Antibodies binding to integrin alpha11 and uses thereof

Abstract

The present disclosure provides an antibody, or an antigen-binding fragment thereof, that specifically binds to integrin alpha 11 (ITGA11). The present disclosure also provides methods for treating a condition (e.g., a fibrotic condition, an inflammatory condition, or cancer) in a subject. The disclosure includes related pharmaceutical compositions, polynucleotides, vectors, host cells, methods of preparation, methods of treatment, methods of diagnosis, and kits.

Description

[0001] Related Applications

[0002] This Patent Cooperation Treaty application claims the benefit of the priority of U.S. Provisional Application No. 63 / 380,668, filed Oct. 24, 2022, which is hereby incorporated by reference in its entirety.

[0003] Background

[0004] The interaction between cells and their extracellular microenvironment is mainly mediated by a family of cell surface receptors called integrins, which are considered key molecules involved in myofibroblast differentiation. Integrins are heterodimeric transmembrane receptors containing α and β subunits, and the α and β subunits can combine to form 24 different integrin heterodimers. Integrins regulate cytoskeletal dynamics, thereby affecting many crucial 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)). Integrin α-chain α11 (ITGA11) interacts with integrin β-chain β1 to form the α11β1 heterodimer. α11β1 preferentially binds to type I collagen and has been shown to be expressed on cultured embryonic fibroblasts, cardiac fibroblasts, and on activated hepatic, pulmonary, and renal myofibroblasts. α11β1 has been shown to be upregulated by TGFβ, and α11β1 regulates the differentiation of embryonic mesenchymal cells on a collagen matrix. α11β1 is also associated with inducing tumor growth and the metastatic potential of small cell lung cancer cells. ITGA11 is overexpressed in fibrotic disease tissues, including the lungs of IPF patients and the kidneys of patients with chronic allograft nephropathy. Thus, ITGA11 is a target for therapeutic intervention, and therapeutic modalities that bind to ITGA11 are needed.

[0005] Overview

[0006] The present disclosure provides antibodies or antigen-binding fragments thereof that specifically bind to integrin α11 (ITGA11). The present disclosure also provides methods for treating ITGA11-related disorders (such as fibrotic disorders, inflammatory disorders, or cancers) in a subject. The present disclosure also includes related pharmaceutical compositions, polynucleotides, vectors, host cells, preparation methods, treatment methods, diagnostic methods, and kits.

[0007] In one aspect, the present disclosure provides an antibody or an antigen-binding fragment thereof that specifically binds to integrin α11 (ITGA11), wherein the antibody or the antigen-binding fragment thereof comprises:

[0008] Complementary determining region (CDR)-heavy chain 1 (CDR-H1), which comprises the amino acid sequence 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);

[0009] Complementary determining region (CDR)-heavy chain 2 (CDR-H2), which comprises the amino acid sequence ISGSGGST (SEQ ID NO:2), ISSSSSTI (SEQ ID NO:15), FDPEDGET (SEQ ID NO:29) or ISGSGGSI (SEQ ID NO:74);

[0010] Complementary determining region (CDR)-heavy chain 3 (CDR-H3), which comprises the amino acid sequence 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);

[0011] Complementary determining region (CDR)-light chain 1 (CDR-L1), which comprises the amino acid sequence QSISSY (SEQ ID NO:4), QTIGSY (SEQ ID NO:21), SGSIASNY (SEQ ID NO:31), QGINDF (SEQ ID NO:40), QSVSSSY (SEQ ID NO:49);

[0012] Complementary determining region (CDR)-light chain 2 (CDR-L2), which comprises the amino acid sequence AAS (SEQ ID NO:5), GAS (SEQ ID NO:22) or EDK (SEQ ID NO:32); and

[0013] Complementary determining region (CDR)-light chain 3 (CDR-L3), which comprises the amino acid sequence 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).

[0014] In some embodiments, the antibody or antigen-binding fragment comprises CDR-H1 containing the amino acid sequence GFTFSSYA (SEQ ID NO:1).

[0015] In some embodiments, the antibody or antigen-binding fragment comprises CDR-H2 containing the amino acid sequence ISGSGGST (SEQ ID NO:2).

[0016] In some embodiments, the antibody or antigen-binding fragment comprises CDR-L1 containing the amino acid sequence QSISSY (SEQ ID NO:4).

[0017] In some embodiments, the antibody or antigen-binding fragment comprises CDR-L2 containing the amino acid sequence AAS (SEQ ID NO:5).

[0018] In some embodiments, the antibody or antigen-binding fragment comprises CDR-L3 containing the amino acid sequence QQSYSTPFT (SEQ ID NO:11).

[0019] In some embodiments, the antibody or antigen-binding fragment thereof comprises: CDR-H1 comprising the amino acid sequence GFTFSSYA (SEQ ID NO:1); CDR-H2 comprising the amino acid sequence ISGSGGST (SEQ ID NO:2); CDR-H3 comprising the amino acid sequence AKDLDWSGHDAFDI (SEQ ID NO:3); CDR-L1 comprising the amino acid sequence QSISSY (SEQ ID NO:4); CDR-L2 comprising the amino acid sequence AAS (SEQ ID NO:5); and CDR-L3 comprising the amino acid sequence 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.

[0020] In some embodiments, the antibody or antigen-binding fragment thereof comprises: CDR-H1 comprising the amino acid sequence GFTFSNAW (SEQ ID NO:9); CDR-H2 comprising the amino acid sequence ISGSGGST (SEQ ID NO:2); CDR-H3 comprising the amino acid sequence ARDRGYSYSETSNDAFDI (SEQ ID NO:10); CDR-L1 comprising the amino acid sequence QSISSY (SEQ ID NO:4); CDR-L2 comprising the amino acid sequence AAS (SEQ ID NO:5); and CDR-L3 comprising the amino acid sequence 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.

[0021] In some embodiments, the antibody or antigen-binding fragment thereof comprises: CDR-H1 comprising the amino acid sequence GFTFSSYS (SEQ ID NO:14); CDR-H2 comprising the amino acid sequence ISSSSSTI (SEQ ID NO:15); CDR-H3 comprising the amino acid sequence ARGPDLSDYFDY (SEQ ID NO:16); CDR-L1 comprising the amino acid sequence QSISSY (SEQ ID NO:4); CDR-L2 comprising the amino acid sequence AAS (SEQ ID NO:5); and CDR-L3 comprising the amino acid sequence 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.

[0022] In some embodiments, the antibody or antigen-binding fragment thereof comprises: CDR-H1 comprising the amino acid sequence GFTFSSYA (SEQ ID NO:1); CDR-H2 comprising the amino acid sequence ISGSGGST (SEQ ID NO:2); CDR-H3 comprising the amino acid sequence AKDPRGSGRDDAFDI (SEQ ID NO:20); CDR-L1 comprising the amino acid sequence QTIGSY (SEQ ID NO:21); CDR-L2 comprising the amino acid sequence GAS (SEQ ID NO:22); and CDR-L3 comprising the amino acid sequence 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.

[0023] In some embodiments, the antibody or antigen-binding fragment thereof comprises: CDR-H1 comprising the amino acid sequence GFTFSSYA (SEQ ID NO:1); CDR-H2 comprising the amino acid sequence ISGSGGST (SEQ ID NO:2); CDR-H3 comprising the amino acid sequence AKDPTTMTTDAFDI (SEQ ID NO:25); CDR-L1 comprising the amino acid sequence QSISSY (SEQ ID NO:4); CDR-L2 comprising the amino acid sequence AAS (SEQ ID NO:5); and CDR-L3 comprising the amino acid sequence 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.

[0024] In some embodiments, the antibody or antigen-binding fragment thereof comprises: CDR-H1 comprising the amino acid sequence GYTFTDYY (SEQ ID NO:28); CDR-H2 comprising the amino acid sequence FDPEDGET (SEQ ID NO:29); CDR-H3 comprising the amino acid sequence ATLDYRGVVYFDY (SEQ ID NO:30); CDR-L1 comprising the amino acid sequence SGSIASNY (SEQ ID NO:31); CDR-L2 comprising the amino acid sequence EDK (SEQ ID NO:32); and CDR-L3 comprising the amino acid sequence 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.

[0025] In some embodiments, the antibody or antigen-binding fragment thereof comprises: CDR-H1 comprising the amino acid sequence GFTFSDYW (SEQ ID NO:36); CDR-H2 comprising the amino acid sequence ISGSGGST (SEQ ID NO:2); CDR-H3 comprising the amino acid sequence AKDLLWAARDAFDI (SEQ ID NO:37); CDR-L1 comprising the amino acid sequence QSISSY (SEQ ID NO:4); CDR-L2 comprising the amino acid sequence AAS (SEQ ID NO:5); and CDR-L3 comprising the amino acid sequence 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.

[0026] In some embodiments, the antibody or antigen-binding fragment thereof comprises: CDR-H1 comprising the amino acid sequence GFTFSSYA (SEQ ID NO:1); CDR-H2 comprising the amino acid sequence ISGSGGST (SEQ ID NO:2); CDR-H3 comprising the amino acid sequence AKDLLWAARDAFDI (SEQ ID NO:37); CDR-L1 comprising the amino acid sequence QGINDF (SEQ ID NO:40); CDR-L2 comprising the amino acid sequence AAS (SEQ ID NO:5); and CDR-L3 comprising the amino acid sequence 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.

[0027] In some embodiments, the antibody or antigen-binding fragment thereof comprises: CDR-H1 comprising the amino acid sequence GFTFSNAW (SEQ ID NO:9); CDR-H2 comprising the amino acid sequence ISGSGGST (SEQ ID NO:2); CDR-H3 comprising the amino acid sequence AKQTVTSADDYFDY (SEQ ID NO:43); CDR-L1 comprising the amino acid sequence QSISSY (SEQ ID NO:4); CDR-L2 comprising the amino acid sequence AAS (SEQ ID NO:5); and CDR-L3 comprising the amino acid sequence 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.

[0028] In some embodiments, the antibody or antigen-binding fragment thereof comprises: CDR-H1 comprising the amino acid sequence GFMFDTHA (SEQ ID NO: 46); CDR-H2 comprising the amino acid sequence ISGSGGSI (SEQ ID NO: 47); CDR-H3 comprising the amino acid sequence ARSGETAGTDYFDY (SEQ ID NO: 48); CDR-L1 comprising the amino acid sequence QSVSSSY (SEQ ID NO: 49); CDR-L2 comprising the amino acid sequence GAS (SEQ ID NO: 22); and CDR-L3 comprising the amino acid sequence 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.

[0029] 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 NO: 7, 12, 18, 23, 26, 34, 38, 41, 44, or 51.

[0030] 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 NO: 8, 13, 19, 24, 35, 39, 42, 45, or 52.

[0031] In some embodiments, the antibody or antigen-binding fragment thereof is selected from the group consisting of: monoclonal antibody or antigen-binding fragment thereof, polyclonal antibody or antigen-binding fragment thereof, human antibody or antigen-binding fragment thereof, humanized antibody or antigen-binding fragment thereof, primatized antibody or antigen-binding fragment thereof, bispecific antibody or antigen-binding fragment thereof, multispecific antibody or antigen-binding fragment thereof, dual variable immunoglobulin domain, monovalent antibody or antigen-binding fragment thereof, chimeric antibody or antigen-binding fragment thereof, single-chain Fv molecule (scFv), diabody, triabody, nanobody, antibody-like protein scaffold, domain antibody, Fv fragment, Fab fragment, F(ab’)2 molecule, and tandem scFv (taFv).

[0032] In some embodiments, the antibody or antigen-binding fragment thereof is a human antibody, humanized antibody, or chimeric antibody or antigen-binding fragment thereof.

[0033] In some embodiments, the antibody or antigen-binding fragment thereof specifically binds to the heterodimer of ITGA11 and integrin β1 (ITGA11B1).

[0034] In another aspect, the present disclosure provides a polynucleotide encoding any antibody or antigen-binding fragment thereof described herein.

[0035] In another aspect, the present disclosure provides a vector comprising the polynucleotide described herein (e.g., a polynucleotide encoding any antibody or antigen-binding fragment 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).

[0036] In another aspect, the present disclosure provides a host cell comprising the vector described herein. In some embodiments, the host cell expresses the antibody or antigen-binding fragment thereof described herein.

[0037] In another aspect, the present disclosure provides a pharmaceutical composition comprising the antibody or antigen-binding fragment thereof, polynucleotide, vector, or host cell described herein, and a pharmaceutically acceptable carrier or excipient.

[0038] 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, a host cell described herein, or a pharmaceutical composition described herein.

[0039] 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, a host cell described herein, or a pharmaceutical composition described herein.

[0040] In some embodiments, the disorder is a fibrotic disorder. In some embodiments, the fibrotic disorder is selected from hepatic fibrosis (e.g., fibrosis associated with cirrhosis (e.g., alcohol-induced cirrhosis, virus-induced cirrhosis, post-hepatitis C cirrhosis, and primary biliary cirrhosis), schistosomiasis, cholangitis (e.g., sclerosing cholangitis), and autoimmune-induced hepatitis), renal fibrosis (e.g., tubulointerstitial fibrosis, scleroderma, diabetic nephropathy, and glomerulonephritis), dermal fibrosis (e.g., scleroderma, hypertrophic scars, and keloids, nephrogenic fibrosing dermopathy, and burns), myelofibrosis, neurofibromatosis, fibroma, intestinal fibrosis, and surgical-induced fibrotic adhesions), cardiac fibrosis (e.g., fibrosis associated with myocardial infarction), vascular fibrosis (e.g., fibrosis associated with post-angioplasty arterial restenosis and atherosclerosis), ocular fibrosis (e.g., fibrosis associated with post-cataract surgery, proliferative vitreoretinopathy, and retro-orbital fibrosis), bone marrow fibrosis (e.g., idiopathic myelofibrosis and drug-induced myelofibrosis), pulmonary fibrosis (e.g., pulmonary interstitial fibrosis), glomerulonephritis, heart failure (ischemic and non-ischemic), scleroderma, excessive scar tissue post-surgery or device insertion, trauma or burns, progressive kidney disease, heart valve disease, hypertensive heart disease, joint and perijoint fibrosis, myelofibrosis, ocular / vitreous fibrosis, intestinal fibrosis and strictures, peritoneal and retroperitoneal fibrosis, pancreatic fibrosis, nephrogenic systemic fibrosis, primary sclerosing cholangitis, and the development of pathological matrix also plays a role in fibroproliferative tumor progression and metastasis.

[0041] In some embodiments, the disorder is an inflammatory disorder. In some embodiments, the inflammatory disorder is selected from asthma (e.g., allergic asthma, exercise-induced asthma, aspirin-sensitive / aggravated asthma, atopic asthma, severe asthma, mild asthma, moderate-to-severe asthma, asthma not previously treated with corticosteroids asthma), chronic asthma, corticosteroid-resistant asthma, corticosteroid-refractory asthma, newly diagnosed and untreated asthma, asthma caused by smoking, asthma uncontrolled by corticosteroids, etc.), airway inflammation, airway hyperreactivity, airway hyperresponsiveness, sinusitis, sinusitis with polyps, nasal polyposis, arthritis (e.g., osteoarthritis, rheumatoid arthritis, collagen-induced arthritis, arthritis caused by injury, etc.), eosinophilic inflammation, mast cell-mediated inflammatory diseases, sepsis, septic shock, seronegative enthesopathy and arthropathy (SEA) syndrome, osteoporosis, eosinophilic esophagitis, scleroderma, dermatitis, atopic dermatitis, allergic rhinitis, bullous pemphigoid, urticaria (e.g., chronic urticaria), cartilage inflammation, polymyalgia rheumatica, polyarteritis nodosa, Wegener’s granulomatosis, Behcet’s disease, myolitis, polymyolitis, dermatomyolitis, dermatomyositis, vasculitis, arteritis, diabetic nephropathy, interstitial cystitis, graft-versus-host disease (GVHD), gastrointestinal inflammatory conditions (e.g., inflammatory bowel disease (IBD), ulcerative colitis (UC), Crohn's disease (CD), colitis (e.g., colitis caused by environmental damage (e.g., caused by or associated with treatment regimens (e.g., chemotherapy, radiotherapy, etc.)), infectious colitis, ischemic colitis, collagenous or lymphocytic colitis, necrotizing enterocolitis, colitis in conditions such as chronic granulomatous disease or celiac disease, food allergy, gastritis, infectious gastritis or enterocolitis (e.g., chronic active gastritis infected with Helicobacter pylori) and other forms of gastrointestinal inflammation caused by infectious pathogens), and inflammatory lung conditions (e.g., chronic obstructive pulmonary disease (COPD), eosinophilic lung inflammation, infection-induced lung conditions (including those associated with viral (e.g., influenza, parainfluenza virus, rotavirus, human metapneumovirus, and respiratory syncytial virus), bacterial, fungal (e.g., Aspergillus), parasitic, or prion infections), allergen-induced lung conditions, pollutant-induced lung conditions (e.g., asbestosis, silicosis, or berylliosis), gastric aspiration-induced lung conditions, immune dysregulation, inflammatory conditions with a genetic predisposition (e.g., cystic fibrosis), body trauma-induced lung 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, nosocomial pneumonia, ventilator-associated pneumonia, viral pneumonia, bacterial pneumonia, and severe pneumonia), airway exacerbations, and acute respiratory distress syndrome (ARDS)).,

[0042] In some embodiments, the disorder is cancer. In some embodiments, the cancer is selected from breast cancer, colorectal cancer, hepatic cancer, renal cancer, liver cancer, lung cancer, pancreatic cancer, gastrointestinal cancer, melanoma, ovarian cancer, prostate cancer, cervical cancer, bladder cancer, glioblastoma, head and neck cancer, and cholangiocarcinoma.

[0043] Definitions

[0044] For ease of understanding the present disclosure, some terms are defined below. The terms defined herein have the meanings commonly understood by one of ordinary skill in the art relevant to the present disclosure. Terms such as "a", "an", and "the" are not intended to refer to only a single entity but include general categories that can be illustrated by specific examples. The terms herein are used to describe specific embodiments of the present disclosure, but the use of these terms does not limit the present disclosure unless outlined in the claims.

[0045] As used herein, the term "about" refers to a value that is no more than 10% higher or lower than the described value. For example, the term "about 5 nM" means a range from 4.5 nM to 5.5 nM.

[0046] As used herein, any value provided in the form of a range of values includes the upper and lower limits and any value contained within the upper and lower limits.

[0047] As used herein, the term "antibody" (Ab) refers to an immunoglobulin molecule that specifically binds or immunoreacts with a particular antigen and includes polyclonal antibodies, monoclonal antibodies, genetically engineered antibodies, and other modified forms of antibodies, including but not limited to chimeric antibodies, humanized antibodies, primatized antibodies, heteroconjugate antibodies (e.g., bispecific, trispecific, and tetra-specific antibodies, diabodies, triabodies, and tetra-bodies), and antigen-binding fragments of antibodies, including, for example, Fab', F(ab')2, Fab, Fv, rIgG, and scFv fragments. In addition, unless otherwise specified, the term "monoclonal antibody" (mAb) is intended to include the intact molecule as well as antibody fragments capable of specifically binding to the target protein (such as, for example, Fab and F(ab')2 fragments). Fab and F(ab')2 fragments lack the Fc fragment of the intact antibody, are cleared more rapidly from the animal circulation, and may have less non-specific tissue binding than the intact antibody (see Wahl et al., J. Nucl. Med. 24:316, 1983; incorporated herein by reference).

[0048] 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 fragments of the full-length antibody. Antibody fragments can be Fab, F(ab’)2, scFv, SMIP, diabodies, triabodies, affibodies, nanobodies, aptamers, bispecific antibodies, dual-binding bispecific antibodies, mAb pairs, or domain antibodies. Examples of binding fragments encompassed by the term "antigen-binding fragment" of an antibody include, but are not limited to: (i) Fab fragments, monovalent fragments consisting of the VL, VH, CL, and CH1 domains; (ii) F(ab’)2 fragments, divalent fragments containing two Fab fragments linked by disulfide bridges in the hinge region; (iii) Fd fragments consisting of the VH and CH1 domains; (iv) Fv fragments consisting of the VL and VH domains of a single arm of an antibody; (v) dAbs containing the VH and VL domains; (vi) dAb fragments consisting of the VH domain (Ward et al., Nature 341:544-546, 1989); (vii) dAbs consisting of the VH or VL domain; (viii) isolated complementarity-determining regions (CDRs); and (ix) combinations of two or more isolated CDRs, which may optionally be linked by a synthetic linker. In addition, although the two domains, VL and VH, of an Fv fragment are encoded by separate genes, they can be joined using recombinant methods by a linker such that they can be prepared as a single protein chain, wherein the VL and VH regions pair to form a monovalent molecule (called 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 procedures known in the art.

[0049] As used herein, the term "binding affinity" refers to the strength of the total 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 stated, as used herein, "binding affinity" refers to the intrinsic binding affinity, which reflects the specific interactions between the 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 contain antibodies that generally tend to dissociate readily from the antigen, while high-affinity complexes contain antibodies that generally tend to remain bound to the antigen for a longer period of time.

[0050] As used herein, the term "chimeric" antibody refers to an antibody having variable domain sequences (e.g., CDR sequences) of immunoglobulin derived from one source organism (e.g., rat or mouse), and constant regions of immunoglobulin derived from a different organism (e.g., human, another primate, pig, goat, rabbit, hamster, cat, dog, guinea pig, members of the bovine family (e.g., cattle, bison, buffalo, elk, and yaks, etc.), cow, sheep, horse, or bison, etc.). Methods for producing 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; which are incorporated herein by reference.

[0051] As used herein, the term "complementary determining region" (CDR) refers to the hypervariable regions present in the variable domains of the light and heavy chains. The more conserved portions of the variable domains are called framework regions (FR). As understood in the art, the amino acid positions that delineate the hypervariable regions of an antibody can vary, depending on context and the various definitions known in the art. Some positions within the variable domain can be considered mixed hypervariable positions, as these positions can be considered to be within the hypervariable region under one set of criteria and outside the hypervariable region under another set of criteria. One or more of these positions can also be present in an extended hypervariable region. The antibodies described herein can include modifications of these mixed hypervariable positions. The variable domains of the native heavy and light chains each contain four framework regions that predominantly adopt a β-sheet conformation, which are connected by three CDRs that form loops connecting the β-sheet structures and, in some cases, form part of the β-sheet structure. The CDRs in each chain are held together in close proximity in the order FR1-CDR1-FR2-CDR2-FR3-CDR3-FR4 by the FR regions and, together with the CDRs from the other antibody chain, contribute to the formation of the target binding site of the antibody (see Kabat et al, Sequences of Proteins of Immunological Interest (National Institute of Health, Bethesda, Md. 1987; incorporated herein by reference). As used herein, unless otherwise specified, the numbering of immunoglobulin amino acid residues is according to the Kabat et al immunoglobulin amino acid residue numbering system.

[0052] As used herein, the terms "conservative mutation", "conservative substitution" or "conservative amino acid substitution" refer to the substitution of one or more amino acids with one or more different amino acids that exhibit similar physicochemical properties (e.g., polarity, net charge, and steric volume). These properties of the twenty naturally occurring amino acids are summarized in Table 1 below.

[0053] Table 1. Representative physicochemical properties of the naturally occurring amino acids

[0054]

[0055] As can be seen from the table, the 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. Thus, a conservative mutation or substitution is a mutation or substitution that replaces one amino acid with a member of the same amino acid family (e.g., substituting Ser for Thr, or Lys for Arg).

[0056] Amino acid substitutions can be represented herein using the following convention: (AA1)(N)(AA2), where "AA1" represents the amino acid that is normally present at a specific position in an amino acid sequence, "N" represents the residue number at which the substitution occurs in the amino acid sequence, and "AA2" represents the amino acid that is present in the amino acid sequence after the substitution is complete. For example, in the context of an antibody hinge region (e.g., the IgG2 antibody hinge region), the symbol "C232S" refers to the substitution of the naturally occurring cysteine residue with a serine residue at amino acid residue 232 of the indicated hinge amino acid sequence. Similarly, in the context of an antibody hinge region (e.g., the IgG2 antibody hinge region), the symbol "C233S" refers to the substitution of the naturally occurring cysteine residue with a serine residue at amino acid residue 233 of the indicated hinge amino acid sequence.

[0057] As used herein, the term "conjugate" refers to a compound formed by the chemical bonding of a reactive functional group of one molecule with an appropriate reactive functional group of another molecule.

[0058] As used herein, the term "derived antibody" refers to an antibody that has been modified by a chemical reaction to cleave residues or add chemical moieties that are not native to the isolated antibody. Derived antibodies can be obtained by glycosylation, acetylation, polyethylene glycolylation, phosphorylation, amidation, derivatization by addition of known chemical protecting / blocking groups, proteolytic cleavage, or conjugation to a cell ligand or other protein. Any of the various chemical modifications can be carried out using established procedures by known techniques, including but not limited to specific chemical cleavage, acetylation, formylation, metabolic synthesis with tunicamycin, etc. In addition, derivatives can contain one or more non-natural amino acids, for example, using amber suppression techniques (see, e.g., U.S. Patent No. 6,964,859; incorporated herein by reference).

[0059] As used herein, the term "diabody" refers to a bivalent antibody comprising two polypeptide chains, where each polypeptide chain comprises V H and V LA domain, the linker is too short (e.g., a linker containing five amino acids) to allow intramolecular association of the VH and VL domains on the same polypeptide chain. This configuration forces each domain to pair with the complementary domain on another polypeptide chain, thus forming a homodimer structure. Correspondingly, the term "trimeric antibody" refers to a trivalent antibody containing three polypeptide chains, each polypeptide chain containing a VH domain and a VL domain linked by a linker that is extremely short (e.g., a linker containing 1-2 amino acids) to not allow intramolecular association of the VH and VL domains within the same polypeptide chain. In order to fold into their native structure, peptides configured in this way are usually trimerized, thereby placing the VH and VL domains of adjacent polypeptide chains in spatial proximity to each other to allow proper folding (see Holliger et al., Proc. Natl. Acad. Sci. USA 90:6444-48, 1993; incorporated herein by reference).

[0060] As used herein, the term "disorder" refers to any condition, disease, or pathogenic abnormal biological function state of a subject. In particular, the present disclosure provides disorders related to ITGA11. Specific disorders of the present disclosure include fibrotic disorders, inflammatory disorders, or cancers.

[0061] As used herein, the term "epitope" refers to the antigenic portion recognized and bound by a polypeptide (e.g., an antibody, an antigen-binding fragment thereof, a single-chain polypeptide, or a construct described herein). In the context of a protein antigen (e.g., ITGA11), an epitope can 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 constituent amino acids bind to a polypeptide (e.g., an antibody, an antigen-binding fragment thereof, a single-chain polypeptide, or a construct thereof). A continuous epitope can contain, for example, 1, 5, 10, 15, 20 or more amino acids within the antigen. In some embodiments, an epitope can be a discontinuous epitope, which contains two or more amino acid segments, each segment being separated from each other by one or more intervening amino acid residues in the amino acid sequence of the antigen. A discontinuous epitope can contain, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more such amino acid residue segments. Although separated by intervening amino acids, the segments constituting a discontinuous epitope can, for example, be in spatial proximity to each other in the three-dimensional conformation of the antigen. An epitope can be defined not only by its amino acid composition, but also by the post-translational state of the epitope amino acids (e.g., phosphorylation) or the bond geometry (e.g., cis or trans) of the peptide bond between two amino acids in the epitope.

[0062] As used herein, the term "framework region" or "FW region" encompasses the amino acid residues adjacent to the CDRs. FW region residues can 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.

[0063] As used herein, the term "fusion protein" refers to a protein that is linked to another molecule by a covalent bond. A fusion protein can be chemically synthesized, for example, by a reaction to form an amide bond between the N-terminus of one protein and the C-terminus of another protein. Optionally, a fusion protein comprising one protein covalently bound to another protein can be recombinantly expressed in a cell (e.g., a eukaryotic or prokaryotic cell) by expressing a polynucleotide encoding the fusion protein from a vector or the genome of the cell. A fusion protein can contain one protein covalently bound to a linker, which in turn is covalently bound to another molecule. Examples of linkers that can be used to form fusion proteins include peptide-containing linkers, such as linkers containing naturally occurring or non-naturally occurring amino acids. In some embodiments, it may be desirable to include D-amino acids in the linker because these residues are not present in naturally occurring proteins and are thus more resistant to degradation by endogenous proteases. Linkers can be prepared using a variety of strategies well known in the art and can be cleaved by enzymatic hydrolysis, photolysis, hydrolysis under acidic conditions, hydrolysis under alkaline conditions, oxidation, disulfide reduction, nucleophilic cleavage, or organometallic cleavage, depending on the reactive components of the linker (Leriche et al., Bioorg. Med. Chem., 20:571-582, 2012).

[0064] As used herein, the term "heterospecific antibody" refers to a monoclonal antibody that has binding specificity for at least two different antigens, preferably a human antibody or a humanized antibody. Traditionally, the recombinant production of heterospecific antibodies has been based on the co-expression of two immunoglobulin heavy chain-light chain pairs, where the two heavy chains have different specificities (Milstein et al., Nature 305:537, 1983). Similar procedures are disclosed, 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; 5,932,448; 5,833,985; 5,821,333; 5,807,706; 5,643,759; 5,601,819; 5,582,996; 5,496,549; 4,676,980, WO 91 / 00360, WO 92 / 00373, EP 03089, Traunecker et al., EMBO J. 10:3655 (1991), Suresh et al., Methods in Enzymology 121:210 (1986); which are incorporated herein by reference. Heterospecific antibodies can include Fc mutations that enforce correct chain association in multispecific antibodies, as described in Klein et al, mAbs 4(6):653-663, 2012; which is incorporated herein by reference.

[0065] As used herein, the term "human antibody" refers to an antibody in which substantially every part of the protein (e.g., CDRs, frameworks, C L 、C H structural domains (e.g., C H 1, C H 2, C H 3), hinge, (V L 、V H)) are substantially non-immunogenic in humans and have only minor sequence changes or variations. Human antibodies can be produced in human cells (e.g., by recombinant expression), or by non-human animals or prokaryotic or eukaryotic cells capable of expressing functional rearranged human immunoglobulin (e.g., heavy and / or light chain) genes. In addition, when a human antibody is a single-chain antibody, it can contain a linker peptide not present in native human antibodies. For example, an Fv can contain a linker peptide (e.g., two to about eight glycine or other amino acid residues) that connects the variable region of the heavy chain and the variable region of the light chain. Such linker peptides are considered to be of human origin. Human antibodies can be prepared by a variety of methods known in the art, including phage display methods using antibody libraries derived from human immunoglobulin sequences. See U.S. Patent Nos. 4,444,887 and 4,716,111; and PCT Publications WO 1998 / 46645; WO 1998 / 50433; WO1998 / 24893; WO 1998 / 16654; WO 1996 / 34096; WO 1996 / 33735; and WO 1991 / 10741, which are incorporated herein by reference. Human antibodies can also be produced using transgenic mice that are unable to express functional endogenous immunoglobulins but can express human immunoglobulin genes. See, e.g., PCT Publications WO 98 / 24893, WO 92 / 01047, WO 96 / 34096, WO 96 / 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, which are incorporated herein by reference.

[0066] As used herein, the term "humanized" antibody refers to a form of a non-human (e.g., murine) antibody that is a chimeric immunoglobulin, immunoglobulin chain, or fragment thereof (e.g., Fv, Fab, Fab', F(ab')2 or other target-binding subdomains of an antibody) that contains a minimal sequence derived from a non-human immunoglobulin. Generally, a humanized antibody will contain substantially all of at least one and usually two variable domains, wherein all or substantially all of the CDR regions correspond to the CDR regions of a non-human immunoglobulin. All or substantially all of the FR regions may also be the FR regions of a human immunoglobulin sequence. A humanized antibody may also contain at least a portion of an immunoglobulin constant region (Fc), typically a human immunoglobulin consensus sequence. Methods for humanizing antibodies are known in the art. See, e.g., Riechmann et al., Nature 332:323-7, 1988; U.S. Patents to Queen et al.: Nos. 5,530,101; 5,585,089; 5,693,761; 5,693,762; and 6,180,370; EP239400; PCT Publication WO 91 / 09967; U.S. Patent No. 5,225,539; EP592106; and EP519596; which are incorporated herein by reference.

[0067] As used herein, the term "hydrophobic side chain" refers to an amino acid side chain that exhibits relatively low solubility in water due to, for example, the steric or electronic properties of chemical moieties present within the side chain. Examples of amino acids containing hydrophobic side chains include amino acids containing unsaturated aliphatic hydrocarbons, such as alanine, valine, leucine, isoleucine, proline, and methionine, and amino acids containing aromatic ring systems that are electrostatically neutral at physiological pH, such as tryptophan, phenylalanine, and tyrosine.

[0068] As used herein, the term "monoclonal antibody" refers to an antibody derived from a single clone, including any eukaryotic, prokaryotic, or phage clone, regardless of the method by which it is produced.

[0069] As used herein, the term "non-natural constant region" refers to an antibody constant region that is derived from a source different from the antibody variable region, or is an antibody constant region of a synthetic polypeptide produced by a human having an amino acid sequence different from that of a natural antibody constant region. For example, an antibody containing a non-natural 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 (e.g., cow, bison, buffalo, elk, and yak, etc.), cow, sheep, horse, or bison, etc.).

[0070] 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 the amino acid (or nucleic acid) residues of a reference sequence after aligning the sequences and introducing gaps, if necessary, to achieve the maximum percent sequence identity (e.g., gaps may be introduced in one or both of the candidate and reference sequences to achieve optimal alignment, and non-homologous sequences may be disregarded for comparison purposes). The alignment for purposes of determining percent sequence identity can be achieved in various ways within the skill in the art, e.g., using publicly available computer software such as BLAST, ALIGN, or Megalign (DNASTAR) software. Those skilled in the art can determine appropriate parameters for measuring the alignment, including any algorithms required to achieve maximum alignment over the full length of the sequences being compared. For example, a reference sequence aligned for comparison with a candidate sequence can show that the candidate sequence exhibits 50% to 100% sequence identity over the full length of the candidate sequence or over a selected contiguous portion of the amino acid (or nucleic acid) residues of the candidate sequence. The length of a candidate sequence aligned for comparison purposes can be, for example, at least 30% of the length of the reference sequence (e.g., 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100%). A molecule is identical at a position when the position in the candidate sequence is occupied by the same amino acid residue as the corresponding position in the reference sequence.

[0071] As used herein, the term "primateized antibody" refers to an antibody that contains framework regions from an antibody of primate origin and other regions (e.g., CDRs and / or constant regions) from an antibody of non-primate origin. Methods for producing primateized antibodies are known in the art. See, e.g., U.S. Patent Nos. 5,658,570; 5,681,722; and 5,693,780, which are incorporated herein by reference. For example, the primateized antibodies or antigen-binding fragments 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 containing one or more primate framework regions.

[0072] As used herein, the term "operably linked" in the context of 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.

[0073] As used herein, the term "pharmacokinetic profile" refers to the absorption, distribution, metabolism, and clearance of a drug over time after administration of the drug to a patient.

[0074] 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, 1990); which is incorporated herein by reference.

[0075] As used herein, the term "scFv" refers to a single-chain Fv antibody in which the variable domains from the heavy and light chains of an antibody are joined to form a single chain. The scFv fragment contains a single polypeptide chain that comprises an antibody light chain variable region (VL) (e.g., CDR-L1, CDR-L2, and / or CDR-L3) and an antibody heavy chain variable region (VH) (e.g., CDR-H1, CDR-H2, and / or CDR-H3) separated by a linker. The linker that joins the VL and VH regions of the scFv fragment can be a peptide linker that comprises proteogenic amino acids. Alternative linkers can be used to increase the resistance of the scFv fragment to proteolytic degradation (e.g., linkers containing D-amino acids), to improve the solubility of the scFv fragment (e.g., hydrophilic linkers such as linkers containing polyethylene glycol or polypeptides containing repeating 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 attenuate 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 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 the scFv molecule can be derived from one or more antibody molecules. Those of ordinary skill in the art will also understand that the variable regions of the scFv molecules described herein can be modified such that their amino acid sequences are different from those of the antibody molecules from which they are derived. For example, in one embodiment, nucleotide or amino acid substitutions can be made that result in conservative substitutions or alterations at amino acid residues (e.g., in CDRs and / or framework residues). Optionally, or alternatively, CDR amino acid residues are mutated using techniques well known in the art to optimize antigen binding. scFv fragments are described, for example, in WO 2011 / 084714; which is incorporated herein by reference.

[0076] As used herein, the phrase "specifically binds" refers to a binding reaction that determines the presence of an antigen in a heterogeneous population of proteins and other biomolecules, the antigen being specifically recognized, for example, by an antibody or an antigen-binding fragment thereof. An antibody or an antigen-binding fragment thereof that specifically binds to an antigen will have a K of less than 100 nM DBinds to an antigen. For example, an antibody or antigen-binding fragment thereof that specifically binds to an antigen will have a K of no more than 100 nM (e.g., 1 pM to 100 nM). D Binds to an antigen. An antibody or antigen-binding fragment thereof that does not exhibit specific binding to a particular antigen or epitope thereof will have a K greater than 100 nM (e.g., greater than 500 nm, 1 μM, 100 μM, 500 μM, or 1 mM) for that particular antigen or epitope thereof. D A variety of immunoassay formats can be used to select antibodies that specifically immunoreact with a particular protein or carbohydrate. For example, solid-phase ELISA immunoassays are commonly used to select antibodies that specifically immunoreact 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).

[0077] As used herein, the terms "subject" and "patient" refer to an organism that is undergoing treatment for a particular disease or condition as described herein. Examples of subjects and patients include mammals undergoing treatment for a disease or condition, such as humans, primates, pigs, goats, rabbits, hamsters, cats, dogs, guinea pigs, members of the bovine family (e.g., cows, bison, buffalo, elk, and yaks, etc.), cows, sheep, horses, and bison, etc.

[0078] As used herein, the term "transfection" refers to any of a variety of techniques commonly used to introduce exogenous DNA into a prokaryotic or eukaryotic host cell, such as electroporation, lipofection, calcium phosphate precipitation, DEAE-dextran transfection, etc.

[0079] As used herein, the term "treat" or "treatment" refers to therapeutic treatment, wherein the aim is to prevent or slow down (reduce) an undesired physiological change or disorder, such as the progression of a fibrotic disorder, an inflammatory disorder, or cancer as described herein. Beneficial or desired clinical outcomes include, but are not limited to, alleviation of symptoms, reduction in the degree of the disease, stabilization of the disease state (i.e., not getting worse), delay or slowing of disease progression, improvement or amelioration of the disease state, and remission (whether partial or complete), whether detectable or not. Subjects in need of treatment include subjects who already have a condition or disorder, as well as subjects who are predisposed to having a condition or disorder, or subjects in need of preventing a condition or disorder.

[0080] As used herein, the term "variable region CDR" encompasses the amino acids in the CDRs or complementarity determining regions identified according to sequence- or structure-based methods of use. As used herein, the term "CDR" or "complementarity determining region" refers to the discontinuous antigen-binding sites present within the variable regions of heavy and light chain polypeptides. These specific regions have been described by Kabat et al., J. Biol. Chem. 252:6609-6616, 1977 and Kabat, et al., Sequences of Proteins of Immunological Interest, Fifth 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), where the definitions include overlapping or subsets of amino acid residues when compared to one another. The term "CDR" can be, for example, the CDR defined by Kabat based on sequence comparison.

[0081] As used herein, the term "vector" includes nucleic acid vectors, e.g., DNA vectors such as plasmids, RNA vectors, viruses or other suitable replicons (e.g., viral vectors). A variety of vectors have been developed for delivering polynucleotides encoding foreign proteins into prokaryotic or eukaryotic cells. Examples of such expression vectors are disclosed, for example, in WO 1994 / 11026; which is incorporated herein by reference. The expression vectors described herein contain polynucleotide sequences and additional sequence elements, e.g., for expressing proteins and / or integrating these polynucleotide sequences into the genome of mammalian cells. Certain vectors that can be used to express the antibodies and antibody fragments described herein include plasmids containing regulatory sequences (e.g., promoter and enhancer regions that direct gene transcription). Other useful vectors for expressing antibodies and antibody fragments contain polynucleotide sequences that increase the translation rate of these genes or improve the stability or nuclear export of the mRNA produced by gene transcription. These sequence elements include, for example, 5' and 3' untranslated regions, internal ribosome entry sites (IRES), and polyadenylation signal sites to direct efficient transcription of the genes carried on the expression vectors. The expression vectors described herein may also contain polynucleotides encoding markers for selecting cells containing the vector. Examples of suitable markers include genes encoding antibiotic (e.g., ampicillin, chloramphenicol, kanamycin or nourseothricin) resistance.

[0082] As used herein, the term "VH" refers to the variable region of the 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 the immunoglobulin light chain, including the light chain of an Fv, scFv, dsFv or Fab. Antibodies (Abs) and immunoglobulins (Igs) are glycoproteins with the same structural features. Antibodies exhibit binding specificity for a particular target, while immunoglobulins include antibodies and other antibody-like molecules lacking target specificity. Native antibodies and immunoglobulins are typically heterotetrameric glycoproteins of about 150,000 daltons, composed of two identical light chains (L) and two identical heavy chains (H). Each heavy chain of a native antibody has a variable domain (VH) at the amino terminus, followed by a number of constant domains. Each light chain of a native antibody has a variable domain at the amino terminus (VL) and a constant domain at the carboxyl terminus. Brief Description of the Drawings

[0084] Figure 1 Shows that anti-ITGA11 antibodies FIB-918-1, FIB-918-2, FIB-918-3 and FIB-918-4 inhibit the binding of collagen to human ITGA11 expressed in mouse C2C12 cells.

[0085] Figure 2 Shows that anti-ITGA11 antibodies FIB-918-5, FIB-918-6, FIB-918-7 and FIB-918-8 inhibit the binding of collagen to human ITGA11 expressed in mouse C2C12 cells.

[0086] Figure 3 Shows that anti-ITGA11 antibodies FIB-918-9 and FIB-918-10 inhibit the binding of collagen to human ITGA11 expressed in mouse C2C12 cells.

[0087] Detailed Description

[0088] The present disclosure provides antibodies or antigen-binding fragments thereof that specifically bind to integrin α11 (ITGA11). The present disclosure also provides methods for treating ITGA11-related disorders (e.g., fibrotic disorders, inflammatory disorders or cancers) in a subject. The present disclosure also includes related pharmaceutical compositions, polynucleotides, vectors, host cells, preparation methods, treatment methods, diagnostic methods and kits.

[0089] I. Anti-ITGA11 Antibodies or Antigen-Binding Fragments Thereof

[0090] The present disclosure provides an antibody or an antigen-binding fragment thereof that specifically binds to integrin chain α11 (ITGA11). In some embodiments, the antibody or antigen-binding fragment thereof disclosed herein specifically binds to human ITGA11, cynomolgus monkey ITGA11, and / or mouse ITGA11. In some embodiments, the antibody or antigen-binding fragment thereof disclosed herein specifically binds to ITGA11 (α11β1 or ITGA11B1) that heterodimerizes with integrin chain β1 (ITGB1) (e.g., human ITGA11B1, cynomolgus monkey ITGA11B1, or mouse ITGA11B1). In some embodiments, the antibody or antigen-binding fragment thereof disclosed herein specifically binds to human ITGA11 that heterodimerizes with human ITGB1, cynomolgus monkey ITGB1, or mouse ITGB1.

[0091] In some embodiments, the antibody or antigen-binding fragment thereof disclosed herein specifically binds to ITGA11 (e.g., human ITGA11) relative to ITGA10 (e.g., human ITGA10) or ITGA2 (e.g., human ITGA2). In some embodiments, the antibody or antigen-binding fragment thereof disclosed herein specifically binds to ITGA11B1 (e.g., human ITGA11B1) relative to ITGA10B1 (e.g., human ITGA10B1) or ITGA2B1 (e.g., human ITGA2B1).

[0092] In some embodiments, the antibody or antigen-binding fragment thereof disclosed herein is a monovalent, bispecific antibody.

[0093] In some embodiments, the antibody or antigen-binding fragment thereof disclosed herein preferentially binds ITGA11 and ITGA10 compared to ITGA2 or ITGB1.

[0094] In particular, the present disclosure features an anti-ITGA11 antibody or antigen-binding fragment described according to 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 heavy chain pairings comprising any heavy chain variable region (VH) described herein paired with a light chain pairing comprising any light chain variable region (VL) disclosed herein.

[0095] mAb FIB-918-1

[0096] mAb FIB-918-1 contains complementarity-determining region (CDR)-heavy chain 1 (CDR-H1) containing the amino acid sequence GFTFSSYA (SEQ ID NO:1); CDR-heavy chain 2 (CDR-H2) containing the amino acid sequence ISGSGGST (SEQ ID NO:2); and CDR-heavy chain 3 (CDR-H3) containing the amino acid sequence AKDLDWSGHDAFDI (SEQ ID NO:3).

[0097] mAb FIB-918-1 contains a heavy chain variable domain having the amino acid sequence EVQLVESGGGLVQPGGSLRLSCAAS GFTFSSYA MSWVRQAPGKGLEWVSA ISGSGGST YYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYC AKDLDWSGHDAFDI WGQGTTVTVSS (SEQ ID NO:7; CDRs underlined).

[0098] mAb FIB-918-1 contains complementarity-determining region (CDR)-light chain 1 (CDR-L1) containing the amino acid sequence QSISSY (SEQ ID NO:4); CDR-light chain 2 (CDR-L2) containing the amino acid sequence AAS (SEQ ID NO:5); and CDR-light chain 3 (CDR-L3) containing the amino acid sequence QQTYSTPLT (SEQ ID NO:6).

[0099] mAb FIB-918-1 contains a light chain variable domain having the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRAS QSISSY LNWYQQKPGKAPKLLIY AAS SLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYC QQTYSTPLT FGGGTKVEIKR (SEQ ID NO:8; CDRs underlined).

[0100] mAb FIB-918-2

[0101] mAb FIB-918-2 contains CDR-H1 containing the amino acid sequence GFTFSSYA (SEQ ID NO:1); CDR-H2 containing the amino acid sequence ISGSGGST (SEQ ID NO:2); and CDR-H3 containing the amino acid sequence AKDPRGSGRDDAFDI (SEQ ID NO:20).

[0102] mAb FIB-918-2 comprises a heavy chain variable domain having the amino acid sequence EVQLLESGGGLVQPGGSLRLSCAAS GFTFSSYA MSWVRQAPGKGLEWVSA ISGSGGST YYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYD AKDPRGSGRDDAFDI WGQGTMVTVSS (SEQ ID NO: 23; CDRs underlined).

[0103] mAb FIB-918-2 comprises CDR-L1 containing the amino acid sequence QTIGSY (SEQ ID NO: 21); CDR-L2 containing the amino acid sequence GAS (SEQ ID NO: 22); and CDR-L3 containing the amino acid sequence QQSYSTPFT (SEQ ID NO: 11).

[0104] mAb FIB-918-2 comprises a light chain variable domain having the amino acid sequence NIQMTQSPSSLSASVGDRVTITCRAS QTIGSY LNWYQQKPGTAPKLLIY GAS TSHTWVPSRFTGGGSGTEFTLTISSLQSEDFATYYC QQSYSTPFT FGGGTKLEIKR (SEQ ID NO: 24; CDRs underlined).

[0105] mAb FIB-918-3

[0106] mAb FIB-918-3 comprises CDR-H1 containing the amino acid sequence GFTFSSYS (SEQ ID NO: 14); CDR-H2 containing the amino acid sequence ISSSSSTI (SEQ ID NO: 15); and CDR-H3 containing the amino acid sequence ARGPDLSDYFDY (SEQ ID NO: 16).

[0107] mAb FIB-918-3 comprises a heavy chain variable domain having the amino acid sequence EVQLVESGGGLVQPGGSLRLSCAAS GFTFSSYS MNWVRQAPGKGLEWVSY ISSSSSTI YYADSVKGRFTISRDNAKNSLYLQMNsLRAEDTAVYYC ARGPDLSDYFDY WGRGTLVTVSS (SEQ ID NO: 18; CDRs underlined).

[0108] mAb FIB-918-3 comprises a CDR-L1 containing the amino acid sequence QSISSY (SEQ ID NO: 4); a CDR-L2 containing the amino acid sequence AAS (SEQ ID NO: 5); and a CDR-L3 containing the amino acid sequence QQSYSTPLT (SEQ ID NO: 17).

[0109] mAb FIB-918-3 comprises a light chain variable domain having the amino acid sequence DIQLTQSPSSLSASVGDRVTITCRAS QSISSY LNWYQQKPGKAPKLLIY AAS SLQSGVPSRFSGSGSGTDFTLTIssLQPEDIATYYC QQSYSTPL TFGGGTKLEIKR (SEQID NO: 19; CDRs underlined).

[0110] mAb FIB-918-4

[0111] mAb FIB-918-4 comprises a CDR-H1 containing the amino acid sequence GYTFTDYY (SEQ ID NO: 28); a CDR-H2 containing the amino acid sequence FDPEDGET (SEQ ID NO: 29); and a CDR-H3 containing the amino acid sequence ATLDYRGVVYFDY (SEQ IDNO: 30).

[0112] mAb FIB-918-4 comprises a heavy chain variable domain having the amino acid sequence QVQLVQSGAEVKKPGATVKISCKVS GYTFTDYY MHWVRQAPGKGLEWMGG FDPEDGET IYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYC ATLDYRGVVYFDY WGQGTLVTVSS (SEQ ID NO: 34; CDRs underlined).

[0113] mAb FIB-918-4 comprises a CDR-L1 containing the amino acid sequence SGSIASNY (SEQ ID NO: 31); a CDR-L2 containing the amino acid sequence EDK (SEQ ID NO: 32); and a CDR-L3 containing the amino acid sequence QSYDSSNHWV (SEQ ID NO: 33).

[0114] mAb FIB-918-4 comprises a light chain variable domain having the amino acid sequence NFMLTQPHSVSDSPGKTVTISCTGS SGSIASNY VQWYQQRPGSAPTTVIY EDKRRPSGVPDRFIGSIDSSSNSASLTISGLRTEDEADYYC QSYDSSNHWV The light chain variable domain of FGGGTQLTVLG (SEQ ID NO: 35; CDR underlined).

[0115] mAb FIB-918-5

[0116] mAb FIB-918-5 comprises CDR-H1 containing the amino acid sequence GFTFSSYA (SEQ ID NO: 1); CDR-H2 containing the amino acid sequence ISGSGGST (SEQ ID NO: 2); and CDR-H3 containing the amino acid sequence AKDPTTMTTDAFDI (SEQ ID NO: 25).

[0117] mAb FIB-918-5 comprises having the amino acid sequence EVQLVESGGGLVQPGGSLRLSCAAS GFTFSSYA MSWVRQAPGKGLEWVSA ISGSGGST YYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYC AKDPTTMTTDAFDI The heavy chain variable domain of WGQGTMVTVSS (SEQ ID NO: 26; CDR underlined).

[0118] mAb FIB-918-5 comprises CDR-L1 containing the amino acid sequence QSISSY (SEQ ID NO: 4); CDR-L2 containing the amino acid sequence AAS (SEQ ID NO: 5); and CDR-L3 containing the amino acid sequence QQSYSTPFT (SEQ ID NO: 11).

[0119] mAb FIB-918-5 comprises having the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRAS QSISSY LNWYQQKPGKAPKLLIY AAS SLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYC QQSYSTPFT The light chain variable domain of FGPGTKLEIKR (SEQID NO: 27; CDR underlined).

[0120] mAb FIB-918-6

[0121] mAb FIB-918-6 comprises CDR-H1 containing the amino acid sequence GFTFSNAW (SEQ ID NO: 9); CDR-H2 containing the amino acid sequence ISGSGGST (SEQ ID NO: 2); and CDR-H3 containing the amino acid sequence ARDRGYSYSETSNDAFDI (SEQ ID NO: 10).

[0122] mAb FIB-918-6 comprises a heavy chain variable domain having the amino acid sequence EVQLLESGGDLVKAGGSLRLSCAAS GFTFSNAW MSWVRQAPGKGLEWVSG ISGSGGST YYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYC ARDRGYSYSETSNDAFD I WGRGTLVTVSS (SEQ ID NO: 12; CDRs underlined).

[0123] mAb FIB-918-6 comprises CDR-L1 containing the amino acid sequence QSISSY (SEQ ID NO: 4); CDR-L2 containing the amino acid sequence AAS (SEQ ID NO: 5); and CDR-L3 containing the amino acid sequence QQSYSTPFT (SEQ ID NO: 11).

[0124] mAb FIB-918-6 comprises a light chain variable domain having the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRAS QSISSY LNWYQQKPGKAPKLLIY AAS SLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQ QSYSTPFT FGPGTKLEIKR (SEQ ID NO: 13; CDRs underlined).

[0125] mAb FIB-918-7

[0126] mAb FIB-918-7 comprises CDR-H1 containing the amino acid sequence GFMFDTHA (SEQ ID NO: 46); CDR-H2 containing the amino acid sequence ISGSGGSI (SEQ ID NO: 47); and CDR-H3 containing the amino acid sequence ARSGETAGTDYFDY (SEQ ID NO: 48).

[0127] mAb FIB-918-7 comprises a heavy chain variable domain having the amino acid sequence EVQLVESGGGLVQPGGSLRLSCAAS GFMFDTHAMSWVRQAPGKGLEWVSS ISGSGGSI YYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYC ARSGETAGTDYFDY The heavy chain variable domain of WGQGTLVTVSS (SEQ ID NO: 51; CDRs underlined).

[0128] mAb FIB-918-7 comprises a CDR-L1 containing the amino acid sequence QSVSSSY (SEQ ID NO: 49); a CDR-L2 containing the amino acid sequence GAS (SEQ ID NO: 22); and a CDR-L3 containing the amino acid sequence QQDYNSPYT (SEQ ID NO: 50).

[0129] mAb FIB-918-7 comprises having the amino acid sequence EIVMTQSPATLSLSPGERATLSCRAS QSVSSSY LSWYQQKPGQAPRLLIY GAS TRATGIPARFSGSGSGTDFTLTISSLQPEDFAVYYC QQDYNSPYT The light chain variable domain of FGQGTKVDIKR (SEQID NO: 52; CDRs underlined).

[0130] mAb FIB-918-8

[0131] mAb FIB-918-8 comprises a CDR-H1 containing the amino acid sequence GFTFSNAW (SEQ ID NO: 9); a CDR-H2 containing the amino acid sequence ISGSGGST (SEQ ID NO: 2); and a CDR-H3 containing the amino acid sequence AKQTVTSADDYFDY (SEQ ID NO: 43).

[0132] mAb FIB-918-8 comprises having the amino acid sequence EVQLVESGGGWRPGGPLRLSCAAS GFTFSNAW MSWVRQAPGKGLEWVSA ISGSGGST YYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAIYYC AKQTVTSADDYFDY The heavy chain variable domain of WGQGTLVTVSS (SEQ ID NO: 44; CDRs underlined).

[0133] mAb FIB-918-8 contains CDR-L1 with the amino acid sequence QSISSY (SEQ ID NO: 4); CDR-L2 with the amino acid sequence AAS (SEQ ID NO: 5); and CDR-L3 with the amino acid sequence QQSYSTPFT (SEQ ID NO: 11).

[0134] mAb FIB-918-8 contains a light chain variable domain having the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRAS QSISSY LNWYQQKPGKAPKLLIY AAS SLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYC QQSYSTPFT FGPGTKLEIKR (SEQ ID NO: 45; CDRs underlined).

[0135] mAb FIB-918-9

[0136] mAb FIB-918-9 contains CDR-H1 with the amino acid sequence GFTFSDYW (SEQ ID NO: 36); CDR-H2 with the amino acid sequence ISGSGGST (SEQ ID NO: 2); and CDR-H3 with the amino acid sequence AKDLLWAARDAFDI (SEQ ID NO: 37).

[0137] mAb FIB-918-9 contains a heavy chain variable domain having the amino acid sequence EVQLVESGGGLVQPGGSLRLSCLAS GFTFSDYW MAWVRQAPGKGLEWVSA ISGSGGST YYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYC AKDLLWAARDAFDI WGQGTLVTVSS (SEQ ID NO: 38; CDRs underlined).

[0138] mAb FIB-918-9 contains CDR-L1 with the amino acid sequence QSISSY (SEQ ID NO: 4); CDR-L2 with the amino acid sequence AAS (SEQ ID NO: 5); and CDR-L3 with the amino acid sequence QQSYSTPFT (SEQ ID NO: 11).

[0139] mAb FIB-918-9 contains a light chain variable domain having the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRAS QSISSY LNWYQQKPGKAPKLLIY AASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYC QQSYSTPFT The light chain variable domain of FGPGTKLEIKR (SEQ ID NO: 39; CDR underlined).

[0140] mAb FIB-918-10

[0141] mAb FIB-918-10 comprises CDR-H1 containing the amino acid sequence GFTFSSYA (SEQ ID NO: 1); CDR-H2 containing the amino acid sequence ISGSGGST (SEQ ID NO: 2); and CDR-H3 containing the amino acid sequence AKDLLWAARDAFDI (SEQ ID NO: 37).

[0142] mAb FIB-918-10 comprises the amino acid sequence QVQLVESGGGLVQPGGSLRLSCAAS GFTFSSYA MSWVRQAPGKGLEWVSA ISGSGGST YYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYC AKDLLWAARDAFDI The heavy chain variable domain of WGQGTLVTVSS (SEQ ID NO: 41; CDR underlined).

[0143] mAb FIB-918-10 comprises CDR-L1 containing the amino acid sequence QGINDF (SEQ ID NO: 40); CDR-L2 containing the amino acid sequence AAS (SEQ ID NO: 5); and CDR-L3 containing the amino acid sequence QQSYSTPLT (SEQ ID NO: 17).

[0144] mAb FIB-918-10 comprises the amino acid sequence AlQLTQSPSTLSASVGDRVTITCRAS QGINDF LAWYQQKPGKAPKLLIY AAS SLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYC QQSYSTPLT The light chain variable domain of FGGGTKVEIKR (SEQ ID NO: 42; CDR underlined).

[0145] Light chain variable domain sequence

[0146] The present disclosure provides an antibody or an antigen-binding fragment thereof that comprises a light chain variable domain that comprises 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 NOs: 8, 13, 19, 24, 35, 39, 42, 45 and 52.

[0147] SEQ ID NO: 8

[0148]

[0149] SEQ ID NO: 13

[0150]

[0151] SEQ ID NO: 19

[0152]

[0153] SEQ ID NO: 24

[0154]

[0155] SEQ ID NO: 27

[0156]

[0157] SEQ ID NO: 35

[0158]

[0159] SEQ ID NO: 39

[0160]

[0161] SEQ ID NO: 42

[0162]

[0163] SEQ ID NO: 45

[0164]

[0165] SEQ ID No: 52

[0166]

[0167] Heavy chain variable domain sequence

[0168] The present disclosure provides an antibody or an antigen-binding fragment thereof, which comprises a heavy chain variable domain that comprises 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.

[0169] SEQ ID NO: 7

[0170]

[0171] SEQ ID NO: 12

[0172]

[0173] SEQ ID NO: 18

[0174]

[0175] SEQ ID NO: 23

[0176]

[0177] SEQ ID NO: 26

[0178]

[0179] SEQ ID NO: 34

[0180]

[0181] SEQ ID NO: 38

[0182]

[0183] SEQ ID NO: 41

[0184]

[0185] SEQ ID NO: 44

[0186]

[0187] SEQ ID NO: 51

[0188]

[0189] The antibodies described herein include fully human antibodies, humanized antibodies, primatized antibodies, and chimeric antibodies. In addition, 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, 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) with the corresponding CDR sequences of the antibodies described herein (e.g., any one of mAb 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).

[0190] In some embodiments, the antibody or antigen-binding fragment is a humanized antibody or antigen-binding fragment that contains one or more or all of the CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, and CDR-L3 sequences described herein, wherein one or more or all of the CDR sequences have at least 70% sequence identity (e.g., 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity) with the corresponding CDR sequences of the humanized antibodies described herein (e.g., any one of mAb 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).

[0191] The antibodies and antibody fragments described herein further 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, wherein one or more or all of the CDR sequences contain one or more (e.g., up to 3) amino acid substitutions (e.g., one or more conservative amino acid substitutions) relative to the corresponding CDR sequences of the antibodies described herein. For example, the antibodies described herein can be generated by incorporating any one or more of the CDR sequences described herein into the framework regions (e.g., FW1, FW2, FW3, and FW4) of a human antibody.

[0192] For example, one strategy that can be used to design the humanized antibodies described herein is to align the sequences of the heavy chain variable region and the light chain variable region of the antibodies described herein with the heavy chain variable region and the light chain variable region of a consensus human antibody. Consensus human heavy and light chain sequences are known in the art (see, e.g., the "VBASE" germline sequence database; see also Kabat, et al., Sequences of Proteins of Immunological Interest, Fifth Edition, 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 which are incorporated herein by reference). In this way, variable domain framework residues and CDRs can be identified by sequence alignment (see Kabat, supra). For example, one or more of the CDRs of a consensus human antibody can be replaced with the corresponding CDRs of the antibodies described herein to generate a humanized antibody. Exemplary variable domains of a consensus human antibody include the heavy chain variable domain, which is identified in U.S. Patent No. 6,054,297; the disclosure of this patent is incorporated herein by reference. These amino acid substitutions can be made, for example, by recombinant expression in a host cell of polynucleotides encoding the heavy and light chains of the humanized antibody using methods known in the art or described herein.

[0193] Similarly, this strategy can also be used to generate primatized antibodies, since one or more or all of the CDRs of, for example, a consensus primate antibody sequence can be replaced with one or more or all of the CDRs of the antibodies described herein. 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 disclosure of each of which is incorporated herein by reference).

[0194] In some embodiments, in addition to the CDR sequences, it may be desirable to introduce specific framework residues from an antibody (e.g., an antibody described herein) into the heavy and / or light chain variable domains of a human antibody. For example, U.S. Patent No. 6,054,297 identifies several instances in which it can be advantageous to retain certain framework residues of a particular antibody heavy or light chain variable region in the resulting humanized antibody. In some embodiments, framework residues can engage an antigen in a non-covalent interaction and thus contribute to the affinity of the antibody for the target antigen. In some embodiments, individual framework residues can modulate the conformation of the CDRs and thus indirectly affect the interaction of the antibody with the antigen. Certain framework residues can form an interface between the VH and VL domains and can thus contribute to the overall antibody structure. In some cases, framework residues can constitute functional glycosylation sites (e.g., Asn-X-Ser / Thr), which can determine antibody structure and antigen affinity when attached to a carbohydrate moiety. In circumstances such as those described above, it can be beneficial to retain certain framework residues of the antibodies described herein in, for example, humanized or primatized antagonistic antibodies or antigen-binding fragments thereof, as the various framework residues can promote high epitope affinity and improvement of the biochemical activity of the antibody or antigen-binding fragment thereof.

[0195] The antibodies described herein also include antibody fragments, Fab domains, F(ab’) molecules, F(ab’)2 molecules, single-chain variable fragments (scFv), tandem scFv fragments, diabodies, triabodies, bispecific variable domain immunoglobulins, multispecific antibodies, bispecific antibodies, and heterospecific antibodies that contain 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, 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) with the corresponding CDR sequences of the antibodies described herein. The antibodies described herein also 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, wherein one or more or all of the CDR sequences contain one or more (e.g., up to 3) amino acid substitutions (e.g., one or more conservative amino acid substitutions) relative to the corresponding CDR sequences of the antibodies described herein. These molecules can be recombinantly expressed, for example, by incorporating polynucleotides encoding these proteins into expression vectors to transfect eukaryotic or prokaryotic cells using techniques described herein or known in the art, or can be chemically synthesized, for example, by solid-phase peptide synthesis methods described herein or known in the art.

[0196] The polypeptides described herein additionally include antibody-like scaffolds that contain, for example, 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, 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) with the corresponding CDR sequences of the antibodies described herein, or contain one or more (e.g., up to 3) amino acid substitutions (e.g., one or more conservative amino acid substitutions) relative to the corresponding CDR sequences of the antibodies described herein. Examples of antibody-like scaffolds include proteins containing the tenth fibronectin type III domain ( 10 Fn3) that contain BC, DE, and FG structural loops similar to those of a typical antibody. 10 The tertiary structure of the Fn3 domain is similar to the tertiary structure of the variable region of the IgG heavy chain, and those skilled in the art can, by 10The residues of the BC, DE, and FG loops of Fn3 are replaced with the residues of the corresponding CDR sequences of the antibodies or antigen-binding fragments thereof described herein, and one or more or all of the CDR sequences of, for example, the antibodies or antigen-binding fragments thereof described herein or sequences having at least 70% sequence identity (e.g., 75%, 80%, 85%, 90%, 95%, 97%, 99%, or 100% sequence identity) with any one or more of these CDR sequences or sequences containing amino acid substitutions (e.g., conservative or non-conservative amino acid substitutions (e.g., up to 3 amino acid substitutions)) relative to any one or more of these CDR sequences are grafted onto the fibronectin scaffold. This can be achieved by recombinant expression of the modified 10 Fn3 domain in prokaryotic or eukaryotic cells (e.g., using the vectors and techniques described herein). The use of 10 the Fn3 domain as an antibody-like scaffold for grafting the CDRs of an antibody onto the BC, DE, and FG structural loops is reported in WO 2000 / 034784, WO 2009 / 142773, WO 2012 / 088006, and U.S. Patent No. 8,278,419; the disclosures of each of which are incorporated herein by reference.

[0197] II. Nucleic Acids and Expression Systems

[0198] 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 recombinant expression of immunoglobulin light and heavy chain genes in a host cell. For recombinant expression of an antibody, the host cell can be transfected with one or more recombinant expression vectors carrying DNA fragments encoding the immunoglobulin light and heavy chains of the antibody such that the light and heavy chains are expressed in the host cell and optionally secreted into the medium in which the host cell is cultured, from which the antibody can be recovered. The antibody heavy and light chain genes are obtained using standard recombinant DNA methods, incorporated into recombinant expression vectors, and the vectors are introduced into host cells, such as those described in Molecular Cloning; A Laboratory Manual, Second Edition (Sambrook, Fritsch and Maniatis (eds), Cold Spring Harbor, N.Y., 1989), Current Protocols in Molecular Biology (Ausubel et al., eds., Greene Publishing Associates, 1989), and U.S. Patent No. 4,816,397; the disclosures of each of which are incorporated herein by reference.

[0199] A vector for expressing an anti-ITGA11 antibody

[0200] Viral genomes provide a rich source of vectors that can be used to efficiently deliver foreign genes into the genomes of cells (e.g., eukaryotic or prokaryotic cells). Viral genomes are particularly useful gene delivery vectors because the polynucleotides contained in such genomes are typically incorporated into the genome of the target cell by generalized or specialized transduction. These processes occur as part of the natural viral replication cycle and do not require the addition of 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 virus), coronaviruses, negative-strand RNA viruses such as orthomyxoviruses (e.g., influenza virus), rhabdoviruses (e.g., rabies and vesicular stomatitis virus), 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 virus, modified vaccinia virus Ankara (MVA), fowlpox virus, and canarypox virus). Other viruses that can be used to deliver polynucleotides encoding the antibody light and heavy chains or antibody fragments described herein include, for example, Norwalk virus, togavirus, flavivirus, reoviruses, papovavirus, hepadnavirus, and hepatitis virus. Examples of retroviruses include: avian leukosis-sarcoma virus, mammalian C-type virus, B-type virus, D-type virus, HTLV-BLV group, lentivirus, spumavirus (Coffin, J.M., Retroviridae: The viruses and their replication, In Fundamental Virology, Third Edition, B.N. Fields, et al., Eds., Lippincott-Raven Publishers, Philadelphia, 1996). Other examples include murine leukemia virus, murine sarcoma virus, mouse mammary tumor virus, bovine leukemia virus, feline leukemia virus, feline sarcoma virus, avian leukemia virus, human T-cell leukemia virus, baboon endogenous virus, gibbon ape leukemia virus, Mason Pfizer monkey virus, simian immunodeficiency virus, simian sarcoma virus, Rous sarcoma virus, and lentivirus. Other examples of vectors are described, for example, in McVey et al., (U.S. Patent No. 5,801,030); the disclosure of each of which is incorporated herein by reference.

[0201] Genome editing technology

[0202] In addition to viral vectors, a variety of other methods have been developed for incorporating genes, such as genes encoding antibody light and heavy chains, single-chain polypeptides, single-chain variable fragments (scFv), tandem scFv, Fab domains, F(ab’)2 domains, diabodies, and triabodies (such as those described herein), into the genome of target cells for polypeptide expression. One such method that can be used to incorporate polynucleotides encoding antibodies or fragments thereof (such as those described herein) into prokaryotic or eukaryotic cells includes transposons. A transposon is a polynucleotide encoding a transposase and containing a polynucleotide sequence or gene of interest flanked by excision sites at the 5’ and 3’ positions. After the transposon is delivered into the cell, the transposase gene begins to express and produces an active enzyme that cleaves the gene of interest from the transposon. This activity is mediated by the site-specific recognition of the transposon excision sites by the transposase. In some embodiments, these excision sites can be terminal repeats or inverted terminal repeats. After excision from the transposon, the gene of interest can be integrated into the genome of prokaryotic or eukaryotic cells through cleavage at similar excision sites present within the nuclear genome catalyzed by the transposase. This results in the insertion of the gene encoding the antibody or its fragment or domain at the excision site in the cleaved nuclear DNA, and subsequent ligation of the phosphodiester bonds to join the gene of interest and the genomic DNA of the prokaryotic or eukaryotic cell, completing the incorporation process. In some embodiments, the transposon can be a retrotransposon such that the gene encoding the antibody is first transcribed into an RNA product and then reverse transcribed into DNA before being incorporated into the genome of prokaryotic or eukaryotic cells. Exemplary transposon systems include the piggybac transposon (described in detail in WO 2010 / 085699) and the sleeping beauty transposon (described in detail in US20050112764); the disclosures of each are incorporated herein by reference.

[0203] 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 clustered regularly interspaced short palindromic repeats (CRISPR) / Cas system, which is a system that originally evolved as an adaptive defense mechanism against viral infection in bacteria and archaea. The CRISPR / Cas system consists of palindromic repeats within plasmid DNA and the associated Cas9 nuclease. This DNA and protein assembly directs site-specific DNA cleavage of a target sequence by first integrating foreign DNA into the CRISPR locus. The polynucleotide containing these foreign sequences and the repeat-spacer elements of the CRISPR locus is then transcribed in the host cell to produce a guide RNA, which can subsequently anneal to the target sequence and localize the Cas9 nuclease to that site. In this way, highly site-specific Cas9-mediated DNA cleavage can be generated in the foreign polynucleotide, because the interaction that brings Cas9 into close proximity to the target DNA molecule is controlled by RNA:DNA hybridization. Thus, in theory, the CRISPR / Cas system can be designed to cleave any target DNA molecule of interest. This technique has been utilized to edit eukaryotic genomes (Hwang et al., Nat. Biotech., 31:227-229, 2013), and can be used as an effective method for site-specifically editing the genomes of eukaryotes or prokaryotes to cleave DNA prior to incorporation of a polynucleotide encoding an antibody or fragment thereof described herein. The use of CRISPR / Cas for regulating gene expression has been described in U.S. Patent No. 8,697,359, the disclosure of which is incorporated herein by reference.

[0204] Alternative methods for site-specifically cleaving genomic DNA prior to incorporation of a polynucleotide encoding an antibody or fragment thereof (such as those described herein) include the use of zinc finger nucleases and transcription activator-like effector nucleases (TALEN). Unlike the CRISPR / Cas system, these enzymes do not contain a guide polynucleotide to localize to a specific target sequence. Target specificity is instead controlled by the DNA-binding domains within these enzymes. Zinc finger nucleases and TALEN for genome editing applications are described in Urnov et al. (Nat. Rev. Genet., 11:636-646, 2010); and Joung et al., (Nat. Rev. Mol. Cell Biol. 14:49-55, 2013); which are incorporated herein by reference. Other genome editing techniques that can be used to incorporate a polynucleotide encoding an antibody described herein into the genome of prokaryotic or eukaryotic cells include the use of ARCUS, which can be rationally designed to site-specifically cleave genomic DNA TMMeganucleases. Given the established structure-activity relationships for such enzymes, it is particularly advantageous to use these enzymes to incorporate polynucleotides encoding the antibodies or fragments thereof described herein into the genome of prokaryotic or eukaryotic cells. Thus, single-stranded meganucleases can be modified at certain amino acid positions to produce nucleases that selectively cleave DNA at desired positions. These single-stranded nucleases have been extensively described in U.S. Patent Nos. 8,021,867 and 8,445,251; the disclosures of each are incorporated herein by reference.

[0205] Polynucleotide sequence element

[0206] To express an antibody or fragment thereof (such as those described herein), polynucleotides encoding the partial or full-length light and heavy chains (e.g., polynucleotides encoding one or more or all of the CDR sequences of the antibodies or antigen-binding fragments thereof described herein) can be inserted into an expression vector such that the gene is operably linked to transcriptional and translational control sequences. The expression vector and expression control sequences are selected to be compatible with the expression host cell used. Using established techniques described herein or known in the art, the polynucleotides encoding the light and heavy chains of the antibody or fragment thereof can be inserted into separate vectors, or optionally, the two polynucleotides can be incorporated into the same expression vector.

[0207] In addition to the polynucleotides encoding the antibody heavy and light chains (or polynucleotides encoding single-chain polypeptides, antibody fragments (such as scFv molecules) or constructs described herein), the recombinant expression vectors described herein can also carry regulatory sequences that control the expression of the antibody chain genes in the 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 level of protein expression. For example, regulatory sequences suitable for mammalian host cell expression include viral elements that direct high-level protein expression in mammalian cells, such as promoters and / or enhancers derived from cytomegalovirus (CMV) (e.g., CMV promoter / enhancer), simian virus 40 (SV40) (e.g., SV40 promoter / enhancer), adenovirus (e.g., adenovirus major late promoter (AdMLP)), and polyomavirus. The viral regulatory elements and their sequences 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 are incorporated herein by reference.

[0208] In addition to the antibody chain genes and regulatory sequences, the recombinant expression vectors described herein may also carry additional sequences, such as sequences that regulate the replication of the vector in a host cell (e.g., an origin of replication) and selectable marker genes. Selectable marker genes facilitate the selection of host cells into which the vector has been introduced (see, e.g., U.S. Patent Nos. 4,399,216, 4,634,665, and 5,179,017). For example, selectable marker genes typically confer resistance to cytotoxic drugs (e.g., G418, puromycin, blasticidin, hygromycin, or methotrexate) on host cells into which the vector has been introduced. Suitable selectable marker genes include the dihydrofolate reductase (DHFR) gene (for methotrexate selection / amplification in DHFR− host cells) and the neo gene (for G418 selection). To express the light and heavy chains of an antibody or fragment thereof, expression vectors containing polynucleotides encoding the heavy and light chains can be transfected into host cells by standard techniques.

[0209] Host cells for expressing an anti-ITGA11 antibody or fragment thereof

[0210] It is possible to express the antibodies or fragments thereof described herein in prokaryotic or eukaryotic host cells. In certain embodiments, the expression of the antibody or fragment thereof is carried out in eukaryotic cells (e.g., mammalian host cells) for proper folding and optimal secretion of immunologically active antibodies. Exemplary mammalian host cells for expressing the recombinant antibodies or antigen-binding fragments described herein include Chinese hamster ovary cells (CHO cells) (including DHFR− CHO cells, which are described in Urlaub and Chasin (1980, Proc. Natl. Acad. Sci. USA 77:4216-4220) and are used with a DHFR selectable marker, e.g., 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 can be used to express antibodies and fragments thereof include bacterial cells, such as BL-21(DE3) Escherichia coli (E. coli) cells, which can be transformed with a vector containing foreign DNA according to established protocols. Additional eukaryotic cells that can be used to express antibodies include yeast cells, such as auxotrophic strains of Saccharomyces cerevisiae, which can be transformed according to established procedures known in the art and selectively grown in minimal 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 period of time sufficient to allow expression of the antibody in the host cell or secretion of the antibody into the medium in which the host cell is growing.

[0211] Antibodies or antigen-binding fragments thereof can be recovered from culture media using standard protein purification methods. Host cells can also be used to produce portions of a full-length antibody, such as Fab fragments or scFv molecules. Also included herein are methods of altering the above procedures according to established protocols known in the art. For example, it may be desirable to transfect a host cell with DNA encoding the light or heavy chain (but not both) of an antibody or fragment thereof described herein to produce an antigen-binding fragment of the antibody.

[0212] After production by recombinant expression, the antibodies or fragments thereof described herein can be purified by any method known in the art, such as methods useful for purifying immunoglobulin molecules, e.g., by chromatography (e.g., ion exchange chromatography, affinity chromatography, and size exclusion chromatography), centrifugation, differential solubility, or by any other standard technique for purifying proteins. In addition, the antibodies or fragments thereof described herein can be fused with heterologous polypeptide sequences described herein or known in the art to facilitate purification or to generate therapeutic conjugates.

[0213] Following isolation, if desired, the antibody or fragment thereof can be further purified, e.g., by high performance liquid chromatography (see, e.g., Fisher, Laboratory Techniques in Biochemistry and Molecular Biology (Work and Burdon, eds., Elsevier, 1980; which is incorporated herein by reference)); or by gel filtration chromatography, e.g., on a SUPERDEX TM 75 column (Pharmacia Biotech AB, Uppsala, Sweden).

[0214] III. Generation and Purification of Anti-ITGA11 Antibodies or Fragments Thereof

[0215] Antigens that contain epitopes corresponding to portions of ITGA11 can be used to generate, for example, monoclonal antibodies, polyclonal antibodies, chimeric antibodies, humanized antibodies, or recombinant ITGA11-specific antibodies. Methods include immunological methods 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,” in Burdon et al., Eds., Laboratory Techniques in Biochemistry and Molecular Biology, Volume 13, Elsevier Science Publishers, Amsterdam, 1985), and recombinant DNA methods described by Huse et al. (Science 246:1275-1281, 1989).

[0216] Briefly, the antigen can be administered to a host animal (e.g., rabbit, mouse, rat, goat, guinea pig, hamster, horse, and sheep, as well as non-human primates) in combination with an adjuvant. Administration of such antigens can be accomplished by any of a variety of methods, including but not limited to subcutaneous or intramuscular injection. After administration, the results of the antibody titers generated in the host animal are monitored, which can be done by any of a variety of techniques well known in the art (e.g., conventional bleeding), where antiserum is isolated (e.g., by centrifugation) and then screened to determine the presence of antibodies having binding affinity for, for example, ITGA11. Screening for the desired antibodies can be done by techniques that include, for example, radioimmunoassay, ELISA, sandwich immunoassay, immunoradiometric assay, gel diffusion precipitation reaction, in situ immunoassay (e.g., using colloidal gold, enzyme, or radioisotope labeling), 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.

[0217] The resulting antiserum from the host animal can be affinity purified to obtain the antibodies of the present disclosure. The antiserum can be purified by conventional techniques, such as introducing the antiserum into a separation column. The antigen of the present disclosure can be immobilized on the column to separate and purify the antibodies. Then, the column can be washed to remove antibodies that are not specific to the antigen immobilized on the column, and the remaining antibodies are finally eluted from the column. The separated antibodies can then be stored by conventional methods known to those skilled in the art.

[0218] Established procedures for immunizing primates are known in the art (see, for example, WO 1986 / 6004782; which is incorporated herein by reference). Immunization represents a robust method for generating monoclonal antibodies by exploiting the antigen specificity of B lymphocytes. For example, monoclonal antibodies can be prepared by the Kohler-Millstein procedure (which is described, for example, in EP 0110716; which is incorporated herein by reference), in which spleen cells from a non-human animal (e.g., a primate) are administered a peptide with the antigenic peptide. Clonally expanded B lymphocytes generated by immunization can be isolated from the animal serum and subsequently fused with myeloma cells to form hybridomas. Hybridomas are particularly useful vehicles 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 (e.g., purifying the antibodies from the cell culture medium by affinity chromatography).

[0219] Optionally, an antibody library (e.g., a naive antibody library, a synthetic antibody library, a semi-synthetic antibody library, or a combinatorial library) can be screened to identify antibodies. Such libraries are commercially available from a number of sources (e.g., Cambridge Antibody, Cambridge, United Kingdom, Genetastix Corporation, Pacific Northwest Laboratory, Richland, Washington, 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, which are incorporated herein by reference.

[0220] The screening of an antibody library can be carried out by using one of the methods known to those skilled in the art, said methods including, for example, phage display, selective infection phage, polysome technology, and assay systems for enzyme activity or protein stability. Antibodies with desired properties can be identified, for example, by sequencing of the corresponding nucleic acid sequence, amino acid sequencing, or mass spectrometry. Optimization is carried out by replacing subsequences with different sequences (e.g., random sequences), and then repeating the screening step one or more times. Antibodies can be screened for, for example, optimized affinity or specificity for a target molecule, optimized expression yield, optimized stability, or optimized solubility.

[0221] 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 M or greater.

[0222] IV. Anti-ITGA11 Antibody Conjugates

[0223] It may be desirable to conjugate an antibody or a fragment thereof with a second molecule, for example, to modulate the activity of the antibody in vivo or for diagnostic purposes. Any of a variety of established conjugation strategies well known in the art can be used to conjugate an antibody or its antigen-binding fragment to other molecules at the N-terminus or C-terminus of the antibody light chain or heavy chain. Examples of pairs of reactive functional groups that can be used to covalently tether an antibody or its antigen-binding fragment to another molecule include, but are not limited to, thiol pairs, carboxylic acid and amino, ketone and amino, aldehyde and amino, thiol and α,β-unsaturated moieties (e.g., maleimide or dehydroalanine), thiol and α-haloamide, carboxylic acid and hydrazide, aldehyde and hydrazide, and ketone and hydrazide.

[0224] An antibody or antigen-binding fragment thereof can be directly covalently attached to another molecule by chemical conjugation as described. Optionally, a fusion protein containing the antibody or antigen-binding fragment thereof can be recombinantly expressed from a cell (e.g., a eukaryotic or prokaryotic cell). 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. The linker can then be conjugated to another molecule, or the linker can be conjugated to another molecule prior to its attachment to the antibody or antigen-binding fragment thereof. Examples of linkers that can be used to form conjugates include polypeptide linkers, such as linkers containing naturally occurring or non-naturally occurring amino acids. In some embodiments, it may be desirable to include D-amino acids in the linker, as these residues are not present in naturally occurring proteins and are thus more resistant to degradation by endogenous proteases. Fusion proteins containing polypeptide linkers can be prepared using chemical synthesis techniques (e.g., techniques described herein) or by recombinantly expressing 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 well known in the art and can be cleaved by enzymatic hydrolysis, photolysis, hydrolysis under acidic conditions, hydrolysis under alkaline conditions, oxidation, disulfide reduction, nucleophilic cleavage, or organometallic cleavage, depending on the reactive components of the linker (Leriche et al., Bioorg. Med. Chem., 20:571-582, 2012).

[0225] The antibodies or antigen-binding fragments thereof described herein can be conjugated with, mixed with, or administered separately from a therapeutic agent.

[0226] Labeled anti-ITGA11 antibody or antigen-binding fragment thereof

[0227] In some embodiments, the antibodies or antigen-binding fragments thereof described herein are conjugated to another molecule (e.g., an epitope tag) for purposes of purification or detection. Examples of such molecules useful for protein purification include those that present a structural epitope that can be recognized by a second molecule. This is a common strategy employed in protein purification by affinity chromatography, in which the molecule is immobilized on a solid support and exposed to a heterogeneous mixture containing a target protein conjugated to a molecule that can bind the immobilized compound. Examples of epitope tag molecules that can be conjugated to the antibodies or antigen-binding fragments thereof described herein for purposes of molecular recognition include, but are not limited to, maltose-binding protein, glutathione-S-transferase, polyhistidine tag, FLAG tag, myc tag, human influenza hemagglutinin (HA) tag, biotin, streptavidin. Conjugates 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, the antibodies or fragments thereof described herein conjugated to an epitope tag can be purified from a complex mixture of other proteins and biomolecules (e.g., DNA, RNA, carbohydrates, phospholipids, etc.) by treating the mixture with a solid-phase resin that contains a complementary molecule that can selectively recognize and bind the epitope tag of the antibody or fragment. Examples of solid-phase resins include agarose beads, which are compatible with purification in an aqueous solution.

[0228] The antibodies or antigen-binding fragments thereof described herein can also be covalently attached to a fluorescent molecule, e.g., to detect the antibody or antigen-binding fragment by fluorescence assays and / or by direct visualization using fluorescence microscopy. Exemplary 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, hoescht, 4’,6-diamidino-2-phenylindole (DAPI), propidium iodide, fluorescein, coumarin, rhodamine, tetramethylrhodamine, and cyanine. Additional examples of fluorescent molecules suitable for conjugation to the antibodies described herein are well known in the art and have been described in detail, for example, in U.S. Patent Nos. 7,417,131 and 7,413,874, each of which is incorporated herein by reference.

[0229] Antibodies or antigen-binding fragments thereof that contain a fluorescent molecule are particularly useful for monitoring the cell surface localization properties of the antibodies and their fragments described herein. For example, cultured mammalian cells can be exposed to an antibody or antigen-binding fragment thereof described herein that has been covalently conjugated to a fluorescent molecule, and these cells can then be 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 a tagged antibody because individual planes of the cell can be analyzed to distinguish antibodies or their fragments that have been internalized into the cell interior (e.g., by receptor-mediated endocytosis) from antibodies or their fragments that are bound to the outer surface of the cell membrane. Additionally, cells can be treated with an antibody that is conjugated to a fluorescent molecule that emits visible light at a specific wavelength (e.g., fluorescein, which fluoresces at approximately 535 nm) and an additional fluorescent molecule that is known to localize to a specific location on the cell surface and emits fluorescence at a different wavelength (e.g., a molecule that localizes to CD25 and fluoresces at approximately 599 nm). The resulting emission pattern can be visualized by confocal fluorescence microscopy, and images at these two wavelengths can be merged to reveal information about the localization of the antibody or its antigen-binding fragment relative to other receptors on the cell surface.

[0230] Bioluminescent proteins can also be incorporated into fusion proteins for the purpose of detecting and visualizing antibodies or their fragments. Bioluminescent proteins (e.g., 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 of using them are described, for example, in U.S. Patent Nos. 5,292,658, 5,670,356, 6,171,809, and 7,183,092, each of which is incorporated herein by reference. Antibodies or antigen-binding fragments thereof labeled with a bioluminescent protein are useful tools for detecting the antibodies described herein after an in vitro assay. 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 the antibody in the mixture of other proteins can be achieved by treating the membrane with an appropriate luciferase substrate and then visualizing the protein mixture on film using established protocols.

[0231] The antibodies or antigen-binding fragments thereof described herein can also be conjugated to a molecule that includes a radioisotope such that the antibodies or fragments described herein can be detected by analyzing the radioactive emission pattern of the isotope. Optionally, the antibody or fragment can be directly modified by incorporating the radioisotope into the antibody during the protein preparation process. Methionine ( 35 S), nitrogen (15 N) or carbon ( 13 C) radioactive isotopes can be incorporated into the antibodies or fragments thereof described herein by culturing bacteria, for example, in a medium supplemented with nutrients containing these isotopes. Optionally, tyrosine derivatives containing radioactive halogens can be incorporated into the antibody by culturing bacterial cells, for example, in a medium supplemented with radiolabeled tyrosine. Tyrosine functionalized with a radioactive halogen at the C2 position of the phenol system has been shown to be rapidly incorporated into extended polypeptide chains in vivo using endogenous translation enzymes (U.S. Patent No. 4,925,651; which is incorporated herein by reference). Halogens include fluorine, chlorine, bromine, iodine, and astatine. In addition, the antibody can be modified by functionalizing the polypeptides described herein with a radioactive isotope after isolation and purification from the cell culture. Halogens represent a class of isotopes that can be readily incorporated into purified proteins by aromatic substitution of tyrosine or tryptophan (e.g., by reaction of one or more of these residues with an 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.

[0232] Another alternative strategy for incorporating radioactive isotopes is to covalently attach a chelating group to the antibody or fragment or construct thereof. The chelating group can be covalently attached to the antibody or fragment by attachment to a reactive functional group (e.g., thiol, amino, alcohol, or carboxylic acid). The chelating group can then be modified to contain any of a variety of metal radioactive isotopes, including but not limited to such as 125 I, 67 Ga, 111 In, 99 Tc, 169 Yb, 186 Re, 123 I, 124 I, 125 I, 131 I, 99m Tc, 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 radionuclides of Ac.

[0233] In some embodiments, it may be desirable to covalently conjugate an antibody or fragment thereof described herein with a chelating group capable of binding a heavy element or a rare earth ion (such as Gd 3+ , Fe 3+ , Mn 3+ or Cr 2+ ). Conjugates containing chelating groups coordinated to such paramagnetic metals are very useful in MRI imaging applications. Paramagnetic metals include, but are not limited to, 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 that binds 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 the patient by any of the administration routes described herein (e.g., intravenous administration), and then analyzing the location of the administered antibody by recording the patient's MRI according to an established protocol. The antibody or its antigen-binding fragment can additionally be conjugated to other molecules for the purpose of increasing the solubility and stability of the protein in aqueous solutions. Examples of such molecules include PEG, PSA, bovine serum albumin (BSA), and human serum albumin (HSA), etc. For example, an antibody can be conjugated to a carbohydrate moiety to enable the antibody or its fragment to escape detection by the immune system of a patient undergoing treatment. This hyperglycosylation process reduces the immunogenicity of the therapeutic protein by sterically inhibiting the interaction of the protein with B cell receptors in the circulation. Optionally, the antibody or its fragment can be conjugated to a molecule that prevents clearance from human serum and improves the pharmacokinetic characteristics of the antibodies described herein. Exemplary molecules that can be conjugated to or inserted into the antibodies or antigen-binding fragments described herein to attenuate clearance and improve the pharmacokinetic characteristics of these antibodies and fragments include salvage receptor binding epitopes. These epitopes are present in the Fc region of IgG immunoglobulins and have been shown to bind to Fc receptors and extend the half-life of antibodies in human serum. Insertion of salvage receptor binding epitopes into antibodies or their fragments can be achieved as described in U.S. Patent No. 5,739,277; this patent is incorporated herein by reference.

[0234] V. Methods of Treatment

[0235] 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 fibrotic disorders, inflammatory disorders, or cancer. The anti-ITGA11 antibodies or antigen-binding fragments thereof can be administered to a mammalian subject, such as a human, suffering from a fibrotic disorder, an inflammatory disorder, or cancer.

[0236] Fibrotic disorders

[0237] Fibrosis is a common response to a range of tissue injuries and can lead to organ dysfunction. Diseases characterized by pathological fibrosis and treatable using the methods and compositions of the present invention include, but are not limited to, liver fibrosis (e.g., fibrosis associated with cirrhosis (e.g., alcohol-induced cirrhosis, virus-induced cirrhosis, post-hepatitis C cirrhosis, and primary biliary cirrhosis), schistosomiasis, cholangitis (e.g., sclerosing cholangitis), and autoimmune-induced hepatitis), renal fibrosis (e.g., tubulointerstitial fibrosis, scleroderma, diabetic nephropathy, and glomerulonephritis), skin fibrosis (e.g., scleroderma, hypertrophic scars, and keloids, nephrogenic fibrosing dermopathy, and burns), myelofibrosis, neurofibromatosis, fibroma, intestinal fibrosis, and surgical-induced fibrotic adhesions), cardiac fibrosis (e.g., fibrosis associated with myocardial infarction), vascular fibrosis (e.g., fibrosis associated with arterial restenosis and atherosclerosis after angioplasty), ocular fibrosis (e.g., fibrosis associated with after cataract surgery, proliferative vitreoretinopathy, and retro-orbital fibrosis), fibrosis of the bone marrow (e.g., idiopathic myelofibrosis and drug-induced myelofibrosis), pulmonary fibrosis (e.g., pulmonary interstitial fibrosis), glomerulonephritis, heart failure (ischemic and non-ischemic), scleroderma, excessive scar tissue after surgery or device insertion, trauma or burns, progressive kidney disease, heart valve disease, hypertensive heart disease, joint and perijoint fibrosis, myelofibrosis, ocular / vitreous fibrosis, intestinal fibrosis and stricture, peritoneal and retroperitoneal fibrosis, pancreatic fibrosis, nephrogenic systemic fibrosis, primary sclerosing cholangitis, and the development of pathological matrix also plays a role in fibroproliferative tumor progression and metastasis.

[0238] Inflammatory disorders

[0239] Inflammation can be classified as acute or chronic. Acute inflammation is the body's initial response to a harmful stimulus and is achieved by increasing the movement of plasma and white blood cells (especially granulocytes) from the blood into the damaged tissue. A series of biochemical events propagate and mature the inflammatory response, involving the local vascular system, the immune system, and various cells within the damaged tissue. Prolonged inflammation, known as chronic inflammation, results in a progressive shift in the types of cells present at the site of inflammation and is characterized by the simultaneous occurrence of tissue destruction and repair during the inflammatory process.Diseases characterized by pathological inflammation and treatable using the methods and compositions of the present invention include, but are not limited to: asthma (e.g., allergic asthma, exercise-induced asthma, aspirin-sensitive / aggravated asthma, atopic asthma, severe asthma, mild asthma, moderate-to-severe asthma, asthma not previously treated with corticosteroids, chronic asthma, corticosteroid-resistant asthma, corticosteroid-refractory asthma, newly diagnosed and untreated asthma, asthma caused by smoking, asthma uncontrolled by corticosteroids, etc.), airway inflammation, airway hyperresponsiveness, airway hyperreactivity, sinusitis, sinusitis with polyps, nasal polyposis, arthritis (e.g., osteoarthritis, rheumatoid arthritis, collagen-induced arthritis, arthritis due to injury, etc.), eosinophilic inflammation, mast cell-mediated inflammatory diseases, sepsis, septic shock, seronegative enthesopathy and arthropathy (SEA) syndrome, osteoporosis, eosinophilic esophagitis, scleroderma, dermatitis, atopic dermatitis, allergic rhinitis, bullous pemphigoid, urticaria (e.g., chronic urticaria), cartilage inflammation, polymyalgia rheumatica, polyarteritis nodosa, Wegener's granulomatosis, Behcet's disease, myositis, polymyositis, dermatomyositis, vasculitis, arteritis, diabetic nephropathy, interstitial cystitis, graft-versus-host disease (GVHD), gastrointestinal inflammatory conditions (e.g., inflammatory bowel disease (IBD), ulcerative colitis (UC), Crohn's disease (CD), colitis (e.g., colitis caused by environmental damage (e.g., caused by or associated with treatment regimens (e.g., chemotherapy, radiotherapy, etc.)), infectious colitis, ischemic colitis, collagenous or lymphocytic colitis, necrotizing enterocolitis, colitis in conditions such as chronic granulomatous disease or celiac disease, food allergy, gastritis, infectious gastritis or enterocolitis (e.g., chronic active gastritis infected with Helicobacter pylori) and other forms of gastrointestinal inflammation caused by infectious pathogens), and inflammatory lung conditions (e.g., chronic obstructive pulmonary disease (COPD), eosinophilic lung inflammation, infection-induced lung conditions (including those associated with viral (e.g., influenza, parainfluenza virus, rotavirus, human metapneumovirus and respiratory syncytial virus), bacterial, fungal (e.g., Aspergillus), parasitic or prion infections), allergen-induced lung conditions, pollutant-induced lung conditions (e.g., asbestosis, silicosis or berylliosis), gastric aspiration-induced lung conditions, immune dysregulation, inflammatory conditions with a genetic predisposition (e.g., cystic fibrosis), body trauma-induced lung 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)).

[0240] Cancer

[0241] Cancer is a condition characterized by uncontrolled cell growth of a cell population. ITGA11 signaling can contribute to cancer development in humans, including invasion and metastasis. Cancers that can be treated using the methods and compositions of the present invention include, but are not limited to, breast cancer, colorectal cancer, liver cancer, kidney cancer, hepatocellular carcinoma, lung cancer, pancreatic cancer, gastrointestinal cancer, melanoma, ovarian cancer, prostate cancer, cervical cancer, bladder cancer, glioblastoma, head and neck cancer, and cholangiocarcinoma. In some embodiments, the method further comprises administering to the subject at least one additional anti-cancer agent.

[0242] VI. Pharmaceutical Compositions

[0243] 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 containing one or more or all of the CDR sequences of the antibodies or antigen-binding fragments described herein, such as human, humanized, or chimeric variants of anti-ITGA11 described herein.

[0244] The pharmaceutical compositions described herein can contain a combination of the antibodies or antigen-binding fragments thereof described herein and 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 (Remington's Pharmaceutical Sciences 16th edition, Osol, A. Ed. (1980); which is incorporated herein by reference) and in a desired form, such as in the form of a lyophilized preparation or an aqueous solution. The compositions can also be prepared to contain a desired concentration of the active agent. For example, by weight (w / w), the pharmaceutical compositions described herein can 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.

[0245] In addition, the active agent that can be incorporated into a pharmaceutical formulation can itself have a desired level of purity. For example, the antibodies or antigen-binding fragments thereof described herein can be characterized by a particular purity after separation of the antibody from cell culture medium or after chemical synthesis, such as by established solid-phase peptide synthesis methods or native chemical ligation as described herein, of, for example, single-chain antibody fragments (e.g., scFv). The antibodies or antigen-binding fragments thereof described herein can 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) prior to incorporation of the antibody into a pharmaceutical composition.

[0246] The pharmaceutical compositions of the antibodies or antigen-binding fragments thereof described herein can be prepared as lyophilized formulations or aqueous solutions for storage by mixing the antibody having the desired purity with optional pharmaceutically acceptable carriers, excipients or stabilizers (e.g., buffers, stabilizers, preservatives, isotonic agents, non-ionic detergents, antioxidants and various other additives) commonly employed in the art. See, e.g., Remington's Pharmaceutical Sciences, 16th edition (Osol, ed. 1980; which is incorporated herein by reference). Such additives must be non-toxic to the recipient at the doses and concentrations employed.

[0247] Buffer

[0248] Buffers help maintain the pH within a range close to physiological conditions. They can be present at concentrations in the range of about 2 mM to about 50 mM. Suitable buffers for use with the antibodies or antigen-binding fragments thereof described herein include organic and inorganic acids and their salts, such as citrate buffers (e.g., sodium citrate - disodium citrate mixtures, citric acid - trisodium citrate mixtures, citric acid - sodium citrate mixtures, etc.), succinate buffers (e.g., succinic acid - sodium 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 - sodium fumarate mixtures, fumaric acid - disodium fumarate mixtures, sodium 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.). In addition, phosphate buffers, histidine buffers and trimethylamine salts (e.g., Tris) can be used.

[0249] Preservative

[0250] Preservatives can be added to the compositions described herein to retard the growth of microorganisms, and the preservatives can be added in an amount in the range of 0.2% - 1% (w / v). Suitable preservatives for use with the antibodies or antigen-binding fragments thereof described herein include phenol, benzyl alcohol, m-cresol, methylparaben, propylparaben, cetrimonium chloride, benzalkonium halides {e.g., chloride, bromide, and iodide), hexamethonium chloride, and alkyl parabens (e.g., methyl or propyl paraben), catechol, resorcinol, cyclohexanol, and 3-pentanol. Osmotic agents (sometimes referred to as "stabilizers") can be added to ensure the isotonicity of the liquid compositions described herein, and the osmotic agents include polyhydric alcohols, such as trihydric alcohols or higher polyhydric alcohols, such as glycerol, arabitol, xylitol, sorbitol, and mannitol. Stabilizers refer to a broad class of excipients whose functions can range from fillers to additives that solubilize therapeutic agents or help prevent denaturation or adhesion to the container walls. Typical stabilizers can be polyhydric alcohols (listed above); amino acids, such as arginine, lysine, glycine, glutamine, asparagine, histidine, alanine, ornithine, L-leucine, 2-phenylalanine, glutamic acid, threonine, etc., organic sugars or sugar alcohols, such as lactose, trehalose, stachyose, mannitol, sorbitol, xylitol, ribitol, myoinisitol, galactitol, glycerol, etc., including cycloalcohols (e.g., inositol); polyethylene glycol; amino acid polymers; sulfur-containing reducing agents, such as urea, glutathione, lipoic acid, sodium thioglycolate, thioglycerol, a-monothioglycerol, and sodium thiosulfate; low molecular weight polypeptides (e.g., peptides of 10 residues or fewer); proteins, such as human serum albumin, bovine serum albumin, gelatin, or immunoglobulins; hydrophilic polymers, such as polyvinylpyrrolidone monosaccharides, such as xylose, mannose, fructose, glucose; disaccharides (e.g., lactose, maltose, sucrose) and trisaccharides (e.g., raffinose); and polysaccharides (e.g., dextran). The stabilizer can be present in a range of 0.1 to 10,000 weights per weight of the active protein per portion.

[0251] Detergent

[0252] Nonionic surfactants or detergents (also referred to as "wetting agents") can be added to aid in solubilizing the therapeutic agent, as well as to protect the therapeutic protein from agitation-induced aggregation, which also allows the formulation to be exposed to shear surface stress without causing protein denaturation. Suitable nonionic surfactants include polysorbates (20, 80, etc.), poloxamers (184, 188, etc.), Pluronic polyols, polyoxyethylene sorbitan monoethers ( etc.). The non-ionic surfactant may be present in the range of about 0.05 mg / mL to about 1.0 mg / mL (such as about 0.07 mg / mL to about 0.2 mg / mL).

[0253] Various other excipients include fillers (such as starch), chelating agents (such as EDTA), antioxidants (such as ascorbic acid, methionine, vitamin E), and co-solvents.

[0254] Other pharmaceutical carriers

[0255] Optional pharmaceutically acceptable carriers that can be incorporated into the pharmaceutical compositions described herein may include dextrose, sucrose, sorbitol, mannitol, starch, arable rubber, potassium phosphate, arginine salts, gelatin, potassium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methylcellulose, methylparaben, propylparaben, talc, magnesium stearate, and mineral oil, but are not limited thereto. Compositions containing the antibodies described herein may further comprise lubricants, humectants, sweeteners, flavoring agents, emulsifying agents, suspending agents, and preservatives. Details of suitable pharmaceutically acceptable carriers and formulations can be found in Remington's Pharmaceutical Sciences (19th ed., 1995), which is incorporated herein by reference.

[0256] VII. Routes of administration and dosage

[0257] The anti-ITGA11 antibodies or antigen-binding fragments thereof described herein can be administered to mammalian subjects (such as humans) by various routes (such as oral, transdermal, subcutaneous, intranasal, intravenous, intramuscular, intraocular, intratumoral, parenteral, topical, intrathecal, and intraventricular) to treat, for example, the diseases and conditions described herein (such as fibrotic disorders, inflammatory disorders, or cancer). The most suitable route of administration in any given case will depend on the specific polypeptide being administered, the patient, the method of drug formulation, the method of administration (such as the time and route of administration), the age, weight, and sex of the patient, the severity of the disease being treated, the patient's diet, and the patient's excretion rate.

[0258] A physician having ordinary skill in the art can readily determine an effective amount of an anti-ITGA11 antibody or an antigen-binding fragment thereof for administration to a mammalian subject in need thereof (e.g., a human). For example, the physician can begin 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, the physician can begin a treatment regimen by administering the antibody or an antigen-binding fragment thereof described herein at a high dose and then administering progressively lower doses until the therapeutic effect is achieved. Generally, the appropriate daily dose of the antibody or an antigen-binding fragment thereof will be the amount of the compound that is the lowest dose effective to produce a therapeutic effect. The antigen-binding fragments described herein can be administered, for example, by injection (e.g., by intravenous, intramuscular, intraperitoneal, or subcutaneous injection), optionally proximal to the site of the target tissue. The daily dose of the therapeutic composition of the antibody described herein can be administered as a single dose, or as two, three, four, five, six, or more doses administered at appropriate intervals over one day, one week, one month, or one year, or optionally in unit dose form as needed. The antibody described herein can be administered alone, or it can be administered in combination with excipients, carriers, and optionally additional therapeutic agents as a pharmaceutical formulation.

[0259] The effective dose range of the anti-ITGA11 antibody or an antigen-binding fragment thereof described herein can be, for example, from about 0.0001 to about 100 mg / kg body weight per single (e.g., bolus) administration, multiple administrations, or continuous administration (e.g., continuous infusion), or to a serum concentration of 0.0001 - 5000 μg / mL serum concentration per single (e.g., bolus) administration, multiple administrations, or continuous administration (e.g., continuous infusion), or any effective range or value therein, depending on the condition being treated, the route of administration, and the age, weight, and condition of the subject. In certain embodiments, the range for each dose can be from about 0.0001 mg to about 500 mg / kg body weight. For example, the pharmaceutical composition described herein can be administered at a daily dose in the range of 0.001 - 100 mg / kg (body weight). The dose can be administered once or more (e.g., 2 - 10 times) per day, week, month, or year to a mammalian subject in need thereof (e.g., a human).

[0260] The anti-ITGA11 antibody or antigen-binding fragment thereof can be administered to a patient by continuous intravenous infusion or by bolus injection. The antibody or antigen-binding fragment thereof can be administered to the patient in an amount of, for example, 0.01 μg to about 5 g in a volume of 10 μL to 10 mL. The antibody or antigen-binding fragment thereof can be administered to the patient over a time range of from a few minutes to a few hours. For example, the antibody or antigen-binding fragment thereof described herein can be administered over a time range of 5 minutes to 5 hours, such as at 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, 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.

[0261] When the anti-ITGA11 antibody or antigen-binding fragment thereof is administered to a patient in combination with another therapeutic agent, the antibody or antigen-binding fragment thereof and the other therapeutic agent can be co-administered to the patient, for example, by continuous intravenous infusion or bolus injection of the first agent followed by continuous intravenous infusion or bolus injection of the second agent. The administration of the two agents can be carried out simultaneously. Optionally, the administration of the antibody or antigen-binding fragment thereof can be carried out before or after the administration of the other therapeutic agent. In some embodiments, the administration of the second agent (e.g., the antibody or antigen-binding fragment thereof) begins about 5 minutes to about 4 weeks or longer after the end of the administration of the first agent (e.g., the other therapeutic agent). For example, the administration of the second agent can begin 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 longer after the end of the administration of the first agent.

[0262] Therapeutic compositions can be administered using medical devices known in the art. For example, in embodiments, the therapeutic compositions described herein can be administered using needleless subcutaneous 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 implants and modules that can be used in conjunction with the compositions and methods described herein include: U.S. Patent No. 4,487,603, which discloses an implantable microinfusion pump for dispensing drugs at a controlled rate; U.S. Patent No. 4,486,194, which discloses a therapeutic device for administering drugs through the skin; U.S. Patent No. 4,447,233, which discloses a drug infusion pump for delivering drugs at a precise infusion rate; U.S. Patent No. 4,447,224, which discloses a variable flow rate implantable infusion device for continuous drug delivery; U.S. Patent No. 4,439,196, which discloses an osmotic drug delivery system having multiple compartments; and U.S. Patent No. 4,475,196, which discloses an osmotic drug delivery system. These patents are incorporated herein by reference. Many other such implants, delivery systems, and modules are known to those of skill in the art.

[0263] VIII. Kits Containing Anti-ITGA11 Antibodies or Antigen-Binding Fragments Thereof

[0264] Also included herein are kits containing anti-ITGA11 antibodies or antigen-binding fragments thereof. The kits provided herein can contain any of the antibodies or antigen-binding fragments thereof described above, as well as any polynucleotide encoding these polypeptides, any vector containing these polynucleotides, or any cell (e.g., a prokaryotic or eukaryotic cell) engineered to express and secrete the antibodies described herein.

[0265] The kits described herein can include reagents that can be used to generate the compositions described herein (e.g., anti-ITGA11 antibodies or antigen-binding fragments thereof). Optionally, the kits described herein can include reagents that can induce the expression of an antibody or antigen-binding fragment thereof in a cell (e.g., a mammalian cell), such as doxycycline or tetracycline. In other cases, the kits described herein can contain compounds capable of binding and detecting a fusion protein containing an antibody or antigen-binding fragment thereof and an epitope tag. For example, in such cases, the kits described herein can contain maltose, glutathione, nickel-containing complexes, anti-FLAG antibodies, anti-myc antibodies, anti-HA antibodies, biotin, or streptavidin.

[0266] 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 contain secondary antibodies that recognize the Fc region of the antibody or antigen-binding fragment and are 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, using established immunofluorescence techniques in specific tissues or cultured mammalian cells. In some embodiments, the kits described herein may include additional fluorescent compounds that exhibit known subcellular localization patterns. These reagents can be used in combination with another antibody-fluorophore conjugate (e.g., a conjugate that specifically recognizes a different receptor on the cell surface) to analyze the localization of the antibody or antigen-binding fragment relative to other cell surface proteins.

[0267] The kits described herein may also contain reagents that can be used to analyze a patient's response to treatment with the antibodies or antigen-binding fragments described herein. For example, the kits described herein may include an antibody or antigen-binding fragment and one or more reagents that can be used to determine the amount of T-regulatory cells in a blood sample collected from a subject (e.g., a human) undergoing treatment with the antibodies described herein. The kit may contain, for example, antibodies that selectively bind to cell surface antigens presented by T-regulatory cells (e.g., CD4 and CD25). Optionally, these antibodies may be labeled with a fluorescent dye (e.g., fluorescein or tetramethylrhodamine) to facilitate analysis of T-regulatory cells by fluorescence-activated cell sorting (FACS) methods known in the art. The kits described herein may optionally contain 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 in restoring the proliferation of tumor-infiltrating lymphocytes. For example, the kits described herein may contain antibodies that selectively bind to cell surface markers (e.g., CD8 or CD3) on the surface of cytotoxic T cells. Optionally, these antibodies may be labeled with a fluorescent molecule to enable quantification by FACS analysis.

[0268] The kits described herein may also contain one or more reagents that can be used to determine the affinity and selectivity of the antibodies or antigen-binding fragments described herein for one or more peptides derived from ITGA11. For example, the kit may contain an anti-ITGA11 antibody or antigen-binding fragment and one or more reagents that can be used in an ELISA assay to determine the K of the antibody described herein for one or more peptides. D, the one or more peptides present the ITGA11 epitope in a conformation similar to the epitope in the native protein. The kit can contain, for example, a microtiter plate (the wells of which have been previously conjugated to avidin), and can contain a library of ITGA11-derived peptides, each conjugated to a biotin moiety. Such kits can optionally contain a secondary antibody that specifically binds to the Fc region of the antibodies or antigen-binding fragments thereof described herein, and the secondary antibody can be conjugated to an enzyme that catalyzes a chemical reaction resulting in luminescence (e.g., horseradish peroxidase).

[0269] The kits described herein can also contain the antibodies or antigen-binding fragments thereof described herein and reagents that can be conjugated to such antibodies, including those previously described (e.g., cytotoxic agents, fluorescent molecules, bioluminescent molecules, molecules containing radioisotopes, molecules containing chelating groups that bind to paramagnetic ions, etc.). These kits can additionally contain instructions on how to conjugate the antibodies or antigen-binding fragments thereof described herein to a second molecule (e.g., those molecules described above).

[0270] The kits described herein can also contain a vector that contains a polynucleotide encoding an antibody or an antigen-binding fragment thereof, such as any of the vectors described herein. Optionally, the kit can contain mammalian cells (e.g., CHO cells) that have been genetically engineered to express and secrete an antibody or an antigen-binding fragment thereof or a fragment thereof from the nuclear genome. Such kits can also contain instructions on how to induce the expression of the antibody or an antigen-binding fragment thereof from the polynucleotide, and can additionally contain reagents that can be used to facilitate the transcription of these polynucleotides (e.g., doxycycline or tetracycline). Such kits can be used to produce the antibodies or antigen-binding fragments thereof described herein.

[0271] Other kits described herein can contain tools for engineering prokaryotic or eukaryotic cells (e.g., CHO cells or BL21(DE3) Escherichia coli cells) to express and secrete the antibodies or antigen-binding fragments thereof described herein from the nuclear genome of the cells. For example, the kit can contain CHO cells stored in an appropriate medium and optionally frozen according to methods known in the art. The kit can also provide nucleic acids encoding nucleases (e.g., such as CRISPER / Cas, zinc finger nucleases, TALEN, ARCUS described herein) TMVectors of polynucleotides encoding nucleases and reagents for expressing nucleases in cells. The kit may additionally provide tools for modifying the polynucleotide encoding the nuclease so as to be able to alter the DNA sequence of the nuclease, thereby directing cleavage of a specific target DNA sequence of interest. Examples of such tools include primers for amplification and site-directed mutagenesis of the polynucleotide encoding the nuclease of interest. The kit may also include restriction enzymes, which can be used to selectively excise the polynucleotide encoding the nuclease from the vector and then, after the user has modified the gene, reintroduce the modified polynucleotide into the vector. Such kits may also contain DNA ligases, which can be used to catalyze the formation of covalent phosphodiester bonds between the modified nuclease-encoding polynucleotide and the target vector. The kits described herein may also provide polynucleotides encoding antibodies or antigen-binding fragments thereof, as well as package inserts describing methods that can be used to selectively cleave a specific DNA sequence in the genome of a cell so as to incorporate the polynucleotide encoding the antibody or antigen-binding fragment thereof at that site in the genome. Optionally, the kit may provide polynucleotides encoding fusion proteins that contain an antibody or antigen-binding fragment thereof or a fragment thereof and an additional polypeptide, such as those described herein. Examples

[0272] The following examples are provided to give those of ordinary skill in the art a description of how to use, prepare, and evaluate the compositions and methods described herein, and these examples are intended only as illustrations of the disclosure and are not intended to limit the scope that the inventors regard as their disclosure.

[0273] Example 1. Generation and Purification of Anti-ITGA11 Antibody

[0274] Anti-ITGA11 antibodies were generated using a natural human scFv library with 10^10 unique clone diversities generated by IONTAS. The library was subjected to two rounds of phage display selection, a total of 8 selection rounds, including selections of different combinations of recombinant soluble human ITGA11B1, recombinant murine ITGA11B1, cell-expressed human ITGA11B1, and cell-expressed murine ITGA11B1. Also included was the deselection of cell-expressed human ITGA2B1 and recombinant ITGA2B1. Each selection round was carried out in the presence or absence of 1 mM MnCl2, a total of 8 selection rounds, including outputs with κ light chains and 8 outputs with λ light chains. After the final round of selection and deselection of cell-expressed ITGA11, all selection outputs showed enrichment after the second round of selection.

[0275] After separating the scFv phage display selection output, it is converted to the mammalian-displayed human IgG1 form. These converted outputs are grouped into 4 mammalian-display libraries, and the library sizes are determined using methods known in the art. These libraries are generated for mammalian display transfection and selection in HEK293 cells. One hundred million HEK293 cells are transfected with a 20 μg library DNA mixture, and one day after transfection, the cells from each transfection are 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 transfected libraries are sorted by MACS (Militenyi Midi MACS separator) to enrich cells expressing high levels of human IgG1 (which is detected by anti-PE magnetic beads that bind to the anti-Fc-PE that binds to cells expressing human IgG1). The results for each mammalian display library show enrichment of human ITGA11B1 binders. In round 1 MCD, all MCD libraries are incubated with 0.1 nM biotinylated human ITGA11B1 and sorted using FACS. Unlabeled human ITGA10B1 is included to reduce the number of sorted clones binding to it. Each library is sorted for 30 million cells. The top 1.5% to 4.9% of the human ITGA11B1 binders at 0.1 nM are collected and placed in cell culture for a second round of sorting. This process is repeated in the second round of sorting, increasing the stringency of human ITGA11 binding to enrich for higher-affinity binders and including biotinylated ITGA10 for specific sorting.

[0276] 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 of the output population expression vectors were prepared using methods known in the art and grown in E. coli. Colonies were picked from all outputs and distributed into 1.2 ml of LB medium and kanamycin in a 10x96 deep well plate and incubated overnight with shaking at 37 °C. Each plate contained 90 antibody clones, positive control and negative control antibodies, and 2 blank wells. Plasmid DNA from each well of all 10 plates was extracted and purified using the MagBind kit. The purified plasmid DNA was used to transfect ExpiHEK293 cells for the expression of each soluble antibody clone. The conditioned medium containing each antibody was used for the primary screening experiment. The supernatants from all wells containing human IgG4 antibody clones from 10 plates were evaluated for binding to CHO cells expressing human ITGA11B1 and recombinant human ITGA11B1 protein by ELISA. Among 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 a 24 deep well plate, and the antibodies were purified using protein A affinity chromatography according to standard methods known to those skilled in the art.

[0277] Then, a hierarchical flow scheme was used to functionally evaluate the 61 purified antibody hits, where their ability to bind to cells expressing human ITGA11B1 at 34 nM and their ability to block the binding of human ITGA11B1 to collagen at 68 nM were measured. Among the 61 hits, twenty antibodies were identified that blocked the binding of collagen I 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 out of 20 antibodies bound to murine ITGA11B1.

[0278] Example 2. Binding Characterization of Anti-ITGA11 Antibodies

[0279] Antibody binding to C2C12 cells expressing human ITGA11 or CHO cells expressing murine ITGA11 was performed by incubating the cells with 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 evaluated by flow cytometry. 226 antibodies were screened, and the binding characteristics of 10 antibodies are shown below. The results are provided in Table 2 and are expressed as mean fluorescence intensity (MFI).

[0280] Table 2. Relative binding of human anti-ITGA11 antibodies to human ITGA11 and murine ITGA11

[0281] Antibody Binding to human ITGA11 (MFI) Binding to mouse ITGA11 (MFI) FIB-918-1 3506 2555 FIB-918-6 7885 9407 FIB-918-3 5225 1519 FIB-918-2 30386 12584 FIB-918-5 4271 2744 FIB-918-4 15726 4566 FIB-918-9 1452 2030 FIB-918-10 10567 8572 FIB-918-8 41466 32906 FIB-918-7 7868 21272 Control IgG4 557 513

[0282] Example 3. Anti-ITGA11 antibodies inhibit the binding of collagen to ITGA11 expressed by cells

[0283] Collagen conjugated to fluorescent beads was incubated with anti-ITGA11 antibody and murine C2C12 cells expressing human ITGA11 in binding medium containing DMEM, 10% normal goat serum, and 2% normal rabbit serum for 1 hour. The cells were trypsinized, and unbound collagen (type I rat tail collagen, Corning Labs)-coated beads (fluorescent YG carboxylated microspheres 2 µm (Polyscienced)) were rinsed from the cells 2 times in PBS, 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. Figure 1 、 Figure 2 and Figure 3 The results shown are expressed as the percentage of bound beads relative to the total number of beads incubated with the cells. MOPC 21 mAb IgG1 was used as a negative control.

[0284] Based on the observed IC50 ranges, 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 the binding of collagen to ITGA11 expressed in cells to varying degrees.

[0285] Other embodiments

[0286] The present disclosure has been described in connection with its specific embodiments, and it should be understood that the present disclosure can be further modified, and this application is intended to cover any variations, uses, or adaptations of the present disclosure that generally follow the principles of the present disclosure, and including such departures from the present disclosure within the known or customary scope in the field to which the present disclosure pertains, and can be applied to the basic features set forth above, and follow the scope of the claims. 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), wherein the antibody or antigen-binding fragment thereof comprises: Complementary determining region (CDR)-heavy chain 1 (CDR-H1), which comprises the amino acid sequence 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); Complementary determining region (CDR)-heavy chain 2 (CDR-H2), which comprises the amino acid sequence ISGSGGST (SEQ ID NO:2), ISSSSSTI (SEQ ID NO:15), FDPEDGET (SEQ ID NO:29), or ISGSGGSI (SEQ ID NO:74); Complementary determining region (CDR)-heavy chain 3 (CDR-H3), which comprises the amino acid sequence 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); Complementary determining region (CDR)-light chain 1 (CDR-L1), which comprises the amino acid sequence QSISSY (SEQ ID NO:4), QTIGSY (SEQ ID NO:21), SGSIASNY (SEQ ID NO:31), QGINDF (SEQ ID NO:40), QSVSSSY (SEQ ID NO:49); Complementary determining region (CDR)-light chain 2 (CDR-L2), which comprises the amino acid sequence AAS (SEQ ID NO:5), GAS (SEQ ID NO:22), or EDK (SEQ ID NO:32); and Complementary determining region (CDR)-light chain 3 (CDR-L3), which comprises the amino acid sequence 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).

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

3. The antibody or antigen-binding fragment thereof according to claim 1 or 2, wherein the 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 1-3, wherein the CDR-L1 comprises the amino acid sequence QSISSY (SEQ ID NO:4).

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

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

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

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

8.

9. The antibody or antigen-binding fragment thereof according to claim 8, wherein the antibody or antigen-binding fragment 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.

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

11. The antibody or antigen-binding fragment thereof according to claim 10, wherein the antibody or antigen-binding fragment thereof comprises: A heavy chain variable domain comprising an amino acid sequence having at least 90% sequence identity with 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 with the amino acid sequence of SEQ ID NO:

13.

12. The antibody or antigen-binding fragment thereof according to claim 11, wherein 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.

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

14. The antibody or antigen-binding fragment thereof according to claim 10, wherein the antibody or antigen-binding fragment thereof comprises: A heavy chain variable domain comprising an amino acid sequence having at least 90% sequence identity with 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 with the amino acid sequence of SEQ ID NO:

13.

15. The antibody or antigen-binding fragment thereof according to claim 11, wherein 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.

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

17. The antibody or antigen-binding fragment thereof according to claim 16, wherein the antibody or antigen-binding fragment thereof comprises: A heavy chain variable domain comprising an amino acid sequence having at least 90% sequence identity with 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 with the amino acid sequence of SEQ ID NO:

24.

18. The antibody or antigen-binding fragment thereof according to claim 15, wherein 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.

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

20. The antibody or antigen-binding fragment thereof according to claim 19, wherein the antibody or antigen-binding fragment thereof comprises: A heavy chain variable domain comprising an amino acid sequence having at least 90% sequence identity with 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 with the amino acid sequence of SEQ ID NO:

27.

21. The antibody or antigen-binding fragment thereof according to claim 20, wherein 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.

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

23. The antibody or antigen-binding fragment thereof according to claim 22, wherein the antibody or antigen-binding fragment thereof comprises: A heavy chain variable domain comprising an amino acid sequence having at least 90% sequence identity with 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 with the amino acid sequence of SEQ ID NO:

35.

24. The antibody or antigen-binding fragment thereof according to claim 23, wherein 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.

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

26. The antibody or antigen-binding fragment thereof according to claim 25, wherein the antibody or antigen-binding fragment thereof comprises: A heavy chain variable domain comprising an amino acid sequence having at least 90% sequence identity with 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 with the amino acid sequence of SEQ ID NO:

39.

27. The antibody or antigen-binding fragment thereof according to claim 26, wherein 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.

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

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

42.

30. The antibody or antigen-binding fragment thereof according to claim 29, wherein 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.

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

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

45.

33. The antibody or antigen-binding fragment thereof according to claim 32, wherein 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.

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

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

52.

36. The antibody or antigen-binding fragment thereof according to claim 35, wherein 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.

37. The antibody or antigen-binding fragment thereof according to claim 1, wherein the antibody or antigen-binding fragment thereof comprises a heavy chain variable domain, and the heavy chain variable domain comprises an amino acid sequence having at least 90% sequence identity with 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 according to claim 1 or 36, wherein the antibody or antigen-binding fragment thereof comprises a light chain variable domain, and the light chain variable domain comprises an amino acid sequence having at least 90% sequence identity with any one of the amino acid sequences of SEQ ID NO:8, 13, 19, 24, 35, 39, 42, 45 or 52.

39. The antibody or antigen-binding fragment thereof according to any one of claims 1-38, wherein the antibody or antigen-binding fragment thereof is selected from the group consisting of: monoclonal antibody or antigen-binding fragment thereof, polyclonal antibody or antigen-binding fragment thereof, human antibody or antigen-binding fragment thereof, humanized antibody or antigen-binding fragment thereof, primatized antibody or antigen-binding fragment thereof, bispecific antibody or antigen-binding fragment thereof, multispecific antibody or antigen-binding fragment thereof, dual variable immunoglobulin domain, monovalent antibody or antigen-binding fragment thereof, chimeric antibody or antigen-binding fragment thereof, single-chain Fv molecule (scFv), diabody, triabody, nanobody, antibody-like protein scaffold, domain antibody, Fv fragment, Fab fragment, F(ab’)2 molecule, and tandem scFv (taFv).

40. The antibody or antigen-binding fragment thereof according to claim 39, wherein the antibody or antigen-binding fragment thereof is a human antibody, humanized antibody, or chimeric antibody or antigen-binding fragment thereof.

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

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

43. A vector comprising the polynucleotide according to claim 42.

44. The vector according to claim 43, wherein the vector is an expression vector.

45. The vector according to claim 43, wherein the expression vector is a eukaryotic expression vector.

46. The vector according to claim 45, wherein the vector is a viral vector.

47. The vector according to claim 46, wherein the viral vector is selected from 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 43-47.

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

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

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

52. The method according to claim 51, wherein the disease is a fibrotic disease.

53. The method according to claim 52, wherein the fibrotic disease is selected from hepatic fibrosis, renal fibrosis, skin fibrosis, cardiac fibrosis, vascular fibrosis, ocular fibrosis, fibrosis of the bone marrow, pulmonary fibrosis, glomerulonephritis, heart failure, scleroderma, excessive scar tissue formation after surgery or device insertion, trauma or burns, progressive kidney disease, heart valve disease, hypertensive heart disease, joint and perijoint fibrosis, myelofibrosis, ocular / vitreous fibrosis, intestinal fibrosis and stricture, peritoneal and retroperitoneal fibrosis, pancreatic fibrosis, nephrogenic systemic fibrosis, and primary sclerosing cholangitis.

54. The method according to claim 51, wherein the disease is an inflammatory disease.

55. The method according to claim 54, wherein the inflammatory disease is selected from asthma, airway inflammation, airway hyperreactivity, airway hyperresponsiveness, sinusitis, sinusitis with polyps, nasal polyposis, arthritis, eosinophilic inflammation, mast cell-mediated inflammatory diseases, sepsis, septic shock, seronegative enthesopathy and arthropathy (SEA) syndrome, osteoporosis, eosinophilic esophagitis, scleroderma, dermatitis, atopic dermatitis, allergic rhinitis, bullous pemphigoid, urticaria, cartilage inflammation, polymyalgia rheumatica, polyarteritis nodosa, Wegener's granulomatosis, Behcet's disease, myositis, polymyositis, dermatomyositis, vasculitis, arteritis, diabetic nephropathy, interstitial cystitis, graft-versus-host disease (GVHD), gastrointestinal inflammatory conditions, and inflammatory lung conditions.

56. The method according to claim 51, wherein the disease is cancer.

57. The method according to claim 56, wherein the cancer is selected from breast cancer, colorectal cancer, liver cancer, kidney cancer, hepatocellular carcinoma, lung cancer, pancreatic cancer, gastrointestinal cancer, melanoma, ovarian cancer, prostate cancer, cervical cancer, bladder cancer, glioblastoma, head and neck cancer, and cholangiocarcinoma.

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