., fibrosis associated with myocardial infarction); vascular fibrosis (e.g., These include, but are not limited to, fibrosis associated with arterial restenosis after angioplasty and atherosclerosis; ocular fibrosis (e.g., fibrosis associated with cataract surgery, proliferative vitreoretinopathy, and retroorbital fibrosis); myelofibrosis (e.g., idiopathic myelofibrosis and drug-induced myelofibrosis); pulmonary fibrosis (e.g., interstitial pulmonary fibrosis); glomerulonephritis; heart failure (ischemic and non-ischemic); scleroderma; excessive scar tissue after surgery or device insertion, trauma or burns, progressive kidney disease, valvular heart disease, hypertensive heart disease, articular and periarticular fibrosis, myelofibrosis, ocular / vitreous fibrosis, intestinal fibrosis and strictures, peritoneal and retroperitoneal fibrosis, pancreatic fibrosis, nephrogenic systemic fibrosis, and primary sclerosing cholangitis. Pathological matrix elaboration is also involved in fibroproliferative tumor progression and metastasis.

[0129] inflammatory disease Inflammation can be classified as either acute or chronic. Acute inflammation is the body's initial response to a harmful stimulus and is caused by increased migration of plasma and white blood cells (especially granulocytes) from the blood into the damaged tissue. A series of biochemical events expands and matures the inflammatory response, involving the local vasculature, immune system, and various cells within the damaged tissue. Prolonged inflammation, known as chronic inflammation, results in a gradual change in the types of cells present at the site of inflammation and is characterized by the simultaneous destruction and healing of tissue resulting from the inflammatory process. Diseases characterized by pathological inflammation and that can be treated using the methods and compositions of the present invention include asthma (e.g., allergic asthma, exercise-induced asthma, aspirin-sensitive / exacerbated asthma, atopic asthma, severe asthma, mild asthma, moderate-to-severe asthma, corticosteroid-naive asthma, chronic asthma, corticosteroid-resistant asthma, corticosteroid-refractory asthma, newly diagnosed and untreated asthma, smoking-induced asthma, asthma uncontrolled by corticosteroids, etc.), airway inflammation, airway hyperreactivity, airway hyperresponsiveness, and the like. hyperresponsiveness), rhinosinusitis, rhinosinusitis with polyps, nasal polyps, arthritis (e.g., osteoarthritis, rheumatoid arthritis, collagen-induced arthritis, arthritis as a result of injury, etc.), eosinophilic inflammation, mast cell-mediated inflammatory diseases, sepsis, septic shock, seronegative enthesitis / arthritis (SEA) syndrome, osteoporosis, eosinophilic esophagitis, scleroderma, dermatitis, atopic dermatitis, allergic rhinitis, bullous pemphigoid, urticaria (e.g., chronic urticaria), inflammation of cartilage, polymyalgia rheumatica, polyarteritis nodosanodossa, Wegener's granulomatosis, Behçet's disease, myositis, polymyositis, dermatomyolitis, vasculitis, arteritis, diabetic nephropathy, interstitial cystitis, graft-versus-host disease (GVHD), inflammatory conditions of the gastrointestinal tract (e.g., inflammatory bowel disease (IBD), ulcerative colitis (UC), Crohn's disease (CD)), enteritis (e.g., enteritis due to environmental insults, e.g., treatment regimens (chemotherapy, radiation) enteritis caused by or associated with gastrointestinal diseases such as infectious enteritis, ischemic enteritis, collagenous or lymphocytic colitis, necrotizing enterocolitis, chronic granulomatous disease or celiac disease, food allergies, gastritis, infectious gastritis or enterocolitis (e.g., Helicobacter pylori-infected chronic active gastritis), and other types of gastrointestinal inflammation caused by infectious agents), as well as inflammatory pulmonary conditions (e.g., chronic obstructive pulmonary disease (COPD), eosinophilic pulmonary disease (OCD), pulmonary arterial ulcers (POD), and pulmonary ulcers (POD). These include: pulmonary inflammation, infection-induced lung pathologies (including those associated with viruses (e.g., influenza, parainfluenza, rotavirus, human metapneumovirus, and respiratory syncytial virus), bacterial infections, fungal (e.g., Aspergillus), parasitic infections, or prion infections, allergen-induced lung pathologies, pollutant-induced lung pathologies (e.g., asbestosis, silicosis, or beryllium poisoning), gastric aspiration-induced lung pathologies, immune dysregulation, cystic fibrosis, etc. These include, but are not limited to, genetically predisposed inflammatory conditions, physical trauma-induced pulmonary conditions (e.g., ventilator injury), emphysema, bronchitis, sarcoidosis, histiocytosis, lymphangioleiomyomatosis, acute lung injury, acute respiratory distress syndrome, chronic lung disease, bronchopulmonary dysplasia, pneumonia (e.g., community-acquired pneumonia, hospital-acquired pneumonia, ventilator-associated pneumonia, viral pneumonia, bacterial pneumonia, and severe pneumonia), airway exacerbation, and acute respiratory distress syndrome (ARDS).

[0130] cancer Cancer is a pathological condition characterized by a cell population that shows uncontrolled cell proliferation.ITGA11 signal transduction can contribute to human carcinogenesis, including invasion and metastasis.Cancer that can be treated using the method and composition of the present invention includes but is not limited to breast cancer, colorectal cancer, liver cancer, kidney cancer, liver cancer, lung cancer, pancreatic cancer, gastrointestinal cancer, melanoma, ovarian cancer, prostate cancer, cervical cancer, bladder cancer, glioblastoma, head and neck cancer and bile duct cancer.In some embodiments, the method further comprises administering at least one additional anticancer agent to the subject.

[0131] [VI] Pharmaceutical composition Pharmaceutical compositions containing the anti-ITGA11 antibodies or antigen-binding fragments thereof described herein can be prepared using methods known in the art. In particular, anti-ITGA11 antibodies or antigen-binding fragments thereof that can be incorporated into the pharmaceutical compositions of the present disclosure include anti-ITGA11 antibodies or antigen-binding fragments thereof (such as human, humanized, or chimeric variant anti-ITGA11 antibodies described herein) that contain one or more or all of the CDR sequences of the antibodies or antigen-binding fragments thereof described herein.

[0132] The pharmaceutical compositions described herein may comprise an antibody or antigen-binding fragment thereof described herein in combination with one or more pharmaceutically acceptable excipients. For example, the pharmaceutical compositions described herein can be prepared using, for example, physiologically acceptable carriers, excipients, or stabilizers (see Remington's Pharmaceutical Sciences, 16th ed., Osol, A., ed. (1980); this reference is incorporated herein by reference) and in a desired form, for example, in the form of a lyophilized formulation or aqueous solution. The compositions can also be prepared to contain an active agent at a desired concentration. For example, the pharmaceutical compositions described herein may contain at least 10% (e.g., 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 97%, 98%, 99%, 99.5%, 99.9%, or 100%) of the active agent by weight (w / w).

[0133] Furthermore, the active agent to be incorporated into the pharmaceutical formulation may itself have a desired level of purity. For example, the antibodies or antigen-binding fragments thereof described herein may be characterized by a certain degree of purity after isolation of the antibody from cell culture medium or after chemical synthesis of a single-chain antibody fragment (e.g., scFv), for example, by established solid-phase peptide synthesis methods or native chemical ligation as described herein. The antibodies or antigen-binding fragments thereof described herein may be at least 10% pure (e.g., 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 97%, 98%, 99%, 99.5%, 99.9%, 99.99%, or 100% pure) before incorporating the antibody into a pharmaceutical composition.

[0134] Pharmaceutical compositions of the antibodies or antigen-binding fragments thereof described herein can be prepared for storage as lyophilized formulations or aqueous solutions by mixing antibodies having the desired degree of purity with optional pharmaceutically acceptable carriers, excipients, or stabilizers commonly used in the art (e.g., buffers, stabilizers, preservatives, tonicity agents, non-ionic surfactants, antioxidants, and other unclassified additives). See, for example, Remington's Pharmaceutical Sciences, 16th ed., Osol, A. (ed.) (1980); this reference is incorporated herein by reference. Such additives should be nontoxic to recipients at the dosages and concentrations used.

[0135] buffer Buffering agents help maintain pH in a range close to physiological conditions. They may be present in a concentration range of about 2 mM to about 50 mM. Suitable buffers for use with the antibodies or antigen-binding fragments thereof described herein include citrate buffers (e.g., monosodium citrate / disodium citrate mixtures, citric acid / trisodium citrate mixtures, citric acid / monosodium citrate mixtures, etc.), succinate buffers (e.g., succinic acid / monosodium succinate mixtures, succinic acid / sodium hydroxide mixtures, succinic acid / disodium succinate mixtures, etc.), tartrate buffers (e.g., tartaric acid / sodium tartrate mixtures, tartaric acid / potassium tartrate mixtures, tartaric acid / sodium hydroxide mixtures, etc.), fumarate buffers (e.g., fumaric acid / monosodium fumarate mixtures, fumaric acid / disodium fumarate mixtures, etc.), and the like. , monosodium fumarate / disodium fumarate mixtures, etc.), gluconate buffers (e.g., gluconic acid / sodium gluconate mixtures, gluconic acid / sodium hydroxide mixtures, gluconic acid / potassium gluconate mixtures, etc.), oxalate buffers (e.g., oxalic acid / sodium oxalate mixtures, oxalic acid / sodium hydroxide mixtures, oxalic acid / potassium oxalate mixtures, etc.), lactate buffers (e.g., lactic acid / sodium lactate mixtures, lactic acid / sodium hydroxide mixtures, lactic acid / potassium lactate mixtures, etc.), and acetate buffers (e.g., acetic acid / sodium acetate mixtures, acetic acid / sodium hydroxide mixtures, etc.). Additionally, phosphate buffers, histidine buffers, and trimethylamine salts such as Tris may be used.

[0136] preservatives Preservatives can be added to the compositions described herein to retard microbial growth and can be added in amounts ranging from 0.2% to 1% (w / v). Suitable preservatives for use with the antibodies or antigen-binding fragments thereof described herein include phenol, benzyl alcohol, metacresol, methylparaben, propylparaben, octadecyldimethylbenzylammonium chloride, benzalkonium halides (e.g., chloride, bromide, and iodide), hexamethonium chloride, alkylparabens (such as methyl or propylparaben), catechol, resorcinol, cyclohexanol, and 3-pentanol. Tonicity agents, also known as "stabilizers," can be added to ensure isotonicity of the liquid compositions described herein and include polyhydric sugar alcohols, such as trihydric or higher sugar alcohols (e.g., glycerin, arabitol, xylitol, sorbitol, and mannitol). Stabilizers refer to a broad category of excipients, which can range in function from bulking agents to additives that solubilize the therapeutic agent or help prevent denaturation or adhesion to the container wall.Typical stabilizers include polyhydric sugar alcohols (as listed above); amino acids (arginine, lysine, glycine, glutamine, asparagine, histidine, alanine, ornithine, L-leucine, 2-phenylalanine, glutamic acid, threonine, etc.), organic sugars or sugar alcohols (including lactose, trehalose, stachyose, mannitol, sorbitol, xylitol, ribitol, myo-inositol, galactitol, glycerol, etc.; cyclitols (such as inositol)); polyethylene glycol; amino acid polymers; sulfur-containing reducing agents (urea, , glutathione, thioctic acid, sodium thioglycolate, thioglycerol, α-monothioglycerol, and sodium thiosulfate); low molecular weight polypeptides (e.g., peptides of 10 residues or less); proteins (e.g., human serum albumin, bovine serum albumin, gelatin, or immunoglobulins); hydrophilic polymers (e.g., polyvinylpyrrolidone); monosaccharides (e.g., xylose, mannose, fructose, glucose); disaccharides (e.g., lactose, maltose, sucrose), and trisaccharides (e.g., raffinose); and polysaccharides (e.g., dextran). The stabilizer may be present in an amount ranging from 0.1 to 10,000 parts by weight of active protein.

[0137] surfactants Nonionic surfactants or surfactants (also known as "wetting agents") can be added to aid in solubilizing the therapeutic agent and to protect the therapeutic protein from shaking-induced aggregation; they also allow the formulation to be exposed to surface shear stress without denaturing the protein. Suitable nonionic surfactants include polysorbates (e.g., 20, 80), poloxamers (e.g., 184, 188), pluronic polyols, and polyoxyethylene sorbitan monoethers (e.g., TWEEN®-20, TWEEN®-80). The nonionic surfactant may be present in a range of about 0.05 mg / mL to about 1.0 mg / mL, e.g., about 0.07 mg / mL to about 0.2 mg / mL.

[0138] Additional unclassified excipients include bulking agents (eg, starch), chelating agents (eg, EDTA), antioxidants (eg, ascorbic acid, methionine, vitamin E), and solvents.

[0139] Other pharmaceutical carriers Other pharmaceutically acceptable carriers that can be incorporated into the pharmaceutical compositions described herein include, but are not limited to, dextrose, sucrose, sorbitol, mannitol, starch, gum arabic, potassium phosphate, alginic acid, gelatin, potassium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methylcellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil. Compositions containing the antibodies described herein may further include lubricants, humectants, sweeteners, flavoring agents, emulsifiers, suspending agents, and preservatives. Details of suitable pharmaceutically acceptable carriers and formulations can be found in Remington's Pharmaceutical Sciences (19th ed., 1995); this reference is incorporated herein by reference.

[0140] [VII] Route and method of administration The anti-ITGA11 antibodies or antigen-binding fragments thereof described herein can be administered to mammalian subjects (e.g., humans) by a variety of routes, including orally, transdermally, subcutaneously, intranasally, intravenously, intramuscularly, intraocularly, intratumorally, parenterally, topically, intrathecally, and intracerebroventricularly, for the treatment of, for example, the diseases and conditions described herein (e.g., fibrosis and inflammatory disorders, or cancer). The most suitable route for administration in any given case will depend on the particular polypeptide being administered, the patient, the pharmaceutical formulation, the method of administration (e.g., timing and route of administration), the patient's age, weight, and sex, the severity of the disease being treated, the patient's diet, and the patient's excretion rate.

[0141] A physician of ordinary skill in the art can readily determine an effective amount of an anti-ITGA11 antibody or antigen-binding fragment thereof for administration to a mammalian subject (e.g., a human) in need thereof. For example, a physician can begin by prescribing a dose of the antibody described herein at a level lower than that required to achieve the desired therapeutic effect, and gradually increase the dose until the desired effect is achieved. Alternatively, a physician may begin a treatment regimen by administering a high dose of the antibody or antigen-binding fragment thereof described herein, and then administer progressively lower doses until a therapeutic effect is achieved. Generally, a suitable daily dose of an antibody or antigen-binding fragment thereof is that amount of the compound that is the minimum dose effective to produce a therapeutic effect. The antigen-binding fragments thereof described herein may be administered, for example, by injection (such as intravenous, intramuscular, intraperitoneal, or subcutaneous injection), optionally at a site near the target tissue site. The daily dosage of the therapeutic composition of the antibodies described herein may be administered as a single dose, or as two, three, four, five, six or more doses administered separately at appropriate intervals, optionally in unit dosage form, over the course of a day, week, month, or year, or as needed. While the antibodies described herein may be administered alone, they may also be administered as a pharmaceutical formulation in combination with excipients, carriers, and optionally, additional therapeutic agents.

[0142] An effective dose of an anti-ITGA11 antibody or antigen-binding fragment thereof described herein can range from about 0.0001 to about 100 mg / kg body weight per single (e.g., bolus), multiple, or continuous (e.g., continuous infusion) dose to achieve a serum concentration of 0.0001 to 5000 μg / mL, or any effective range or value depending on the condition being treated, the route of administration, and the age, weight, and condition of the subject. In certain embodiments, each dose can range from about 0.0001 mg to 500 mg / kg body weight. For example, the pharmaceutical compositions described herein can be administered at a daily dose ranging from 0.001 to 100 mg / kg body weight. The dose can be administered one or more times (e.g., 2 to 10 times) per day, week, month, or year to a mammalian subject (e.g., a human) in need thereof.

[0143] The anti-ITGA11 antibody or antigen-binding fragment thereof can be administered to a patient by continuous intravenous infusion or as a single bolus injection. The antibody or antigen-binding fragment thereof can be administered to a patient in an amount of, for example, 0.01 μg to about 5 g, and in a volume of, for example, 10 μL to 10 mL. The antibody or antigen-binding fragment thereof can be administered to a patient over a period of several minutes to several hours. For example, an antibody or antigen-binding fragment thereof described herein can be administered over a period of 5 minutes to 5 hours (e.g., 5 minutes, 10 minutes, 15 minutes, 20 minutes, 25 minutes, 30 minutes, 35 minutes, 40 minutes, 45 minutes, 50 minutes, 55 minutes, 60 minutes, 65 minutes, 70 minutes, 80 minutes, 90 minutes, 95 minutes, 100 minutes, 105 minutes, 110 minutes, 115 minutes, 120 minutes, 125 minutes, 130 minutes, 135 minutes, 140 minutes, 145 minutes, 150 minutes, 155 minutes, 160 minutes, 170 minutes, 180 minutes, 190 minutes, 210 minutes, 220 minutes, 230 minutes, 240 minutes, 250 minutes, 260 minutes, 270 minutes, 280 minutes, 290 minutes, 300 minutes, 310 minutes, 320 minutes, 330 minutes, 340 minutes, 350 minutes, 360 minutes, 370 minutes, 380 minutes, 390 minutes, 400 minutes, 410 minutes, 420 minutes, 430 minutes, 440 minutes, 450 minutes, 460 minutes, 470 minutes, 480 minutes, 490 minutes, 500 minutes, 51 , 165 minutes, 170 minutes, 175 minutes, 180 minutes, 185 minutes, 190 minutes, 195 minutes, 200 minutes, 205 minutes, 210 minutes, 215 minutes, 220 minutes, 225 minutes, 230 minutes, 235 minutes, 240 minutes, 245 minutes, 250 minutes, 255 minutes, 260 minutes, 265 minutes, 270 minutes, 275 minutes, 280 minutes, 285 minutes, 290 minutes, 295 minutes, or 300 minutes, or longer).

[0144] When an anti-ITGA11 antibody or antigen-binding fragment thereof is administered to a patient in combination with an additional therapeutic agent, the antibody or antigen-binding fragment thereof and the additional therapeutic agent may be co-administered to the patient by continuous intravenous infusion or bolus administration of the first agent, followed by continuous intravenous infusion or bolus administration of the second agent. The administration of the two agents may be simultaneous. Alternatively, administration of the antibody or antigen-binding fragment thereof may precede or follow administration of the additional therapeutic agent. In some embodiments, administration of the second agent (e.g., the antibody or antigen-binding fragment thereof) begins within about 5 minutes to about 4 weeks or longer of the end of administration of the first agent (e.g., the additional therapeutic agent). For example, administration of the second agent may begin within about 5 minutes, 10 minutes, 20 minutes, 30 minutes, 40 minutes, 50 minutes, 60 minutes, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, 13 hours, 14 hours, 15 hours, 16 hours, 17 hours, 18 hours, 19 hours, 20 hours, 21 hours, 22 hours, 23 hours, 24 hours, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 2 weeks, 3 weeks, 4 weeks or more of the end of administration of the first agent.

[0145] Therapeutic compositions can be administered by medical devices known in the art. For example, in one embodiment, the therapeutic compositions described herein can be administered by needleless hypodermic injection devices, such as those disclosed in U.S. Patent Nos. 5,399,163; 5,383,851; 5,312,335; 5,064,413; 4,941,880; 4,790,824; or 4,596,556. Examples of well-known implantable devices and modules useful with the compositions and methods described herein include the implantable microinfusion pump for rate-controlled administration disclosed in U.S. Patent No. 4,487,603; the therapeutic device for administering therapeutic drugs through the skin disclosed in U.S. Patent No. 4,486,194; the infusion pump for administering drugs at precise drip rates disclosed in U.S. Patent No. 4,447,233; the implantable variable flow infusion device for continuous drug delivery disclosed in U.S. Patent No. 4,447,224; the osmotic drug delivery system with multiple chamber compartments disclosed in U.S. Patent No. 4,439,196; and the osmotic drug delivery system disclosed in U.S. Patent No. 4,475,196. The contents of these patent documents are incorporated herein by reference. Many other similar implantable devices, delivery systems, and modules are known to those skilled in the art.

[0146] [VIII] A kit comprising an anti-ITGA11 antibody or an antigen-binding fragment thereof The present specification also includes kits containing anti-ITGA11 antibodies or antigen-binding fragments thereof. The kits provided herein may include any of the above-mentioned antibodies or antigen-binding fragments thereof, as well as polynucleotides encoding these polypeptides, vectors containing these polynucleotides, or cells (e.g., prokaryotic or eukaryotic cells) engineered to express and secrete the antibodies described herein.

[0147] The kits described herein may include reagents that can be used to produce the compositions described herein (e.g., anti-ITGA11 antibodies or antigen-binding fragments thereof). Optionally, the kits described herein may include a reagent capable of inducing expression of the antibody or antigen-binding fragment thereof in cells (e.g., mammalian cells), such as doxycycline or tetracycline. In other cases, the kits described herein may include a compound capable of binding and detecting a fusion protein comprising the antibody or antigen-binding fragment thereof and an epitope tag. For example, in such cases, the kits described herein may include maltose, glutathione, a nickel-containing complex, an anti-FLAG antibody, an anti-myc antibody, an anti-HA antibody, biotin, or streptavidin.

[0148] The kits described herein may also include reagents capable of directly detecting anti-ITGA11 antibodies or antigen-binding fragments thereof. Examples of such reagents include secondary antibodies that selectively recognize and bind to specific structural features within the Fc region of the antibodies or antigen-binding fragments described herein. The kits described herein may also include a secondary antibody that recognizes the Fc region of the antibody or antigen-binding fragment and is conjugated to a fluorescent molecule. These antibody / fluorophore conjugates provide a tool for analyzing the localization of the antibody or antigen-binding fragment, for example, in specific tissues or cultured mammalian cells, using established immunofluorescence techniques. In some embodiments, the kits described herein may include an additional fluorescent compound that exhibits a known subcellular localization pattern. These reagents can be used in combination with other antibody / fluorophore conjugates, for example, those that specifically recognize different receptors on the cell surface, to analyze the localization of the antibody or antigen-binding fragment relative to other cell surface proteins.

[0149] The kits described herein may also include reagents that can be used to analyze a patient's response to treatment with an antibody or antigen-binding fragment thereof described herein. For example, the kits described herein may include an antibody or antigen-binding fragment thereof and one or more reagents that can be used to measure the amount of regulatory T cells in a blood sample taken from a subject (e.g., a human) receiving treatment with an antibody described herein. The kits may include antibodies that selectively bind cell surface antigens presented by regulatory T cells, such as CD4 and CD25. Optionally, these antibodies may be labeled with a fluorescent dye, such as fluorescein or tetramethylrhodamine, to facilitate analysis of regulatory T cells by fluorescence-activated cell sorting (FACS) techniques known in the art. The kits described herein may optionally include one or more reagents that can be used to quantify tumor-reactive T lymphocytes to determine the effectiveness of the antibody or antigen-binding fragment thereof in restoring the proliferation of tumor-infiltrating lymphocytes. For example, the kits described herein may include antibodies that selectively bind cell surface markers on the surface of cytotoxic T cells, such as CD8 or CD3. Optionally, these antibodies may be labeled with a fluorescent molecule to allow quantification by FACS analysis.

[0150] The kits described herein may also include one or more reagents useful for determining the affinity and selectivity of the antibodies or antigen-binding fragments thereof described herein for one or more peptides derived from ITGA11. For example, the kit may include an anti-ITGA11 antibody or antigen-binding fragment thereof and a K of the antibodies described herein for one or more peptides that present an ITGA11 epitope in a conformation similar to that of the epitope in the native protein. DThe kit may include one or more reagents that can be used in an ELISA assay to determine ITGA11. The kit may include, for example, a microtiter plate with wells pre-conjugated to avidin, and may include a library of ITGA11-derived peptides, each of which is conjugated to a biotin moiety. Such a kit may optionally include a secondary antibody that specifically binds to the Fc region of an antibody or antigen-binding fragment thereof described herein, and the secondary antibody may be conjugated to an enzyme (e.g., horseradish peroxidase) that catalyzes a chemical reaction that produces light.

[0151] The kits described herein may also include an antibody or antigen-binding fragment thereof described herein, as well as reagents that can be conjugated to the antibody, etc., including those previously described (e.g., cytotoxic agents, fluorescent molecules, bioluminescent molecules, molecules comprising radioisotopes, molecules comprising chelating groups bound to paramagnetic ions, etc.). These kits may further include instructions on how to achieve conjugation of an antibody or antigen-binding fragment thereof described herein to a second molecule, such as those described above.

[0152] The kits described herein may also include a vector containing a polynucleotide encoding an antibody or antigen-binding fragment thereof, such as any of the vectors described herein. Alternatively, the kit may include mammalian cells (e.g., CHO cells) that have been genetically modified to express and secrete the antibody or antigen-binding fragment thereof, or a fragment thereof from the cell's nuclear genome. Such kits may also include instructions describing how expression of the antibody or antigen-binding fragment thereof from the polynucleotide can be induced, and may further include reagents (e.g., doxycycline or tetracycline) that can be used to promote transcription of these polynucleotides. Such kits may be useful for producing the antibodies or antigen-binding fragments thereof described herein.

[0153] Other kits described herein may include tools for engineering prokaryotic or eukaryotic cells (e.g., CHO cells or BL21(DE3) E. coli cells) to express and secrete an antibody or antigen-binding fragment thereof described herein from the cell's nuclear genome. For example, the kit may include CHO cells maintained in an appropriate medium and optionally frozen according to methods known in the art. The kit may also provide a vector containing a polynucleotide encoding a nuclease (e.g., a CRISPER / Cas, zinc finger nuclease, TALEN, ARCUS® nuclease, etc. described herein) as well as reagents for expressing the nuclease in the cell. The kit can further provide tools for modifying the polynucleotide encoding the nuclease to allow for alteration of the nuclease's DNA sequence to direct cleavage of a specific target DNA sequence of interest. Examples of such tools include primers for amplification and site-specific mutagenesis of a polynucleotide encoding a nuclease of interest. The kit may also include a restriction enzyme that can be used to selectively excise the nuclease-encoding polynucleotide from the vector and then reintroduce the modified polynucleotide into the vector once the user has modified the gene. Such a kit may also include a DNA ligase that can be used to catalyze the formation of a covalent phosphodiester bond between the modified nuclease-encoding polynucleotide and the target vector. The kits described herein may also provide a polynucleotide encoding an antibody or antigen-binding fragment thereof, as well as instructions that describe methods that can be used to selectively cleave a specific DNA sequence in the genome of a cell to integrate the polynucleotide encoding the antibody or antigen-binding fragment thereof into the genome at this site. Optionally, the kit may provide a polynucleotide encoding a fusion protein comprising the antibody or antigen-binding fragment thereof and an additional polypeptide, such as those described herein.

[0154] Example The following examples are presented to provide one of ordinary skill in the art with an illustration of how the compositions and methods described herein can be used, made, and evaluated, and are intended to be purely exemplary of the disclosure and are not intended to limit the scope of the invention disclosed by the inventors.

[0155] Example 1: Generation and purification of anti-ITGA11 antibodies Anti-ITGA11 antibody was administered at 10 10 The naive human scFv library generated by IONTAS with a unique clonal diversity of 1000 kJ / s was used. Two rounds of phage display selection were performed on this library with a total of eight selection elements, including selections on various combinations of recombinant soluble human ITGA11B1, recombinant mouse ITGA11B1, cell-expressed human ITGA11B1, and cell-expressed mouse ITGA11B1. Negative selection was also included with cell-expressed human ITGA2B1 and recombinant ITGA2B1. Each selection element was performed in the presence or absence of 1 mM MnCl2 for all eight selection elements, including outputs with kappa light chains, and all eight selection elements, including outputs with lambda light chains. After the final round of selection and negative selection for cell-expressed ITGA11, all selection outputs showed enrichment after the second round of selection.

[0156] After isolation of the scFv phage display selection outputs, they were converted into a mammalian display human IgG1 format. These converted outputs were grouped into four mammalian display libraries, and library sizes were determined using methods known in the art. These libraries were prepared for mammalian display transformation and selection in HEK293 cells. 100 million HEK293 cells were transformed with 20 μg of the library DNA mixture. The day after transformation, cells from each transformant were incubated with an anti-hu Fc-PE-labeled antibody to assess the transient expression level of cell surface IgG1. In the first round of pre-mammalian cell display (MCD), the transformed libraries were subjected to MACS (Miltenyi Midi MACS separator) sorting to enrich for cells expressing high levels of human IgG1 (detected by anti-Fc-PE-conjugated anti-PE magnetic beads bound to cells expressing human IgG1). Results for each of the mammalian display libraries showed enrichment for those that bind to human ITGA11B1. In the first round of MCD, all MCD libraries were incubated with 0.1 nM biotinylated human ITGA11B1 and sorted using FACS. Unlabeled human ITGA10B1 was included to reduce the number of selected clones that bound it. 30 million cells were sorted per library. The top 1.5–4.9% of human ITGA11B1 binders at 0.1 nM were collected and added to cell culture for the second round of sorting. This process was repeated in the second round of sorting, increasing the stringency of human ITGA11 binding to enrich for higher affinity binders and also including biotinylated ITGA10 as a specificity selection.

[0157] The mammalian display output cell population from the second round of sorting was subcloned into a soluble human IgG4 (S228P) expression vector. Plasmid DNA preparations for the output population expression vector were prepared and propagated in E. coli using methods known in the art. Colonies were picked from all outputs and distributed into ten 96-deep-well plates in 1.2 ml of LB medium and kanamycin and incubated overnight at 37°C with shaking. Each plate contained 90 antibody clones, one positive control, one negative control antibody, and two blank wells. Plasmid DNA for each well of all ten plates was extracted and purified using the MagBind kit. The purified plasmid DNA was used to transform ExpiHEK293 cells for expression of each soluble antibody clone. Conditioned medium containing each antibody was used for the first screening experiment. All human IgG4 antibody clones, including supernatants from 10 plates, were evaluated by ELISA for binding to CHO cells expressing human ITGA11B1 and to recombinant human ITGA11B1 protein. Of the 900 clones screened, 61 unique clones bound to CHO human ITGA11B1 cells while showing minimal binding to CHO human ITGA10B1 or human ITGA2B1. These 61 antibody clones were transiently expressed in Expi293 cells in 24-deep-well plates, and the antibodies were purified using protein A affinity chromatography according to standard methods known to those skilled in the art.

[0158] The 61 purified antibody hits were then functionally evaluated using a hierarchical flow scheme to measure their binding ability to cells expressing human ITGA11B1 at 34 nM and their inhibitory ability to inhibit human ITGA11B1 binding to collagen at 68 nM. Of the 61 hits, 20 antibodies were identified that inhibited type I collagen binding to human ITGA11B1 by at least 50% at 68 nM. All of these antibodies showed measurable binding to human ITGA11B and cynomolgus monkey ITGA11B1, and 17 of the 20 antibodies bound to mouse ITGA11B1.

[0159] Example 2: Characterization of the binding of anti-ITGA11 antibodies Antibody binding to C2C12 cells expressing human ITGA11 or CHO cells expressing mouse ITGA11 was performed by incubating the cells with the antibody at a concentration of 34 nM in binding medium (PBS, 10% normal goat serum, 2% rabbit serum, and 1% BSA) for 1 hour at 4°C. The cells were washed in binding medium, and binding was detected using goat anti-human Fc BV421 (Jackson ImmunoLabs). After washing, the cells were resuspended in binding medium, and binding was assessed by flow cytometry. 226 antibodies were screened, and the binding characteristics of 10 antibodies are shown below. The results are shown in Table 2 and are expressed as mean fluorescence intensity (MFI).

[0160] [Table 2]

[0161] Example 3: Anti-ITGA11 antibody inhibits collagen binding to cell-expressed ITGA11 Collagen conjugated to fluorescently labeled beads was incubated with anti-ITGA11 antibody and mouse C2C12 cells expressing human ITGA11 for 1 hour in a binding medium containing DMEM, 10% normal goat serum, and 2% normal rabbit serum. The cells were trypsinized, and unbound collagen (rat tail type I collagen, Corning Labs)-coated beads (fluorescent YG carboxylated microspheres, 2 microns, Polysciences) were washed off the cells twice with PBS. The cells were resuspended in ice-cold PBS containing 2% fetal bovine serum and 1 mM EDTA, and the number of bound beads was counted by flow cytometry. The results, shown in Figures 1, 2, and 3, are expressed as the percentage of bound beads compared to the total number of beads incubated with the cells. MOPC21 mAb IgG1 was used as a negative control.

[0162] The anti-ITGA11 antibodies FIB-918-1, FIB-918-2, FIB-918-3, and FIB-918-4 (Figure 1); FIB-918-5, FIB-918-6, FIB-918-7, and FIB-918-8 (Figure 2); and FIB-918-9 and FIB-918-10 (Figure 3) inhibited collagen binding to cell-expressed ITGA11 to different extents based on the range of IC50s observed.

[0163] Other embodiments While the present disclosure has been described in connection with specific embodiments thereof, it will be understood that further modifications are possible, and that the present application is intended to cover all variations, adaptations, or applications which may conform to the essential features described above and which fall within the scope of the claims, which generally follow the principles of the present disclosure and may include such variations, adaptations, or applications as may be evolved from the present disclosure within the scope of known or customary practice in the art to which the present disclosure pertains. Other embodiments are within the scope of the claims.

Claims

1. An antibody or antigen-binding fragment thereof that specifically binds to integrin α11 (ITGA11), a complementarity determining region (CDR) heavy chain 1 (CDR-H1) comprising the amino acid sequence of GFTFSSYA (SEQ ID NO: 1), GFTFSNAW (SEQ ID NO: 9), GFTFSSYS (SEQ ID NO: 14), GYTFTDYY (SEQ ID NO: 28), GFTFSDYW (SEQ ID NO: 36), or GFMFDTHA (SEQ ID NO: 46); a complementarity-determining region (CDR) heavy chain 2 (CDR-H2) comprising the amino acid sequence of ISGSGGST (SEQ ID NO: 2), ISSSSSSTI (SEQ ID NO: 15), FDPEDGET (SEQ ID NO: 29), or ISGSGGSI (SEQ ID NO: 74); Complementarity determining region (CDR) heavy chain 3 (CDR-H3) comprising the amino acid sequences of AKDLDWSGHDAFDI (SEQ ID NO: 3), ARDRGYSYSETSNDAFDI (SEQ ID NO: 10), ARGPDLSDYFDY (SEQ ID NO: 16), AKDPRGSGRDDAFDI (SEQ ID NO: 20), AKDPTTMTTDAFDI (SEQ ID NO: 25), ATLDYRGVVYFDY (SEQ ID NO: 30), AKDLLWAARDAFDI (SEQ ID NO: 37), AKQTVTSADDYFDY (SEQ ID NO: 43), ARSGETAGTDYFDY (SEQ ID NO: 48); Complementarity-determining region (CDR) light chain 1 (CDR-L1) comprising the amino acid sequences of QSISSY (SEQ ID NO: 4), QTIGSY (SEQ ID NO: 21), SGSIASNY (SEQ ID NO: 31), QGINDF (SEQ ID NO: 40), and QSVSSSY (SEQ ID NO: 49); A complementarity-determining region (CDR) light chain 2 (CDR-L2) comprising the amino acid sequence of AAS (SEQ ID NO: 5), GAS (SEQ ID NO: 22), or EDK (SEQ ID NO: 32); and a complementarity-determining region (CDR) light chain 3 (CDR-L3) comprising the amino acid sequence of QQTYSTPLT (SEQ ID NO: 6), QQSYSTPFT (SEQ ID NO: 11), QQSYSTPLT (SEQ ID NO: 17), QSYDSSNHWV (SEQ ID NO: 33), or QQDYNSPYT (SEQ ID NO: 50); An antibody or antigen-binding fragment thereof comprising:

2. The antibody or antigen-binding fragment thereof of claim 1, wherein CDR-H1 comprises the amino acid sequence of GFTFSSYA (SEQ ID NO: 1).

3. The antibody or antigen-binding fragment thereof of claim 1 or 2, wherein CDR-H2 comprises the amino acid sequence ISGSGGST (SEQ ID NO: 2).

4. The antibody or antigen-binding fragment thereof according to any one of claims 1 to 3, wherein CDR-L1 comprises the amino acid sequence of QSISSY (SEQ ID NO: 4).

5. The antibody or antigen-binding fragment thereof according to any one of claims 1 to 4, wherein CDR-L2 comprises the amino acid sequence of AAS (SEQ ID NO: 5).

6. The antibody or antigen-binding fragment thereof according to any one of claims 1 to 5, wherein CDR-L3 comprises the amino acid sequence QQSYSTPFT (SEQ ID NO: 11).

7. CDR-H1 comprising the amino acid sequence of GFTFSSYA (SEQ ID NO: 1); CDR-H2 comprising the amino acid sequence ISGSGGST (SEQ ID NO: 2); CDR-H3 comprising the amino acid sequence of AKDLDWSGHDAFDI (SEQ ID NO: 3); CDR-L1 comprising the amino acid sequence of QSISSY (SEQ ID NO: 4); CDR-L2 comprising the amino acid sequence of AAS (SEQ ID NO: 5); and QQTYSTPLT (SEQ ID NO: 6) CDR-L3, The antibody or antigen-binding fragment thereof of claim 1, comprising:

8. a heavy chain variable domain comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 7; and a light chain variable domain comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 8; The antibody or antigen-binding fragment thereof of claim 7, comprising:

9. A heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 7; and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 8; The antibody or antigen-binding fragment thereof of claim 8, comprising:

10. CDR-H1 comprising the amino acid sequence of GFTFSNAW (SEQ ID NO: 9); CDR-H2 comprising the amino acid sequence ISGSGGST (SEQ ID NO: 2); CDR-H3 comprising the amino acid sequence of ARDRGYSYSETSNDAFDI (SEQ ID NO: 10); CDR-L1 comprising the amino acid sequence of QSISSY (SEQ ID NO: 4); CDR-L2 comprising the amino acid sequence of AAS (SEQ ID NO: 5); and CDR-L3 comprising the amino acid sequence of QQSYSTPFT (SEQ ID NO: 11); The antibody or antigen-binding fragment thereof of claim 1, comprising:

11. a heavy chain variable domain comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 12; and a light chain variable domain comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 13; The antibody or antigen-binding fragment thereof of claim 10, comprising:

12. A heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 12; and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 13; The antibody or antigen-binding fragment thereof of claim 11, comprising:

13. CDR-H1 comprising the amino acid sequence of GFTFSSYS (SEQ ID NO: 14); CDR-H2 comprising the amino acid sequence of ISSSSSSTI (SEQ ID NO: 15); CDR-H3 comprising the amino acid sequence of ARGPDLSDYFDY (SEQ ID NO: 16); CDR-L1 comprising the amino acid sequence of QSISSY (SEQ ID NO: 4); CDR-L2 comprising the amino acid sequence of AAS (SEQ ID NO: 5); and CDR-L3 comprising the amino acid sequence of QQSYSTPLT (SEQ ID NO: 17); The antibody or antigen-binding fragment thereof of claim 1, comprising:

14. a heavy chain variable domain comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 12; and a light chain variable domain comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 13; The antibody or antigen-binding fragment thereof of claim 10, comprising:

15. A heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 12; and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 13; The antibody or antigen-binding fragment thereof of claim 11, comprising:

16. CDR-H1 comprising the amino acid sequence of GFTFSSYA (SEQ ID NO: 1); CDR-H2 comprising the amino acid sequence ISGSGGST (SEQ ID NO: 2); CDR-H3 comprising the amino acid sequence of AKDPRGSGRDDAFDI (SEQ ID NO: 20); CDR-L1 comprising the amino acid sequence of QTIGSY (SEQ ID NO: 21); CDR-L2 comprising the amino acid sequence of GAS (SEQ ID NO: 22); and CDR-L3 comprising the amino acid sequence of QQSYSTPFT (SEQ ID NO: 11); The antibody or antigen-binding fragment thereof of claim 1, comprising:

17. a heavy chain variable domain comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:23; and a light chain variable domain comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 24; 17. The antibody or antigen-binding fragment thereof of claim 16, comprising:

18. A heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 23; and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 24; The antibody or antigen-binding fragment thereof of claim 15, comprising:

19. CDR-H1 comprising the amino acid sequence of GFTFSSYA (SEQ ID NO: 1); CDR-H2 comprising the amino acid sequence ISGSGGST (SEQ ID NO: 2); CDR-H3 comprising the amino acid sequence of AKDPTTMTTDAFDI (SEQ ID NO: 25); CDR-L1 comprising the amino acid sequence of QSISSY (SEQ ID NO: 4); CDR-L2 comprising the amino acid sequence of AAS (SEQ ID NO: 5); and CDR-L3 comprising the amino acid sequence of QQSYSTPFT (SEQ ID NO: 11); The antibody or antigen-binding fragment thereof of claim 1, comprising:

20. a heavy chain variable domain comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:26; and a light chain variable domain comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 27; 20. The antibody or antigen-binding fragment thereof of claim 19, comprising:

21. A heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 26; and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 27; 21. The antibody or antigen-binding fragment thereof of claim 20, comprising:

22. CDR-H1 comprising the amino acid sequence of GYTFTDYY (SEQ ID NO: 28); CDR-H2 comprising the amino acid sequence of FDPEDGET (SEQ ID NO: 29); CDR-H3 comprising the amino acid sequence of ATLDYRGVVYFDY (SEQ ID NO: 30); CDR-L1 comprising the amino acid sequence of SGSIASNY (SEQ ID NO: 31); CDR-L2 comprising the amino acid sequence of EDK (SEQ ID NO: 32); and CDR-L3 comprising the amino acid sequence of QSYDSSNHWV (SEQ ID NO: 33); The antibody or antigen-binding fragment thereof of claim 1, comprising:

23. a heavy chain variable domain comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 34; and a light chain variable domain comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 35; 23. The antibody or antigen-binding fragment thereof of claim 22, comprising:

24. A heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 34; and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 35; 24. The antibody or antigen-binding fragment thereof of claim 23, comprising:

25. CDR-H1 comprising the amino acid sequence of GFTFSDYW (SEQ ID NO: 36); CDR-H2 comprising the amino acid sequence ISGSGGST (SEQ ID NO: 2); CDR-H3 comprising the amino acid sequence of AKDLLWAARDAFDI (SEQ ID NO: 37); CDR-L1 comprising the amino acid sequence of QSISSY (SEQ ID NO: 4); CDR-L2 comprising the amino acid sequence of AAS (SEQ ID NO: 5); and CDR-L3 comprising the amino acid sequence of QQSYSTPFT (SEQ ID NO: 11); The antibody or antigen-binding fragment thereof of claim 1, comprising:

26. a heavy chain variable domain comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 38; and a light chain variable domain comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 39; 26. The antibody or antigen-binding fragment thereof of claim 25, comprising:

27. A heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 38; and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 39; 27. The antibody or antigen-binding fragment thereof of claim 26, comprising:

28. CDR-H1 comprising the amino acid sequence of GFTFSSYA (SEQ ID NO: 1); CDR-H2 comprising the amino acid sequence ISGSGGST (SEQ ID NO: 2); CDR-H3 comprising the amino acid sequence of AKDLLWAARDAFDI (SEQ ID NO: 37); CDR-L1 comprising the amino acid sequence of QGINDF (SEQ ID NO: 40); CDR-L2 comprising the amino acid sequence of AAS (SEQ ID NO: 5); and CDR-L3 comprising the amino acid sequence of QQSYSTPLT (SEQ ID NO: 17); The antibody or antigen-binding fragment thereof of claim 1, comprising:

29. a heavy chain variable domain comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:41; and a light chain variable domain comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 42; 29. The antibody or antigen-binding fragment thereof of claim 28, comprising:

30. A heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 41; and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 42; 30. The antibody or antigen-binding fragment thereof of claim 29, comprising:

31. CDR-H1 comprising the amino acid sequence of GFTFSNAW (SEQ ID NO: 9); CDR-H2 comprising the amino acid sequence ISGSGGST (SEQ ID NO: 2); CDR-H3 comprising the amino acid sequence of AKQTVTSADDYFDY (SEQ ID NO: 43); CDR-L1 comprising the amino acid sequence of QSISSY (SEQ ID NO: 4); CDR-L2 comprising the amino acid sequence of AAS (SEQ ID NO: 5); and CDR-L3 comprising the amino acid sequence of QQSYSTPFT (SEQ ID NO: 11); The antibody or antigen-binding fragment thereof of claim 1, comprising:

32. a heavy chain variable domain comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:44; and a light chain variable domain comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 45; 32. The antibody or antigen-binding fragment thereof of claim 31 , comprising:

33. A heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 44; and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 45; 33. The antibody or antigen-binding fragment thereof of claim 32, comprising:

34. CDR-H1 comprising the amino acid sequence of GFMFDTHA (SEQ ID NO: 46); CDR-H2 comprising the amino acid sequence of ISGSGGSI (SEQ ID NO: 47); CDR-H3 comprising the amino acid sequence of ARSGETAGTDYFDY (SEQ ID NO: 48); CDR-L1 comprising the amino acid sequence of QSVSSSY (SEQ ID NO: 49); CDR-L2 comprising the amino acid sequence of GAS (SEQ ID NO: 22); and CDR-L3 comprising the amino acid sequence of QQDYNSPYT (SEQ ID NO: 50); The antibody or antigen-binding fragment thereof of claim 1, comprising:

35. a heavy chain variable domain comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 51; and a light chain variable domain comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 52; 35. The antibody or antigen-binding fragment thereof of claim 34, comprising:

36. A heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 51; and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 52; 36. The antibody or antigen-binding fragment thereof of claim 35, comprising:

37. The antibody or antigen-binding fragment thereof of claim 1, comprising a heavy chain variable domain comprising an amino acid sequence having at least 90% sequence identity to any one of the amino acid sequences of SEQ ID NO: 7, 12, 18, 23, 26, 34, 38, 41, 44, or 51.

38. The antibody or antigen-binding fragment thereof of claim 1 or 36, comprising a light chain variable domain comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs: 8, 13, 19, 24, 35, 39, 42, 45, or 52.

39. Monoclonal antibodies or antigen-binding fragments thereof, polyclonal antibodies or antigen-binding fragments thereof, human antibodies or antigen-binding fragments thereof, humanized antibodies or antigen-binding fragments thereof, primatized antibodies or antigen-binding fragments thereof, bispecific antibodies or antigen-binding fragments thereof, multispecific antibodies or antigen-binding fragments thereof, dual variable immunoglobulin domains, monovalent antibodies or antigen-binding fragments thereof, chimeric antibodies or antigen-binding fragments thereof, single-chain Fv molecules (scFv), diabodies, triabodies, nanobodies, antibody-like protein scaffolds, domain antibodies, Fv fragments, Fab fragments, F(ab') ２ 39. The antibody or antigen-binding fragment thereof of any one of claims 1 to 38, wherein the antibody or antigen-binding fragment thereof is selected from the group consisting of a fusion protein, a fusion protein molecule, and a tandem scFv (taFv).

40. 40. The antibody or antigen-binding fragment thereof of claim 39, which is a human antibody, a humanized antibody, or a chimeric antibody or antigen-binding fragment thereof.

41. The antibody or antigen-binding fragment thereof according to any one of claims 1 to 38, which specifically binds to a heterodimer of ITGA11 and ITGB1 (ITGA11B1 heterodimer).

42. A polynucleotide encoding the antibody or antigen-binding fragment thereof according to any one of claims 1 to 41.

43. A vector comprising the polynucleotide of claim 42.

44. 44. The vector of claim 43 which is an expression vector.

45. 44. The vector of claim 43, wherein the expression vector is a eukaryotic expression vector.

46. 46. ​​The vector of claim 45 which is a viral vector.

47. 47. The vector of claim 46, wherein the viral vector is selected from the group consisting of adenovirus (Ad), retrovirus, poxvirus, adeno-associated virus, baculovirus, herpes simplex virus, and vaccinia virus.

48. A host cell comprising the vector according to any one of claims 43 to 47.

49. 49. A pharmaceutical composition comprising the antibody or antigen-binding fragment thereof of any one of claims 1 to 41, the polynucleotide of claim 42, the vector of any one of claims 43 to 47, or the host cell of claim 48, and a pharmaceutically acceptable carrier or excipient.

50. 50. A kit comprising an agent selected from the antibody or antigen-binding fragment thereof of any one of claims 1 to 41, the polynucleotide of claim 42, the vector of any one of claims 43 to 47, the host cell of claim 48, or the pharmaceutical composition of claim 49.

51. 50. A method of treating a subject having or at risk of developing a disorder, comprising administering to the subject the antibody or antigen-binding fragment thereof of any one of claims 1 to 41, the polynucleotide of claim 42, the vector of any one of claims 43 to 47, the host cell of claim 48, or the pharmaceutical composition of claim 49.

52. 52. The method of claim 51, wherein the disorder is fibrosis.

53. 53. The method of claim 52, wherein the fibrosis is selected from liver fibrosis, kidney fibrosis, skin fibrosis, cardiac fibrosis, vascular fibrosis, ocular fibrosis, bone marrow fibrosis, pulmonary fibrosis, glomerulonephritis, heart failure, scleroderma, excessive scar tissue after surgery or device insertion, trauma or burns, progressive kidney disease, valvular heart disease, hypertensive heart disease, articular and periarticular fibrosis, bone marrow fibrosis, ocular / vitreous fibrosis, intestinal fibrosis and stenosis, peritoneal and retroperitoneal fibrosis, pancreatic fibrosis, nephrogenic systemic fibrosis, and primary sclerosing cholangitis.

54. 52. The method of claim 51, wherein the disorder is an inflammatory disorder.

55. Inflammatory disorders include asthma, airway inflammation, airway hyperreactivity, and airway hyperresponsiveness.

55. The method of claim 54, wherein the inflammatory disease is selected from hyperresponsiveness, rhinosinusitis, rhinosinusitis with polyps, nasal polyps, arthritis, eosinophilic inflammation, mast cell-mediated inflammatory diseases, sepsis, septic shock, seronegative enthesitis / arthritis (SEA) syndrome, osteoporosis, eosinophilic esophagitis, scleroderma, dermatitis, atopic dermatitis, allergic rhinitis, bullous pemphigoid, urticaria, inflammation of the cartilage, polymyalgia rheumatica, Wegener's granulomatosis, Behcet's disease, myositis, polymyositis, dermatomyositis, vasculitis, arteritis, diabetic nephropathy, interstitial cystitis, graft versus host disease (GVHD), inflammatory conditions of the gastrointestinal tract, and inflammatory pulmonary conditions.

56. 52. The method of claim 51, wherein the disorder is cancer.

57. 57. The method of claim 56, wherein the cancer is selected from breast cancer, colorectal cancer, liver cancer, kidney cancer, liver cancer, lung cancer, pancreatic cancer, cancer of the gastrointestinal tract, melanoma, ovarian cancer, prostate cancer, cervical cancer, bladder cancer, glioblastoma, head and neck cancer, and bile duct cancer.

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