Monoclonal antibody against cadherin 11, and method of use.

Abstract

The present invention provides antibodies that bind to cadherin 11 (CDH11) and their antigen-binding fragments, as well as nucleic acids encoding such antibodies and their antigen-binding fragments, and vectors containing such nucleic acids. Further provided are methods and devices for employing such antibodies and / or antigen-binding fragments, including, but not limited to, methods for inhibiting cell-to-cell CDH11-CDH11 interactions and methods for treating diseases in subjects where such intervention is needed. TIFF2026515695000030.tif59149

Description

[Technical Field]

[0001] Cross-reference of related applications This application claims priority to U.S. Patent Application No. 63 / 494,845, filed on 7 April 2023, which is incorporated herein by reference in its entirety.

[0002] Reference to electronic sequence listings This application includes a sequence listing submitted electronically in XML format, which is incorporated herein by reference in its entirety. A copy of the XML was created on 2 April 2024, named SeqList-084284-00294.xml, and is 122,586 bytes in size.

[0003] field This disclosure relates, in general terms, to antibodies against cadherin 11 (CDH11) and methods of using the same. [Background technology]

[0004] background Cadherin 11 (CDH11) is part of the cadherin superfamily, which are membrane proteins involved in homodimeric cell-to-cell interactions. Human CDH11 is a Ca+-dependent transmembrane protein that exhibits homotyped binding. This 796-amino acid protein (approximately 88 kDa) contains five extracellular cadherin domains (approximately 110 aa) named EC1-EC5, a transmembrane region, and a cytoplasmic tail. Human CDH11 shares high homology (approximately 97%) with mouse CDH11 and moderate homology (40-60%) with other type 2 cadherins, with increased sequence conservation in the cytoplasmic tail and EC1-EC2 domains.

[0005] CDH11 is expressed on mesenchymal-derived cells, including hepatic stellate cells (HSCs), myofibroblasts, and fibroblast-like synovial cells (FLS). CDH11 binding increases the production and deposition of matrix proteins, including collagen, leading to fibrosis. CDH11 binding also increases signaling through IL-6 production; MMP1 / MMP2 / MMP3; β-catenin, MAPK (JNK / ERK / p38), RhoA / ROCK, and NFkβ.

[0006] CDH11 has been linked to various fibrotic disorders, including hepatic fibrosis, pulmonary fibrosis, and scleroderma, as well as to rheumatoid arthritis, inflammatory bowel disease, and cancer. There is an urgent need for improved and effective means to combat these diseases by targeting CDH11. [Overview of the project]

[0007] overview Antibodies that bind to CDH11 and their antigen-binding fragments, as well as methods for using such antibodies and their antigen-binding fragments for the treatment of a disease, are provided herein.

[0008] In some embodiments, an anti-cadherin 11 (CDH11) antibody or a fragment thereof that binds to CDH11 is provided, the anti-CDH11 antibody or the fragment thereof that binds to CDH11 comprises a heavy chain variable region and a light chain variable region, each of the heavy chain and light chain variable regions comprising CDR1, CDR2, and CDR3, (a) CDR1H includes SEQ ID NO:1, CDR2H includes SEQ ID NO:11, CDR3H includes SEQ ID NO:20 or SEQ ID NO:21, CDR1L includes SEQ ID NO:35, CDR2L includes SEQ ID NO:43, and CDR3L includes SEQ ID NO:48 or SEQ ID NO:49; (b) CDR1H includes SEQ ID NO:4, CDR2H includes SEQ ID NO:13, CDR3H includes SEQ ID NO:25 or SEQ ID NO:26, CDR1L includes SEQ ID NO:38, CDR2L includes SEQ ID NO:45, and CDR3L includes SEQ ID NO:53; (c) CDR1H includes SEQ ID NO:8, CDR2H includes SEQ ID NO:17, CDR3H includes SEQ ID NO:32, CDR1L includes SEQ ID NO:38, CDR2L includes SEQ ID NO:45, and CDR3L includes SEQ ID NO:59; (d) CDR1H includes SEQ ID NO:2, CDR2H includes SEQ ID NO:11, CDR3H includes SEQ ID NO:22, CDR1L includes SEQ ID NO:36, CDR2L includes SEQ ID NO:43, and CDR3L includes SEQ ID NO:50; (e) CDR1H includes SEQ ID NO:3, CDR2H includes SEQ ID NO:12, CDR3H includes SEQ ID NO:23, CDR1L includes SEQ ID NO:37, CDR2L includes SEQ ID NO:44, and CDR3L includes SEQ ID NO:51; (a) CDR1H includes SEQ ID NO:3, CDR2H includes SEQ ID NO:12, CDR3H includes SEQ ID NO:24, CDR1L includes SEQ ID NO:35, CDR2L includes SEQ ID NO:43, and CDR3L includes SEQ ID NO:52; (b) CDR1H includes SEQ ID NO: 5, CDR2H includes SEQ ID NO: 14, CDR3H includes SEQ ID NO: 27, CDR1L includes SEQ ID NO: 39, CDR2L includes SEQ ID NO: 46, and CDR3L includes SEQ ID NO: 54; (c) CDR1H contains SEQ ID NO:5, CDR2H contains SEQ ID NO:14, CDR3H contains SEQ ID NO:28, CDR1L contains SEQ ID NO:35, CDR2L contains SEQ ID NO:43, and CDR3L contains SEQ ID NO:55; (d) CDR1H contains SEQ ID NO:6, CDR2H contains SEQ ID NO:15, CDR3H contains SEQ ID NO:29, CDR1L contains SEQ ID NO:40, CDR2L contains SEQ ID NO:45, and CDR3L contains SEQ ID NO:56; (e) CDR1H contains SEQ ID NO:7, CDR2H contains SEQ ID NO:16, CDR3H contains SEQ ID NO:30, CDR1L contains SEQ ID NO:41, CDR2L contains SEQ ID NO:44, and CDR3L contains SEQ ID NO:57; (f) CDR1H contains SEQ ID NO:7, CDR2H contains SEQ ID NO:16, CDR3H contains SEQ ID NO:31, CDR1L contains SEQ ID NO:35, CDR2L contains SEQ ID NO:43, and CDR3L contains SEQ ID NO:58; (g) CDR1H contains SEQ ID NO:9, CDR2H contains SEQ ID NO:18, CDR3H contains SEQ ID NO:33, CDR1L contains SEQ ID NO:41, CDR2L contains SEQ ID NO:45, and CDR3L contains SEQ ID NO:60; or (h) CDR1H contains SEQ ID NO:10, CDR2H contains SEQ ID NO:19, CDR3H contains SEQ ID NO:34, CDR1L contains SEQ ID NO:42, CDR2L contains SEQ ID NO:47, and CDR3L contains SEQ ID NO:61.

[0009] In some embodiments, an anti-CDH11 antibody or a fragment thereof that binds to CDH11 is provided, the anti-CDH11 antibody or the fragment thereof that binds to CDH11 comprises a heavy chain variable region and a light chain variable region, each of the heavy chain and light chain variable regions comprising CDR1, CDR2, and CDR3, (a) CDR1H includes SEQ ID NO:1, CDR2H includes SEQ ID NO:11, CDR3H includes SEQ ID NO:20, CDR1L includes SEQ ID NO:35, CDR2L includes SEQ ID NO:43, and CDR3L includes SEQ ID NO:48; (b) CDR1H includes SEQ ID NO:4, CDR2H includes SEQ ID NO:13, CDR3H includes SEQ ID NO:26, CDR1L includes SEQ ID NO:38, CDR2L includes SEQ ID NO:45, and CDR3L includes SEQ ID NO:53; (c) CDR1H includes SEQ ID NO:1, CDR2H includes SEQ ID NO:11, CDR3H includes SEQ ID NO:21, CDR1L includes SEQ ID NO:35, CDR2L includes SEQ ID NO:43, and CDR3L includes SEQ ID NO:49; (d) CDR1H includes SEQ ID NO:4, CDR2H includes SEQ ID NO:13, CDR3H includes SEQ ID NO:25, CDR1L includes SEQ ID NO:38, CDR2L includes SEQ ID NO:45, and CDR3L includes SEQ ID NO:53; or.

[0010] In some embodiments, an anti-CDH11 antibody or a fragment thereof that binds to CDH11 is provided, and the fragment thereof that binds to the anti-CDH11 antibody or CDH11 is as follows: (a) A heavy chain variable region containing at least 80% identical sequences to SEQ ID NO:62, and a light chain variable region containing at least 80% identical sequences to SEQ ID NO:78; (b) A heavy chain variable region containing at least 80% identical sequences to SEQ ID NO:68, and a light chain variable region containing at least 80% identical sequences to SEQ ID NO:83; (c) Heavy chain variable region containing at least 80% identical sequences to SEQ ID NO:75, and light chain variable region containing at least 80% identical sequences to SEQ ID NO:90; (d) A heavy chain variable region containing at least 80% identical sequences to SEQ ID NO:63, and a light chain variable region containing at least 80% identical sequences to SEQ ID NO:79; (e) A heavy chain variable region containing at least 80% identical sequences to SEQ ID NO:64, and a light chain variable region containing at least 80% identical sequences to SEQ ID NO:80; (f) A heavy chain variable region containing at least 80% identical sequences to SEQ ID NO:65, and a light chain variable region containing at least 80% identical sequences to SEQ ID NO:81; (g) A heavy chain variable region containing at least 80% identical sequences to SEQ ID NO:66, and a light chain variable region containing at least 80% identical sequences to SEQ ID NO:82; (h) Heavy chain variable region containing at least 80% identical sequences to SEQ ID NO:67, and light chain variable region containing at least 80% identical sequences to SEQ ID NO:83; (i) A heavy chain variable region containing at least 80% identical sequences to SEQ ID NO:69, and a light chain variable region containing at least 80% identical sequences to SEQ ID NO:84; (j) A heavy chain variable region containing at least 80% identical sequences to SEQ ID NO:70, and a light chain variable region containing at least 80% identical sequences to SEQ ID NO:85; (k) A heavy chain variable region containing at least 80% identical sequences to SEQ ID NO:71, and a light chain variable region containing at least 80% identical sequences to SEQ ID NO:86; (l) A heavy chain variable region containing at least 80% identical sequences to SEQ ID NO:72, and a light chain variable region containing at least 80% identical sequences to SEQ ID NO:87; (m) Heavy chain variable region containing at least 80% identical sequences to SEQ ID NO:73, and light chain variable region containing at least 80% identical sequences to SEQ ID NO:88; (n) A heavy chain variable region containing at least 80% identical sequences to SEQ ID NO:74, and a light chain variable region containing at least 80% identical sequences to SEQ ID NO:89; (o) A heavy chain variable region containing at least 80% identical sequences to SEQ ID NO:76, and a light chain variable region containing at least 80% identical sequences to SEQ ID NO:91; (p) A heavy chain variable region containing at least 80% identical sequences to SEQ ID NO:76, and a light chain variable region containing at least 80% identical sequences to SEQ ID NO:92; or (q) Heavy chain variable region containing at least 80% identical sequences to SEQ ID NO:77, and light chain variable region containing at least 80% identical sequences to SEQ ID NO:93 Includes.

[0011] In some embodiments, a fragment that binds to an anti-CDH11 antibody or CDH11 is provided, and the fragment that binds to the anti-CDH11 antibody or CDH11 is as follows: (a) A heavy chain variable region containing at least 90% identical sequences to SEQ ID NO:62, and a light chain variable region containing at least 90% identical sequences to SEQ ID NO:78; (b) A heavy chain variable region containing at least 90% identical sequences to SEQ ID NO:68, and a light chain variable region containing at least 90% identical sequences to SEQ ID NO:83; (c) Heavy chain variable region containing at least 90% identical sequences to SEQ ID NO:75, and light chain variable region containing at least 90% identical sequences to SEQ ID NO:90; (d) A heavy chain variable region containing at least 90% identical sequences to SEQ ID NO:63, and a light chain variable region containing at least 90% identical sequences to SEQ ID NO:79; (e) A heavy chain variable region containing at least 90% identical sequences to SEQ ID NO:64, and a light chain variable region containing at least 90% identical sequences to SEQ ID NO:80; (f) A heavy chain variable region containing at least 90% identical sequences to SEQ ID NO:65, and a light chain variable region containing at least 90% identical sequences to SEQ ID NO:81; (g) A heavy chain variable region containing at least 90% identical sequences to SEQ ID NO:66, and a light chain variable region containing at least 90% identical sequences to SEQ ID NO:82; (h) A heavy chain variable region containing at least 90% identical sequences to SEQ ID NO:67, and a light chain variable region containing at least 90% identical sequences to SEQ ID NO:83; (i) A heavy chain variable region containing at least 90% identical sequences to SEQ ID NO:69, and a light chain variable region containing at least 90% identical sequences to SEQ ID NO:84; (j) A heavy chain variable region containing at least 90% identical sequences to SEQ ID NO:70, and a light chain variable region containing at least 90% identical sequences to SEQ ID NO:85; (k) A heavy chain variable region containing at least 90% identical sequences to SEQ ID NO:71, and a light chain variable region containing at least 90% identical sequences to SEQ ID NO:86; (l) A heavy chain variable region containing at least 90% identical sequences to SEQ ID NO:72, and a light chain variable region containing at least 90% identical sequences to SEQ ID NO:87; (m) A heavy chain variable region containing at least 90% identical sequences to SEQ ID NO:73, and a light chain variable region containing at least 90% identical sequences to SEQ ID NO:88; (n) A heavy chain variable region containing at least 90% identical sequences to SEQ ID NO:74, and a light chain variable region containing at least 90% identical sequences to SEQ ID NO:89; (o) A heavy chain variable region containing at least 90% identical sequences to SEQ ID NO:76, and a light chain variable region containing at least 90% identical sequences to SEQ ID NO:91; (p) A heavy chain variable region containing at least 90% identical sequences to SEQ ID NO:76, and a light chain variable region containing at least 90% identical sequences to SEQ ID NO:92; or (q) Heavy chain variable region containing at least 90% identical sequences to SEQ ID NO:77, and light chain variable region containing at least 90% identical sequences to SEQ ID NO:93 Includes.

[0012] In some embodiments, a fragment that binds to an anti-CDH11 antibody or CDH11 is provided, and the fragment that binds to the anti-CDH11 antibody or CDH11 is as follows: (a) Heavy chain variable region including SEQ ID NO:62, and light chain variable region including SEQ ID NO:78; (b) Heavy chain variable region including SEQ ID NO:68, and light chain variable region including SEQ ID NO:83; (c) Heavy chain variable region including SEQ ID NO:75, and light chain variable region including SEQ ID NO:90; (d) Heavy chain variable region including SEQ ID NO:63, and light chain variable region including SEQ ID NO:79; (e) Heavy chain variable region including SEQ ID NO:64, and light chain variable region including SEQ ID NO:80; (f) Heavy chain variable region including SEQ ID NO:65, and light chain variable region including SEQ ID NO:81; (g) Heavy chain variable region including SEQ ID NO:66, and light chain variable region including SEQ ID NO:82; (h) Heavy chain variable region including SEQ ID NO:67, and light chain variable region including SEQ ID NO:83; (i) Heavy chain variable region including SEQ ID NO:69, and light chain variable region including SEQ ID NO:84; (j) Heavy chain variable region including SEQ ID NO:70, and light chain variable region including SEQ ID NO:85; (k) Heavy chain variable region including SEQ ID NO:71, and light chain variable region including SEQ ID NO:86; (l) Heavy chain variable region including SEQ ID NO:72, and light chain variable region including SEQ ID NO:87; (m) Heavy chain variable region including SEQ ID NO:73, and light chain variable region including SEQ ID NO:88; (n) Heavy chain variable region including SEQ ID NO:74, and light chain variable region including SEQ ID NO:89; (o) Heavy chain variable region including SEQ ID NO:76, and light chain variable region including SEQ ID NO:91; (p) Heavy chain variable region including SEQ ID NO:76, and light chain variable region including SEQ ID NO:92; or (q) Heavy chain variable region including SEQ ID NO:77, and light chain variable region including SEQ ID NO:93 Includes.

[0013] In one embodiment, the CDH11 antibody or the fragment that binds to CDH11 is a monoclonal antibody. In one embodiment, the CDH11 antibody or the fragment that binds to CDH11 is a recombinant antibody. In one embodiment, the CDH11 antibody or the fragment that binds to CDH11 includes a human framework region or a modified human framework region. In one embodiment, the CDH11 antibody or the fragment that binds to CDH11 includes a human constant region or a modified human constant region. In one embodiment, the fragment that binds to CDH11 is scFv, Fv, Fab', Fab, F(ab')2, or a diabody. In one embodiment, the anti-CDH11 antibody or the fragment that binds to CDH11 is deglycosylated. In several embodiments, the anti-CDH11 antibody or the fragment that binds to CDH11 is conjugated with one or more of the following: cytotoxicity, fluorescent labeling, and imaging agents.

[0014] Provided herein are nucleic acid molecules encoding an anti-CDH11 antibody or a fragment thereof that binds to CDH11. In some embodiments, the nucleic acid molecule is as follows: (a) SEQ ID NO: 94 and / or SEQ ID NO: 111; (b) SEQ ID NO: 95 and / or SEQ ID NO: 112; (c) SEQ ID NO:96 and / or SEQ ID NO:113; (d) SEQ ID NO:97 and / or SEQ ID NO:114; (e) SEQ ID NO:98 and / or SEQ ID NO:115; (f) SEQ ID NO: 99 and / or SEQ ID NO: 116; (g) SEQ ID NO: 100 and / or SEQ ID NO: 117; (h) SEQ ID NO: 101 and / or SEQ ID NO: 118; (i) SEQ ID NO: 102 and / or SEQ ID NO: 119; (j) SEQ ID NO: 103 and / or SEQ ID NO: 120; (k) SEQ ID NO: 104 and / or SEQ ID NO: 121; (l) SEQ ID NO: 105 and / or SEQ ID NO: 122; (m) SEQ ID NO: 106 and / or SEQ ID NO: 123; (n) SEQ ID NO: 107 and / or SEQ ID NO: 124; (o) SEQ ID NO: 108 and / or SEQ ID NO: 125; (p) SEQ ID NO: 109 and / or SEQ ID NO: 126; or (q) SEQ ID NO: 110 and / or SEQ ID NO: 127 Includes.

[0015] A vector or set of vectors comprising nucleic acid molecules disclosed herein is provided.

[0016] Isolated host cells containing nucleic acid molecules, vectors, or sets of vectors disclosed herein are provided.

[0017] A method for producing an anti-CDH11 antibody or a fragment thereof that binds to CDH11 is provided, comprising the step of culturing a host cell under conditions that enable the host cell to produce an anti-CDH11 antibody or a fragment thereof that binds to CDH11.

[0018] A pharmaceutical composition is provided comprising an anti-CDH11 antibody or a fragment thereof bound to CDH11, and a pharmaceutically acceptable excipient.

[0019] A method is provided for inhibiting intercellular CDH11-CDH11 interactions, comprising the step of bringing cells into contact with an anti-CDH11 antibody or a fragment thereof that binds to CDH11, or a pharmaceutical composition disclosed herein. A method is also provided for inhibiting intercellular CDH11-CDH11 interactions in a subject where such inhibition is necessary, comprising the step of administering the anti-CDH11 antibody or a fragment thereof that binds to CDH11, or a pharmaceutical composition disclosed herein, to a subject. In one embodiment, the method comprises the step of inhibiting CDH11-CDH11 interactions between hepatic stellate cells (HSCs). A method is also provided for treating fibrosis in a subject where such treatment is necessary, comprising the step of administering the anti-CDH11 antibody or a fragment thereof that binds to CDH11, or a pharmaceutical composition disclosed herein, to a subject. In multiple embodiments, fibrosis is hepatic fibrosis; pulmonary fibrosis; renal fibrosis; cardiac fibrosis; myelofibrosis; pancreatic fibrosis; cutaneous fibrosis, including systemic scleroderma (scleroderma) and keloids; intestinal fibrosis; post-myocardial infarction fibrosis; post-replacement fibrosis; perivascular fibrosis; or articular fibrosis. In one embodiment, fibrosis is hepatic fibrosis. In multiple embodiments, the subject has an infection caused by a hepatitis virus, non-alcoholic fatty liver disease (NASH), a hereditary metabolic disorder, a cholestatic disorder, or an immune disorder. In multiple embodiments, hepatic fibrosis is associated with increased alcohol intake, excessive vitamin A intake, or drug toxicity. In multiple embodiments, hepatic fibrosis is associated with an autoimmune disease of the liver or bile duct, liver graft rejection, or a genetic disorder of the liver or bile duct. In multiple embodiments, the autoimmune disease of the liver or bile duct is autoimmune hepatitis, primary biliary cholangitis, or primary sclerosing cholangitis. In multiple embodiments, the hepatic genetic disorder is Wilson's disease or hemochromatosis, or the hepatic genetic disorder of the bile duct is biliary atresia. In one embodiment, fibrosis is intestinal fibrosis and intestinal stricture associated with colitis. In multiple embodiments, the subject has inflammatory bowel disease (IBD), Crohn's disease, or colitis. In multiple embodiments, pulmonary fibrosis is idiopathic pulmonary fibrosis or induced by toxic injury.In several embodiments, the subject has scleroderma or systemic scleroderma. In one embodiment, the subject has keloid scars. A method is provided for treating rheumatoid arthritis in a subject in need, comprising the step of administering to the subject an anti-CDH11 antibody or a fragment thereof conjugated to CDH11, or a pharmaceutical composition disclosed herein. The subject may be human. [Brief explanation of the drawing]

[0020] [Figure 1A] Figures 1A and 1B illustrate immunization strategies for generating anti-CDH11 antibodies. Figure 1A: Strategy for DNA immunization. Figure 1B: Strategy for protein immunization. [Figure 1B] Refer to the explanation in Figure 1A. [Figure 2A]Figures 2A and 2B show data for the serum screening performed. Mouse serum was collected after each round of immunization and tested by either flow cytometry or ELISA. Figure 2A: For the ELISA experiment, human CDH11 ECD protein, c-terminus tagged with human Fc(IgG1), 5 μg / ml was coated onto an ELISA plate. Serum was added and detected by a secondary antibody (Jackson ImmunoResearch) conjugated with goat anti-mouse IgG(Fc-specific)-horseradish peroxidase (HRP), and colored using ABTS (2,2'-azinobis[3-ethylbenzothiazoline-6 ​​sulfonic acid] diammonium salt) substrate (ThermoFisher). Absorbance of the ELISA plate was read at OD 405 nm. Protein, mouse (see Figure 1B): Left bar = 1st protein boost. Right bar = 2nd + protein boost. DNA-protein, mouse (see Figure 1A): Left bar = 1st protein boost. Right bar = 2nd + protein boost. 135, 137, 138, 139, 141, and 143 refer to antibody clone numbers. RGN NMS = normal mouse serum (unimmunized serum). Figure 2B: For flow cytometry experiments, Expi293F HEK cells were transfected with either full-length human or mouse CDH11, tagged with GFP at the C-terminus. Goat anti-mouse IgG (Fc-specific) conjugated with allophycocyanin (APC, Jackson ImmunoResearch) was used as a secondary antibody at a 1:1000 dilution. Normal mouse serum was included as a negative control. Mouse serum was diluted at 1:100, 1:1000, and 1:10000. Flows for APC fluorescence were reported as mean fluorescence intensity (MFI). Protein, mouse (see Figure 1B): Bars from left to right: Human CDH11 (HuCAD11), 1st protein boost; Human CDH11, 2nd + protein boost; Mouse CDH11 (MsCAD11), 1st protein boost; Mouse CDH11, 2nd + protein boost.DNA + protein, mouse (see Figure 1A): Bars from left to right: Human CDH11, DNA boost; Human CDH11, first protein boost; Mouse CDH11, DNA boost; Mouse CDH11, first protein boost. [Figure 2B] Refer to the explanation in Figure 2A. [Figure 3A-1] Figures 3A and 3B show data for the hybridoma screening performed. Hybridomas were screened against human CDH11 expressed by Expi293F HEK cells, with the C-terminus tagged with GFP (using flow cytometry), or against human CDH11 extracellular domain protein (tagged with human IgG1 Fc tag) (by ELISA). For flow cytometry, hybridomas were detected by a goat anti-mouse IgG Fc-specific-APC conjugate (Jackson ImmunoResearch). ELISA was colorimetrically chromogenically using a goat anti-mouse IgG (Fc-specific)-HRP conjugate (Jackson ImmunoResearch). A total of 47 clones were evaluated. Left bar: Flow MFI (right axis), Right bar: ELISA, absorbance at OD 450 nm (left axis). The hybridoma supernatant was diluted 1:1000 in FACS buffer, and a commercially available control, anti-huCDH11 antibody 16G5 (BioLegend), was added at 1 μg / ml. Figure 3A: Fusions 1 and 2. Figure 3B: Fusion 3. [Figure 3A-2] Refer to the explanation in Figure 3A-1. [Figure 3B] Refer to the explanation in Figure 3A-1. [Figure 4] Figure 4 illustrates the binding of selected hybridoma-derived serum to human and rat primary FLS, as determined by flow cytometry. Left bar: human FLS, right bar: rat FLS. FS = hybrid serum (blood serum obtained from the animal on the day the spleen was isolated to produce the hybridoma). NMS = normal mouse serum (unimmunized serum). [Figure 5]Figure 5 illustrates the cross-reactivity of isolated anti-CDH11 antibodies with different type 2 cadherins. Right bar: Mouse CDH11 (MoCAD11). Anti-CAD11 = Mouse anti-human CAD11 antibody, clone 16G5 (BioLegend), positive control. [Figure 6A] Figures 6A, 6B, 6C, and 6D illustrate the effect of monoclonal anti-human CDH11 antibodies on rat FLS infiltration. Seventeen different monoclonal antibodies specific to human CDH11 were tested for infiltration inhibitory activity in 3-4 FLS cell lines derived from arthritis rats. Eleven different antibodies reduced FLS infiltration (arrows), five of which exhibited a dose-dependent mechanism. For comparison, two different commercially available anti-CDH11 antibodies were used: R&D Systems' goat anti-human CDH11 ("PG") and BioLegend's monoclonal anti-CDH11 antibody clone 16G5 ("TM"). ctl = control (cells not treated with antibody). [Figure 6B] Refer to the explanation in Figure 6A. [Figure 6C] Refer to the explanation in Figure 6A. [Figure 6D] Refer to the explanation in Figure 6A. [Figure 7A] Figures 7A, 7B, 7C, 7D, 7E, 7F, 7G, and 7H illustrate the effects of monoclonal anti-human CDH11 antibodies on RA (human rheumatoid arthritis) FLS infiltration. Twenty-seven different monoclonal antibodies specific to human CDH11 were tested for infiltration inhibitory activity in 3-4 FLS cell lines derived from patients with RA. Seven different antibodies reduced FLS infiltration by at least 30% (arrows), and some showed dose-dependent effects. Figures 7A, 7B, 7C, and 7D: First batch of monoclonal antibodies. Figure 7E: For further confirmation, the positive antibody selected from the first batch (derived from Figure 7D) that showed the most significant FLS infiltration inhibitory effect was tested again on a different FLS cell line (arrows). Figures 7F, 7G, and 7H: Second batch of anti-human CDH11 monoclonal antibodies. CADH-11 = control antibody. ctl = control (cells not treated with antibody). [Figure 7B] Refer to the explanation in Figure 7A. [Figure 7C] Refer to the explanation in Figure 7A. [Figure 7D] Refer to the explanation in Figure 7A. [Figure 7E] Refer to the explanation in Figure 7A. [Figure 7F] Refer to the explanation in Figure 7A. [Figure 7G] Refer to the explanation in Figure 7A. [Figure 7H] Refer to the explanation in Figure 7A. [Figure 8] Figures 8A and 8B illustrate the effects of selected anti-CDH11 antibodies on mouse FLS infiltration. Figure 8A: Two of the anti-CDH11 antibodies reduced the infiltration of FLS from KRN serum-induced arthritis (KSIA) mice (arrows). Figure 8B: Five of the anti-CDH11 antibodies reduced the infiltration of FLS from mice with collagen antibody-induced arthritis (CAIA) (arrows). Data are shown as the average of pairs per treatment state. [Figure 9] Figure 9 illustrates immunofluorescence staining of mouse FLS using selected monoclonal anti-human CDH11 antibodies. FLS from KSIA mice were used for immunofluorescence using either a commercially available anti-CDH11 antibody (16G5, Biolegend, catalog number 368702) or six monoclonal anti-CDH11 antibodies shown to inhibit RA FLS infiltration. Of the six antibodies tested, four (16A10, 2A10, 3A10, and 5H1) stained positively compared to isotype controls, while two (2F6 and 2A1) stained negatively. Left ("anti-CDH11") = Alexa 488 secondary anti-mouse antibody; Right ("anti-CDH11 / actin") = Alexa 488 secondary anti-mouse antibody and Alexa 594 phalloidin (staining actin filaments). [Figure 10]Figure 10 illustrates the in vivo study of selected anti-human CDH11 antibodies in mice with KSIA. Four to five male C57BL / 6 mice per treatment group received 100 μL of KRN arthritis-inducing serum on days 0 and 2. Starting on day 0, mice were started with 10 mg / kg of antibody (IgG1 isotype control, 2A10, 3A10, or 10E1) or PBS per day, every other day for 10 days. Mice were also scored every other day. Mice treated with monoclonal 3A10 had significantly lower arthritis severity scores on day 8 compared to the other treatment groups (compared to the PBS group, P=0.03; unpaired t-test). [Figure 11] Figure 11 shows that the monoclonal anti-CDH11 antibody 3A10 reduced ankle and foot edema in mice with KSIA (representative image). [Figure 12A] Figures 12A, 12B, and 12C illustrate the binding of selected anti-CHH11 antibodies to cellular CDH11. Antibodies 2A10, 3A10, and 10E1 were tested for binding to human or mouse CDH11 expressed on Expi 293F cells. Antibodies were tested sequentially in 1:3 dilutions over eight steps, starting at 33 nM. Binding was measured using standard goat anti-mouse IgG (Fc-specific) conjugated with APC (Jackson ImmunoResearch). A commercially available control antibody, 16G5, was also evaluated as a control. Figure 12A: Antibody binding to human CDH11. Figure 12B: Antibody binding to mouse CDH11. Figure 12C: Antibody binding to human CDH11. Traces at 1 nM are, from top to bottom: 2A10; 3A10; 16G5; 10E1; mAb2 control antibody; mAb1 / isotype control. [Figure 12B] Refer to the explanation in Figure 12A. [Figure 12C] Refer to the explanation in Figure 12A. [Figure 13]Figure 13 illustrates the structure and different domains of human CDH11. Excerpted from Brasch J, Harrison OJ, Honig B, Shapiro L. Thinking outside the cell: how cadherins drive adhesion. Trends Cell Biol. 2012 Jun;22(6):299-310. [Figure 14A] Figures 14A, 14B, and 14C illustrate the results of domain mapping experiments for selected anti-CDH11 antibodies. Five DNA constructs encoding human CDH11 variants (labeled “Domain 1 Missing” to “Domain 5 Missing”) were generated, each CDH11 variant lacking one of the five EC domains. The C-terminus of the CDH11 variants was fused with GFP. The five constructs, or the construct encoding full-length human CDH11, were transfected into Expi293F cells. Antibody binding (1 μg / ml) to cells expressing different CDH11 variants was measured by flow cytometry. A goat anti-mouse IgG (Fc-specific)-APC conjugate secondary antibody (Jackson ImmunoResearch) was used for detection. Results are graphed as mean fluorescence intensity (MFI). Figure 14A: Bars, from left to right: 2A10, 3A10, 10E1, 16G5 (BioLegend, control). Figure 14B: Bars, from left to right: 1A1, 2A1, 2D2, 6D5, 16G5 (control). Figure 14C: Evaluation of fusion serum and normal serum from immunized mice. Bars, from left to right: 16G5 (control), fusion serum, NMS, isotype control. [Figure 14B] Refer to the explanation in Figure 14A. [Figure 14C] Refer to the explanation in Figure 14A. [Figure 15A]Figures 15A, 15B, 15C, 15D, 15E, 15F, 15G, and 15H illustrate the efficacy of anti-CDH11 antibodies in well-established fibrosis models. Figure 15A: Anti-CDH11 antibodies did not affect the survival rates of two hHSC cell lines (LX-2 = immortalized hHSC; TWNT4 = primary cell line established from patients with liver disease). Figure 15B: Anti-CDH11 antibodies significantly reduced the expression of type I collagen (Col1a1), actin α2, smooth muscle (Acta2), platelet-derived growth factor receptor β (bPDGFR), matrix metalloproteinase-1 (MMP1), and matrix metalloproteinase-2 (MMP2) in both LX-2 hHSCs and the primary hHSC line TWNT4. These are genes known to mediate the fibrotic process (*P ≤ 0.05). Figures 15C and 15D: Anti-CDH11 antibodies significantly reduced Col1a1 protein production both by LX-2 and by the primary hHSC strain TWNT4, further demonstrating their anti-fibrotic properties (*P ≤ 0.05). Figure 15E: Anti-CDH11 antibody 3A10 significantly reduced the expression of fibrillating genes in primary hHSCs. Figure 15F: Anti-CDH11 antibody 3A10 significantly reduced Col1a1 production in primary hHSCs. *P < 0.05. Figure 15G: Anti-CDH11 antibody 3A10 reduced proliferation of primary hHSCs. *P < 0.05. Figure 15H: αSMA protein expression was significantly downregulated in hPCLS treated with Ab-3A10 for 48 hours (Patient 1). [Figure 15B-1] Refer to the explanation in Figure 15A. [Figure 15B-2] Refer to the explanation in Figure 15A. [Figure 15B-3] Refer to the explanation in Figure 15A. [Figure 15C] Refer to the explanation in Figure 15A. [Figure 15D] Refer to the explanation in Figure 15A. [Figure 15E-1] Refer to the explanation in Figure 15A. [Figure 15E-2] Refer to the explanation in Figure 15A. [Figure 15F-1] Refer to the explanation in Figure 15A. [Figure 15F-2] Refer to the explanation in Figure 15A. [Figure 15G-1] Refer to the explanation in Figure 15A. [Figure 15G-2] Refer to the explanation in Figure 15A. [Figure 15G-3] Refer to the explanation in Figure 15A. [Figure 15H] Refer to the explanation in Figure 15A. [Figure 16] Figure 16: Anti-CDH11 antibody 3A10 was not cytotoxic to human precisely cut liver slices (hPCLS). [Modes for carrying out the invention]

[0021] Detailed explanation Antibodies that bind to CDH11 and their antigen-binding fragments, as well as methods for using such antibodies and their antigen-binding fragments for the treatment of a disease, are provided herein. As used herein, the terms CAD11, CDH11, and CADH-11 are used interchangeably to mean cadherin 11.

[0022] Fibrosis has been estimated to contribute to 40% of mortality in the population. These conditions include hepatic fibrosis, pulmonary fibrosis, scleroderma, bowel stenosis in inflammatory bowel disease (IBD), post-myocardial infarction fibrosis, and articular fibrosis. Hepatic fibrosis can be caused by several different chronic liver diseases, including common ones such as non-alcoholic steatohepatitis (NASH), increased alcohol intake, hereditary metabolic disorders, drug use, excessive vitamin A intake, cholestatic disorders, and immune disorders. Furthermore, hepatic fibrosis can progress to liver failure and / or liver cancer, reducing survival rates. Despite years of research, there are currently no FDA-approved drugs to treat hepatic fibrosis. CDH11 is primarily expressed in mesenchymal-derived cells, including hematopoietic stem cells (HSCs), in the liver, and its expression is increased in similar mesenchymal cells (liver, lung, skin, colitis with stenosis) in a variety of fibrotic states. Hemolytic stem cells (HSCs) are mesenchymal-derived cells that play a central role in hepatic fibrosis and produce type I collagen and other matrix proteins. These cells can be used to study pro-fibrotic and anti-fibrotic processes in the liver and to screen for new therapeutic agents. mRNA levels of CDH11 are elevated in the plasma of patients with hepatic fibrosis. Furthermore, elevated levels of CDH11 correlate with type I collagen in cholestatic hepatic fibrosis. Compositions and methods for suppressing the fibrogenic phenotype and function of HSCs are provided herein. Compositions and methods for treating fibrotic disorders are also provided herein.

[0023] Rheumatoid arthritis (RA) affects 1% of the population and is associated with an increased risk of birth defects and physical disability, as well as a reduced lifespan. While several treatments for RA have emerged in the past decade, disease remission remains rare, highlighting the need for newer and more effective therapies. Current treatments targeting different aspects of the immune response, such as IL-1, IL-6, TNFα, and JAK, typically achieve only a 20% improvement in disease activity in most patients (60-70%), far from the desired disease remission. These therapies also induce immunosuppression, limiting dose increases and associated with an increased risk of infection. CDH11 is expressed at increased levels in synovial tissue and FLS from arthritis animals compared to non-arthritis animals. The in vitro invasive nature of FLS is significantly correlated with in vivo joint injury. CDH11 is required for the formation and stabilization of the synovial tissue's lining layer, which is central to the development of synovial hyperplasia and articular erosive changes. CDH11 knockout mice are protected from arthritis, and their FLS has reduced infiltrativeness (this feature has been shown to correlate with reduced disease severity and joint injury in RA and in rodent models of RA). CDH11 binding increases IL-6 production by the FLS, synergistically activating NFκB and MAP kinases with TNFα and IL-1β. Compositions and methods for reducing FLS infiltrativeness and / or joint destruction are provided herein. Compositions and methods for reducing the activity and severity of RA disease are also provided herein. Compositions and methods for preventing or reducing joint injury in patients with RA are provided herein. By targeting the disease site (synovial membrane), the compositions and methods disclosed herein enable disease control by targeting the disease site and thus reducing the risk of systemic immunosuppression.

[0024] Inflammatory bowel disease (IBD) affects 6 to 8 million people worldwide. Treatment typically involves the use of various immunosuppressants, but 40% of patients still do not respond to these treatments. Intestinal fibrosis and strictures are major complications of IBD, affecting as many as 30% of patients, especially those with Crohn's disease. Despite advances in IBD treatment, there has been no progress in treating or preventing intestinal fibrosis and strictures, and the incidence of this complication has not changed in the past decade, with treatment options including endoscopic balloon dilation or surgery. The development of intestinal fibrosis and stricture formation involves intestinal mesenchymal cells (stromal cells), including fibroblasts and myofibroblasts. These cells accumulate in the IBD intestine and proliferate in response to inflammatory mediators whose expression levels are increased in IBD, such as TNFα and IL-1β, and then produce excess cellular matrix proteins, including collagen, leading to fibrosis. None of the current IBD treatments and strategies target these intestinal mesenchymal cells. These intestinal mesenchymal cells express CDH11, and its levels are elevated in the intestines of patients with IBD. CDH11-deficient (knockout) mice studied in a TNBS-induced IBD model showed significantly reduced fibrosis and collagen deposition in the subepithelial and subserosa of the intestine compared to wild-type mice expressing CDH11. Compositions and methods for blocking hemophilic interactions of the CHD11 molecule are provided herein to inhibit CDH11-mediated production of collagen, fibronectin, IL-6, and TGFβ in the IBD intestine, thereby reducing the risk of fibrosis and mitigating the progression of established fibrosis. In several embodiments, the methods disclosed herein do not immunosuppress the patient and can therefore be used in combination with standard IBD care treatments, such as anti-TNFα agents and others.

[0025] antibody The term "antibody" is used in its broadest sense and includes monoclonal antibodies (including full-length or intact monoclonal antibodies), polyclonal antibodies, multivalent antibodies, and polyspecific antibodies (e.g., bispecific antibodies).

[0026] As used herein, "antibody variable domain," "variable chain," etc., refer to a portion of the light and heavy chains of an antibody molecule, including the complementarity-determining regions (CDRs; i.e., CDR1, CDR2, and CDR3), as well as the amino acid sequences of the framework region. H This refers to the variable domain of the heavy chain. L This refers to the variable domain of the light chain. The term "framework region" refers to variable domain residues other than CDR residues. Furthermore, the "light chains" of antibodies (immunoglobulins) derived from any vertebrate species can be assigned to one of two distinct types called kappa (κ) and lambda (λ) based on the amino acid sequence of their constant domains.

[0027] As used herein, the term “complementarity-determining region” (CDR) refers to a portion of the antibody variable domain that is (usually) involved in antigen binding. Each variable domain typically has three CDR regions, identified as CDR1, CDR2, and CDR3. Each CDR is defined by amino acid residues from a CDR as defined by Kabat, for example (i.e., approximately residues 24-34 (L1), 50-56 (L2), and 89-97 (L3) in the light chain variable domain, and residues 31-35 (H1), 50-65 (H2), and 95-102 (H3) in the heavy chain variable domain) (Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md. (1987, 1991)) may contain amino acid residues from the "hypervariable loop" (i.e., approximately residues 26-32 (LI), 50-52 (L2), and 91-96 (L3) in the light chain variable domain, and residues 26-32 (H1), 53-55 (H2), and 96-101 (H3) in the heavy chain variable domain) (Chothia & Lesk 196 J. Mol. Biol. 901 (1987)) may also be included. In some cases, the CDR may contain amino acids derived from both the CDR region and the hypervariable loop as defined by Kabat. The designation of Kabat residues does not always directly correspond to the linear numbering of amino acid residues (primary amino acid sequence). The actual linear amino acid sequence may contain fewer or additional amino acids than the strict Kabat numbering, which corresponds to the shortening or insertion of structural elements into the basic variable domain structure, whether it be a framework or a CDR. The correct Kabat numbering of residues can be determined for a given antibody or antigen-binding fragment by aligning homologous residues in the sequence of the antibody or antigen-binding fragment with the "standard" Kabat-numbered sequence.Alternatively, CDR can be defined according to the ImMunoGeneTics (IMGT) system (Lefranc, M.-P. et al., Dev. Comp. Immunol., 27, 55-77 (2003)).

[0028] As used herein, the term “constant region” refers to a region of an immunoglobulin light chain or immunoglobulin heavy chain distinct from the variable region. The constant domain of a heavy chain generally includes at least one of the following: a CH1 domain, hinges (e.g., upper, middle, and / or lower hinge regions), a CH2 domain, and a CH3 domain. For example, the antibodies described herein may include polypeptides containing a CH1 domain; polypeptides containing a CH1 domain, at least a portion of a hinge domain, and a CH2 domain; polypeptides containing a CH1 domain and a CH3 domain; polypeptides containing a CH1 domain, at least a portion of a hinge domain, and a CH3 domain; or polypeptides containing a CH1 domain, at least a portion of a hinge domain, a CH2 domain, and a CH3 domain. In some embodiments, the polypeptide includes a polypeptide chain containing a CH3 domain. The constant domain of a light chain may be a kappa (κ) or lambda (λ) constant region. However, it will be understood by those skilled in the art that these constant domains (e.g., heavy or light chains) can be modified from naturally occurring immunoglobulin molecules to vary in their amino acid sequence. HC is V H And refers to the heavy chain, including the steady region. LC is V L This refers to a light chain that includes a steady-state region.

[0029] As used herein, the terms “Fc region” or “Fc portion” refer to the C-terminal region of an immunoglobulin heavy chain. The Fc region may be a naturally occurring Fc region or a variant Fc region that does not exist naturally. Generally, the Fc region of an immunoglobulin contains the constant domains CH2 and CH3. The boundaries of the Fc region can vary, but in some embodiments, the human IgG heavy chain Fc region can be defined as extending from the amino acid residue at position C226 or from the amino acid residue at position P230 to its carboxyl terminus. In some embodiments, the “CH2 domain” of the human IgG Fc region typically extends from approximately 231 to approximately 340 amino acid residues. In some embodiments, an N-linked glycan is sandwiched between two CH2 domains of an intact natural IgG molecule. In some embodiments, the “CH3 domain” of the human IgG Fc region includes the C-terminal residues relative to the CH2 domain, for example, from approximately 341 to approximately 447 amino acid residues of the Fc region.

[0030] The terms “the antigen-binding fragment” and “the fragment that binds to CDH11” are used interchangeably herein.

[0031] An anti-CDH11 antibody or a fragment thereof that binds to CDH11 is provided herein, the anti-CDH11 antibody or the fragment thereof that binds to CDH11 comprises a heavy chain variable region and a light chain variable region, the heavy chain and light chain variable regions each comprising CDR1, CDR2, and CDR3, where CDR1H comprises SEQ ID NO:1, CDR2H comprises SEQ ID NO:11, CDR3H comprises SEQ ID NO:20 or SEQ ID NO:21, CDR1L comprises SEQ ID NO:35, CDR2L comprises SEQ ID NO:43, and CDR3L comprises SEQ ID NO:48 or SEQ ID NO:49.

[0032] An anti-CDH11 antibody or a fragment thereof that binds to CDH11 is provided herein, the anti-CDH11 antibody or the fragment thereof that binds to CDH11 comprises a heavy chain variable region and a light chain variable region, the heavy chain and light chain variable regions each comprising CDR1, CDR2, and CDR3, where CDR1H comprises SEQ ID NO:1, CDR2H comprises SEQ ID NO:11, CDR3H comprises SEQ ID NO:20, CDR1L comprises SEQ ID NO:35, CDR2L comprises SEQ ID NO:43, and CDR3L comprises SEQ ID NO:48.

[0033] An anti-CDH11 antibody or a fragment thereof that binds to CDH11 is provided herein, the anti-CDH11 antibody or the fragment thereof that binds to CDH11 comprises a heavy chain variable region and a light chain variable region, the heavy chain and light chain variable regions each comprising CDR1, CDR2, and CDR3, where CDR1H comprises SEQ ID NO:1, CDR2H comprises SEQ ID NO:11, CDR3H comprises SEQ ID NO:21, CDR1L comprises SEQ ID NO:35, CDR2L comprises SEQ ID NO:43, and CDR3L comprises SEQ ID NO:49.

[0034] An anti-CDH11 antibody or a fragment thereof that binds to CDH11 is provided herein, the anti-CDH11 antibody or the fragment thereof that binds to CDH11 comprises a heavy chain variable region and a light chain variable region, the heavy chain and light chain variable regions each comprising CDR1, CDR2, and CDR3, where CDR1H comprises SEQ ID NO:4, CDR2H comprises SEQ ID NO:13, CDR3H comprises SEQ ID NO:25 or SEQ ID NO:26, CDR1L comprises SEQ ID NO:38, CDR2L comprises SEQ ID NO:45, and CDR3L comprises SEQ ID NO:53.

[0035] An anti-CDH11 antibody or a fragment thereof that binds to CDH11 is provided herein, the anti-CDH11 antibody or the fragment thereof that binds to CDH11 comprises a heavy chain variable region and a light chain variable region, the heavy chain and light chain variable regions each comprising CDR1, CDR2, and CDR3, where CDR1H comprises SEQ ID NO:4, CDR2H comprises SEQ ID NO:13, CDR3H comprises SEQ ID NO:25, CDR1L comprises SEQ ID NO:38, CDR2L comprises SEQ ID NO:45, and CDR3L comprises SEQ ID NO:53.

[0036] An anti-CDH11 antibody or a fragment thereof that binds to CDH11 is provided herein, the anti-CDH11 antibody or the fragment thereof that binds to CDH11 comprises a heavy chain variable region and a light chain variable region, the heavy chain and light chain variable regions each comprising CDR1, CDR2, and CDR3, where CDR1H comprises SEQ ID NO:4, CDR2H comprises SEQ ID NO:13, CDR3H comprises SEQ ID NO:26, CDR1L comprises SEQ ID NO:38, CDR2L comprises SEQ ID NO:45, and CDR3L comprises SEQ ID NO:53.

[0037] An anti-CDH11 antibody or a fragment thereof that binds to CDH11 is provided herein, the anti-CDH11 antibody or the fragment thereof that binds to CDH11 comprises a heavy chain variable region and a light chain variable region, the heavy chain and light chain variable regions each comprising CDR1, CDR2, and CDR3, where CDR1H comprises SEQ ID NO:8, CDR2H comprises SEQ ID NO:17, CDR3H comprises SEQ ID NO:32, CDR1L comprises SEQ ID NO:38, CDR2L comprises SEQ ID NO:45, and CDR3L comprises SEQ ID NO:59.

[0038] An anti-CDH11 antibody or a fragment thereof that binds to CDH11 is provided herein, the anti-CDH11 antibody or the fragment thereof that binds to CDH11 comprises a heavy chain variable region and a light chain variable region, the heavy chain and light chain variable regions each comprising CDR1, CDR2, and CDR3, where CDR1H comprises SEQ ID NO:2, CDR2H comprises SEQ ID NO:11, CDR3H comprises SEQ ID NO:22, CDR1L comprises SEQ ID NO:36, CDR2L comprises SEQ ID NO:43, and CDR3L comprises SEQ ID NO:50.

[0039] An anti-CDH11 antibody or a fragment thereof that binds to CDH11 is provided herein, the anti-CDH11 antibody or the fragment thereof that binds to CDH11 comprises a heavy chain variable region and a light chain variable region, the heavy chain and light chain variable regions each comprising CDR1, CDR2, and CDR3, where CDR1H comprises SEQ ID NO:3, CDR2H comprises SEQ ID NO:12, CDR3H comprises SEQ ID NO:23, CDR1L comprises SEQ ID NO:37, CDR2L comprises SEQ ID NO:44, and CDR3L comprises SEQ ID NO:51.

[0040] An anti-CDH11 antibody or a fragment thereof that binds to CDH11 is provided herein, the anti-CDH11 antibody or the fragment thereof that binds to CDH11 comprises a heavy chain variable region and a light chain variable region, the heavy chain and light chain variable regions each comprising CDR1, CDR2, and CDR3, where CDR1H comprises SEQ ID NO:3, CDR2H comprises SEQ ID NO:12, CDR3H comprises SEQ ID NO:24, CDR1L comprises SEQ ID NO:35, CDR2L comprises SEQ ID NO:43, and CDR3L comprises SEQ ID NO:52.

[0041] An anti-CDH11 antibody or a fragment thereof that binds to CDH11 is provided herein, the anti-CDH11 antibody or the fragment thereof that binds to CDH11 comprises a heavy chain variable region and a light chain variable region, the heavy chain and light chain variable regions each comprising CDR1, CDR2, and CDR3, where CDR1H comprises SEQ ID NO: 5, CDR2H comprises SEQ ID NO: 14, CDR3H comprises SEQ ID NO: 27, CDR1L comprises SEQ ID NO: 39, CDR2L comprises SEQ ID NO: 46, and CDR3L comprises SEQ ID NO: 54.

[0042] An anti-CDH11 antibody or a fragment thereof that binds to CDH11 is provided herein, the anti-CDH11 antibody or the fragment thereof that binds to CDH11 comprises a heavy chain variable region and a light chain variable region, the heavy chain and light chain variable regions each comprising CDR1, CDR2, and CDR3, where CDR1H comprises SEQ ID NO: 5, CDR2H comprises SEQ ID NO: 14, CDR3H comprises SEQ ID NO: 28, CDR1L comprises SEQ ID NO: 35, CDR2L comprises SEQ ID NO: 43, and CDR3L comprises SEQ ID NO: 55.

[0043] An anti-CDH11 antibody or a fragment thereof that binds to CDH11 is provided herein, the anti-CDH11 antibody or the fragment thereof that binds to CDH11 comprises a heavy chain variable region and a light chain variable region, the heavy chain and light chain variable regions each comprising CDR1, CDR2, and CDR3, where CDR1H comprises SEQ ID NO: 6, CDR2H comprises SEQ ID NO: 15, CDR3H comprises SEQ ID NO: 29, CDR1L comprises SEQ ID NO: 40, CDR2L comprises SEQ ID NO: 45, and CDR3L comprises SEQ ID NO: 56.

[0044] An anti-CDH11 antibody or a fragment thereof that binds to CDH11 is provided herein, the anti-CDH11 antibody or the fragment thereof that binds to CDH11 comprises a heavy chain variable region and a light chain variable region, the heavy chain and light chain variable regions each comprising CDR1, CDR2, and CDR3, where CDR1H comprises SEQ ID NO:7, CDR2H comprises SEQ ID NO:16, CDR3H comprises SEQ ID NO:30, CDR1L comprises SEQ ID NO:41, CDR2L comprises SEQ ID NO:44, and CDR3L comprises SEQ ID NO:57.

[0045] An anti-CDH11 antibody or a fragment thereof that binds to CDH11 is provided herein, the anti-CDH11 antibody or the fragment thereof that binds to CDH11 comprises a heavy chain variable region and a light chain variable region, the heavy chain and light chain variable regions each comprising CDR1, CDR2, and CDR3, where CDR1H comprises SEQ ID NO:7, CDR2H comprises SEQ ID NO:16, CDR3H comprises SEQ ID NO:31, CDR1L comprises SEQ ID NO:35, CDR2L comprises SEQ ID NO:43, and CDR3L comprises SEQ ID NO:58.

[0046] An anti-CDH11 antibody or a fragment thereof that binds to CDH11 is provided herein, the anti-CDH11 antibody or the fragment thereof that binds to CDH11 comprises a heavy chain variable region and a light chain variable region, the heavy chain and light chain variable regions each comprising CDR1, CDR2, and CDR3, where CDR1H comprises SEQ ID NO:9, CDR2H comprises SEQ ID NO:18, CDR3H comprises SEQ ID NO:33, CDR1L comprises SEQ ID NO:41, CDR2L comprises SEQ ID NO:45, and CDR3L comprises SEQ ID NO:60.

[0047] An anti-CDH11 antibody or a fragment thereof that binds to CDH11 is provided herein, the anti-CDH11 antibody or the fragment thereof that binds to CDH11 comprises a heavy chain variable region and a light chain variable region, the heavy chain and light chain variable regions each comprising CDR1, CDR2, and CDR3, where CDR1H comprises SEQ ID NO:10, CDR2H comprises SEQ ID NO:19, CDR3H comprises SEQ ID NO:34, CDR1L comprises SEQ ID NO:42, CDR2L comprises SEQ ID NO:47, and CDR3L comprises SEQ ID NO:61.

[0048] Anti-CDH11 antibodies or fragments thereof that bind to CDH11 are provided herein and include at least one, at least two, at least three, at least four, at least five or six CDR sequences as disclosed in Tables 1 and 2.

[0049] (Table 1) Summary of the CDR amino acid sequence of the anti-CDH11 antibody. The numbers represent SEQ ID NO. TIFF2026515695000002.tif84146

[0050] (Table 2) Summary of variable chain sequences of anti-CDH11 antibodies. The numbers represent SEQ ID NO. TIFF2026515695000003.tif88146

[0051] In certain embodiments, the intended antibodies and their antigen-binding fragments disclosed herein also feature a humanized framework for reduced immunogenicity. In certain embodiments, the CDR of the intended antibody or its antigen-binding fragment is located within a framework derived from a human antibody or its antigen-binding fragment. In other embodiments, the surface-exposed framework residues of the intended antibody or its antigen-binding fragment are replaced with framework residues of a human antibody or its antigen-binding fragment. The CDR may also be located within a mouse or humanized framework linked to a human constant region (i.e., a chimeric antibody). In one embodiment, the antibody or its antigen-binding fragment comprises a mouse variable region and a human constant region. In one embodiment, the antibody or its antigen-binding fragment comprises a mouse variable region, a mouse CH1 region, and human CH2 and CH3 constant regions. In one embodiment, the CDR of the intended antibody or its antigen-binding fragment is located within a framework that is a complex of two or more human antibodies. In such an embodiment, the intended antibody or its antigen-binding fragment comprises two or more sequence segments ("complexes") derived from the V region of an unrelated human antibody, selected to maintain the monoclonal antibody sequence important for antigen binding of the original precursor monoclonal antibody, and all of these are filtered for potential T cell epitope presentation using "in silico tools" (see, e.g., Holgate & Baker, IDrugs. 2009 Apr;12(4):233-7). The human sequence segments are closely adapted to all sections of the V region of the original antibody, and CD4 is used before synthesis of the antibody or its antigen-binding fragment. + By eliminating T cell epitopes, this technique allows for the avoidance of immunogenicity while maintaining optimal affinity and specificity through pre-analysis of sequences necessary for antigen specificity (Holgate & Baker, 2009).

[0052] Variants of antibody or antigen-binding fragments disclosed herein are provided herein, which share a certain percentage of sequence identity with the antibody or antigen-binding fragments disclosed herein. The term “identity” as used herein refers to sequence identity between two nucleic acid molecules or polypeptides. Identity can be determined by comparing the positions in each sequence that can be aligned for comparison purposes. For example, if the positions in the nucleotide sequences being compared are occupied by the same bases, then the molecules are identical at those positions. The degree of identity between nucleic acid or amino acid sequences is a function of the number of identical or matching nucleotides or amino acids at the shared positions. For example, polypeptides having at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with respect to a particular polypeptide described herein, and preferably exhibiting substantially identical function, as well as polynucleotides encoding such polypeptides. Methods and computer programs for determining both sequence identity and similarity are publicly available and not limited to, the GCG program package (Devereux et al., Nucleic Acids Research 12:387, 1984), BLASTP, BLASTN, FASTA (Altschul et al., J. Mol. Biol. 215:403 (1990), and the ALIGN program (version 2.0). The well-known Smith-Waterman algorithm may also be used to determine similarity. The BLAST program is publicly available from NCBI and other sources (BLAST Manual, Altschul, et al., NCBI NLM NIH, Bethesda, Md. 20894; BLAST 2.0 at http: / / www.ncbi.nlm.nih.gov / blast / ). When comparing sequences, these methods will account for various substitutions, deletions, and other modifications.

[0053] In some embodiments, an anti-CDH11 antibody or a fragment thereof that binds to CDH11 is provided, comprising a heavy chain variable domain containing a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to any one of SEQ ID NO: 62-77.

[0054] In some embodiments, an anti-CDH11 antibody or a fragment thereof that binds to CDH11 is provided, comprising a light chain variable domain containing a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to any one of SEQ ID NO: 78-93.

[0055] In some embodiments, an anti-CDH11 antibody or a fragment thereof that binds to CDH11 is provided. (a) A heavy chain variable domain containing a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to any one of SEQ ID NO: 62-77, and (b) A light chain variable domain containing a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to any one of SEQ ID NO: 78-93.

[0056] An anti-CDH11 antibody or a fragment thereof that binds to CDH11 is provided herein, comprising a heavy chain variable domain containing a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO:62, and a light chain variable domain containing a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO:78.

[0057] An anti-CDH11 antibody or a fragment thereof that binds to CDH11 is provided herein, comprising a heavy chain variable domain containing a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO:68, and a light chain variable domain containing a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO:83.

[0058] An anti-CDH11 antibody or a fragment thereof that binds to CDH11 is provided herein, comprising a heavy chain variable domain containing a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO:75, and a light chain variable domain containing a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO:90.

[0059] An anti-CDH11 antibody or a fragment thereof that binds to CDH11 is provided herein, comprising a heavy chain variable domain containing a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO:63, and a light chain variable domain containing a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO:79.

[0060] An anti-CDH11 antibody or a fragment thereof that binds to CDH11 is provided herein, comprising a heavy chain variable domain containing a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO:64, and a light chain variable domain containing a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO:80.

[0061] An anti-CDH11 antibody or a fragment thereof that binds to CDH11 is provided herein, comprising a heavy chain variable domain containing a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO:65, and a light chain variable domain containing a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO:81.

[0062] An anti-CDH11 antibody or a fragment thereof that binds to CDH11 is provided herein, comprising a heavy chain variable domain containing a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO:66, and a light chain variable domain containing a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO:82.

[0063] An anti-CDH11 antibody or a fragment thereof that binds to CDH11 is provided herein, comprising a heavy chain variable domain containing a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO:67, and a light chain variable domain containing a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO:83.

[0064] An anti-CDH11 antibody or a fragment thereof that binds to CDH11 is provided herein, comprising a heavy chain variable domain containing a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO:69, and a light chain variable domain containing a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO:84.

[0065] An anti-CDH11 antibody or a fragment thereof that binds to CDH11 is provided herein, comprising a heavy chain variable domain containing a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO:70, and a light chain variable domain containing a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO:85.

[0066] An anti-CDH11 antibody or a fragment thereof that binds to CDH11 is provided herein, comprising a heavy chain variable domain containing a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO:71, and a light chain variable domain containing a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO:86.

[0067] An anti-CDH11 antibody or a fragment thereof that binds to CDH11 is provided herein, comprising a heavy chain variable domain containing a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO:72, and a light chain variable domain containing a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO:87.

[0068] An anti-CDH11 antibody or a fragment thereof that binds to CDH11 is provided herein, comprising a heavy chain variable domain containing a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO:73, and a light chain variable domain containing a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO:88.

[0069] An anti-CDH11 antibody or a fragment thereof that binds to CDH11 is provided herein, comprising a heavy chain variable domain containing a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO:74, and a light chain variable domain containing a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO:89.

[0070] An anti-CDH11 antibody or a fragment thereof that binds to CDH11 is provided herein, comprising a heavy chain variable domain containing a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO:76, and a light chain variable domain containing a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO:91.

[0071] An anti-CDH11 antibody or a fragment thereof that binds to CDH11 is provided herein, comprising a heavy chain variable domain containing a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO:76, and a light chain variable domain containing a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO:92.

[0072] An anti-CDH11 antibody or a fragment thereof that binds to CDH11 is provided herein, comprising a heavy chain variable domain containing a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO:77, and a light chain variable domain containing a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO:93.

[0073] An anti-CDH11 antibody or a fragment thereof that binds to CDH11 is provided herein, comprising a heavy chain variable region including SEQ ID NO:62 and a light chain variable region including SEQ ID NO:78.

[0074] An anti-CDH11 antibody or a fragment thereof that binds to CDH11 is provided herein, comprising a heavy chain variable region including SEQ ID NO:68 and a light chain variable region including SEQ ID NO:83.

[0075] An anti-CDH11 antibody or a fragment thereof that binds to CDH11 is provided herein, comprising a heavy chain variable region including SEQ ID NO:75 and a light chain variable region including SEQ ID NO:90.

[0076] An anti-CDH11 antibody or a fragment thereof that binds to CDH11 is provided herein, comprising a heavy chain variable region including SEQ ID NO:63 and a light chain variable region including SEQ ID NO:79.

[0077] An anti-CDH11 antibody or a fragment thereof that binds to CDH11 is provided herein, comprising a heavy chain variable region including SEQ ID NO:64 and a light chain variable region including SEQ ID NO:80.

[0078] An anti-CDH11 antibody or a fragment thereof that binds to CDH11 is provided herein, comprising a heavy chain variable region including SEQ ID NO:65 and a light chain variable region including SEQ ID NO:81.

[0079] An anti-CDH11 antibody or a fragment thereof that binds to CDH11 is provided herein, comprising a heavy chain variable region including SEQ ID NO:66 and a light chain variable region including SEQ ID NO:82.

[0080] An anti-CDH11 antibody or a fragment thereof that binds to CDH11 is provided herein, comprising a heavy chain variable region including SEQ ID NO:67 and a light chain variable region including SEQ ID NO:83.

[0081] An anti-CDH11 antibody or a fragment thereof that binds to CDH11 is provided herein, comprising a heavy chain variable region including SEQ ID NO:69 and a light chain variable region including SEQ ID NO:84.

[0082] An anti-CDH11 antibody or a fragment thereof that binds to CDH11 is provided herein, comprising a heavy chain variable region including SEQ ID NO:70 and a light chain variable region including SEQ ID NO:85.

[0083] An anti-CDH11 antibody or a fragment thereof that binds to CDH11 is provided herein, comprising a heavy chain variable region including SEQ ID NO:71 and a light chain variable region including SEQ ID NO:86.

[0084] An anti-CDH11 antibody or a fragment thereof that binds to CDH11 is provided herein, comprising a heavy chain variable region including SEQ ID NO:72 and a light chain variable region including SEQ ID NO:87.

[0085] An anti-CDH11 antibody or a fragment thereof that binds to CDH11 is provided herein, comprising a heavy chain variable region including SEQ ID NO:73 and a light chain variable region including SEQ ID NO:88.

[0086] An anti-CDH11 antibody or a fragment thereof that binds to CDH11 is provided herein, comprising a heavy chain variable region including SEQ ID NO:74 and a light chain variable region including SEQ ID NO:89.

[0087] An anti-CDH11 antibody or a fragment thereof that binds to CDH11 is provided herein, comprising a heavy chain variable region including SEQ ID NO:76 and a light chain variable region including SEQ ID NO:91.

[0088] An anti-CDH11 antibody or a fragment thereof that binds to CDH11 is provided herein, comprising a heavy chain variable region including SEQ ID NO:76 and a light chain variable region including SEQ ID NO:92.

[0089] An anti-CDH11 antibody or a fragment thereof that binds to CDH11 is provided herein, comprising a heavy chain variable region including SEQ ID NO:77 and a light chain variable region including SEQ ID NO:93.

[0090] An anti-CDH11 antibody or a fragment thereof that binds to CDH11 is provided herein, and the anti-CDH11 antibody or a fragment thereof that binds to CDH11 comprises a heavy chain variable region and a light chain variable region, each of the heavy chain and light chain variable regions comprising CDR1, CDR2, and CDR3, where (a) CDR1H comprises SEQ ID NO:1, CDR2H comprises SEQ ID NO:11, CDR3H comprises SEQ ID NO:20, CDR1L comprises SEQ ID NO:35, CDR2L comprises SEQ ID NO:43, and CDR3L comprises SEQ ID NO:48, (b) the heavy chain variable region comprises a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO:62, and (c) the light chain variable region comprises SEQ ID Contains sequences that are at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to NO:78.

[0091] An anti-CDH11 antibody or a fragment thereof that binds to CDH11 is provided herein, and the anti-CDH11 antibody or a fragment thereof that binds to CDH11 comprises a heavy chain variable region and a light chain variable region, each of the heavy chain and light chain variable regions comprising CDR1, CDR2, and CDR3, (a) CDR1H comprises SEQ ID NO:1, CDR2H comprises SEQ ID NO:11, CDR3H comprises SEQ ID NO:21, CDR1L comprises SEQ ID NO:35, CDR2L comprises SEQ ID NO:43, and CDR3L comprises SEQ ID NO:49, (b) the heavy chain variable region comprises a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO:63, and (c) the light chain variable region comprises SEQ ID Contains sequences that are at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to NO:79.

[0092] An anti-CDH11 antibody or a fragment thereof that binds to CDH11 is provided herein, and the anti-CDH11 antibody or a fragment thereof that binds to CDH11 comprises a heavy chain variable region and a light chain variable region, each of the heavy chain and light chain variable regions comprising CDR1, CDR2, and CDR3, (a) CDR1H comprises SEQ ID NO:4, CDR2H comprises SEQ ID NO:13, CDR3H comprises SEQ ID NO:25, CDR1L comprises SEQ ID NO:38, CDR2L comprises SEQ ID NO:45, and CDR3L comprises SEQ ID NO:53, (b) the heavy chain variable region comprises a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO:67, and (c) the light chain variable region comprises SEQ ID Contains sequences that are at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to NO:83.

[0093] An anti-CDH11 antibody or a fragment thereof that binds to CDH11 is provided herein, and the anti-CDH11 antibody or a fragment thereof that binds to CDH11 comprises a heavy chain variable region and a light chain variable region, each of the heavy chain and light chain variable regions comprising CDR1, CDR2, and CDR3, (a) CDR1H comprises SEQ ID NO:4, CDR2H comprises SEQ ID NO:13, CDR3H comprises SEQ ID NO:26, CDR1L comprises SEQ ID NO:38, CDR2L comprises SEQ ID NO:45, and CDR3L comprises SEQ ID NO:53, (b) the heavy chain variable region comprises a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO:68, and (c) the light chain variable region comprises SEQ ID Contains sequences that are at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to NO:83.

[0094] An anti-CDH11 antibody or a fragment thereof that binds to CDH11 is provided herein, and the anti-CDH11 antibody or a fragment thereof that binds to CDH11 comprises a heavy chain variable region and a light chain variable region, each of the heavy chain and light chain variable regions comprising CDR1, CDR2, and CDR3, (a) CDR1H comprises SEQ ID NO: 8, CDR2H comprises SEQ ID NO: 17, CDR3H comprises SEQ ID NO: 32, CDR1L comprises SEQ ID NO: 38, CDR2L comprises SEQ ID NO: 45, and CDR3L comprises SEQ ID NO: 59, (b) the heavy chain variable region comprises a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 75, and (c) the light chain variable region comprises SEQ ID Contains sequences that are at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to NO:90.

[0095] An anti-CDH11 antibody or a fragment thereof that binds to CDH11 is provided herein, and the anti-CDH11 antibody or a fragment thereof that binds to CDH11 comprises a heavy chain variable region and a light chain variable region, each of the heavy chain and light chain variable regions comprising CDR1, CDR2, and CDR3, (a) CDR1H comprises SEQ ID NO: 8, CDR2H comprises SEQ ID NO: 17, CDR3H comprises SEQ ID NO: 32, CDR1L comprises SEQ ID NO: 38, CDR2L comprises SEQ ID NO: 45, and CDR3L comprises SEQ ID NO: 59, (b) the heavy chain variable region comprises a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 74, and (c) the light chain variable region comprises SEQ ID Contains sequences that are at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to NO:89.

[0096] An anti-CDH11 antibody or a fragment thereof that binds to CDH11 is provided herein, and the anti-CDH11 antibody or a fragment thereof that binds to CDH11 comprises a heavy chain variable region and a light chain variable region, each of the heavy chain and light chain variable regions comprising CDR1, CDR2, and CDR3, (a) CDR1H comprises SEQ ID NO:2, CDR2H comprises SEQ ID NO:11, CDR3H comprises SEQ ID NO:22, CDR1L comprises SEQ ID NO:36, CDR2L comprises SEQ ID NO:43, and CDR3L comprises SEQ ID NO:50, (b) the heavy chain variable region comprises a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO:64, and (c) the light chain variable region comprises SEQ ID Contains sequences that are at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to NO:80.

[0097] An anti-CDH11 antibody or a fragment thereof that binds to CDH11 is provided herein, and the anti-CDH11 antibody or a fragment thereof that binds to CDH11 comprises a heavy chain variable region and a light chain variable region, each of the heavy chain and light chain variable regions comprising CDR1, CDR2, and CDR3, (a) CDR1H comprises SEQ ID NO:3, CDR2H comprises SEQ ID NO:12, CDR3H comprises SEQ ID NO:23, CDR1L comprises SEQ ID NO:37, CDR2L comprises SEQ ID NO:44, and CDR3L comprises SEQ ID NO:51, (b) the heavy chain variable region comprises a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO:65, and (c) the light chain variable region comprises SEQ ID Contains sequences that are at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to NO:81.

[0098] An anti-CDH11 antibody or a fragment thereof that binds to CDH11 is provided herein, and the anti-CDH11 antibody or a fragment thereof that binds to CDH11 comprises a heavy chain variable region and a light chain variable region, each of the heavy chain and light chain variable regions comprising CDR1, CDR2, and CDR3, (a) CDR1H comprises SEQ ID NO:3, CDR2H comprises SEQ ID NO:12, CDR3H comprises SEQ ID NO:24, CDR1L comprises SEQ ID NO:35, CDR2L comprises SEQ ID NO:43, and CDR3L comprises SEQ ID NO:52, (b) the heavy chain variable region comprises a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO:66, and (c) the light chain variable region comprises SEQ ID Contains sequences that are at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to NO:82.

[0099] An anti-CDH11 antibody or a fragment thereof that binds to CDH11 is provided herein, and the anti-CDH11 antibody or a fragment thereof that binds to CDH11 comprises a heavy chain variable region and a light chain variable region, each of the heavy chain and light chain variable regions comprising CDR1, CDR2, and CDR3, (a) CDR1H comprises SEQ ID NO: 5, CDR2H comprises SEQ ID NO: 14, CDR3H comprises SEQ ID NO: 27, CDR1L comprises SEQ ID NO: 39, CDR2L comprises SEQ ID NO: 46, and CDR3L comprises SEQ ID NO: 54, (b) the heavy chain variable region comprises a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 69, and (c) the light chain variable region comprises SEQ ID Contains sequences that are at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to NO:84.

[0100] An anti-CDH11 antibody or a fragment thereof that binds to CDH11 is provided herein, and the anti-CDH11 antibody or a fragment thereof that binds to CDH11 comprises a heavy chain variable region and a light chain variable region, each of the heavy chain and light chain variable regions comprising CDR1, CDR2, and CDR3, (a) CDR1H comprises SEQ ID NO: 5, CDR2H comprises SEQ ID NO: 14, CDR3H comprises SEQ ID NO: 28, CDR1L comprises SEQ ID NO: 35, CDR2L comprises SEQ ID NO: 43, and CDR3L comprises SEQ ID NO: 55, (b) the heavy chain variable region comprises a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 70, and (c) the light chain variable region comprises SEQ ID Contains sequences that are at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to NO:85.

[0101] An anti-CDH11 antibody or a fragment thereof that binds to CDH11 is provided herein, and the anti-CDH11 antibody or a fragment thereof that binds to CDH11 comprises a heavy chain variable region and a light chain variable region, each of the heavy chain and light chain variable regions comprising CDR1, CDR2, and CDR3, (a) CDR1H comprises SEQ ID NO: 6, CDR2H comprises SEQ ID NO: 15, CDR3H comprises SEQ ID NO: 29, CDR1L comprises SEQ ID NO: 40, CDR2L comprises SEQ ID NO: 45, and CDR3L comprises SEQ ID NO: 56, (b) the heavy chain variable region comprises a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 71, and (c) the light chain variable region comprises SEQ ID Contains sequences that are at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to NO:86.

[0102] An anti-CDH11 antibody or a fragment thereof that binds to CDH11 is provided herein, and the anti-CDH11 antibody or a fragment thereof that binds to CDH11 comprises a heavy chain variable region and a light chain variable region, each of the heavy chain and light chain variable regions comprising CDR1, CDR2, and CDR3, (a) CDR1H comprises SEQ ID NO:7, CDR2H comprises SEQ ID NO:16, CDR3H comprises SEQ ID NO:30, CDR1L comprises SEQ ID NO:41, CDR2L comprises SEQ ID NO:44, and CDR3L comprises SEQ ID NO:57, (b) the heavy chain variable region comprises a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO:72, and (c) the light chain variable region comprises SEQ ID Contains sequences that are at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to NO:87.

[0103] An anti-CDH11 antibody or a fragment thereof that binds to CDH11 is provided herein, and the anti-CDH11 antibody or a fragment thereof that binds to CDH11 comprises a heavy chain variable region and a light chain variable region, each of the heavy chain and light chain variable regions comprising CDR1, CDR2, and CDR3, (a) CDR1H comprises SEQ ID NO:7, CDR2H comprises SEQ ID NO:16, CDR3H comprises SEQ ID NO:31, CDR1L comprises SEQ ID NO:35, CDR2L comprises SEQ ID NO:43, and CDR3L comprises SEQ ID NO:58, (b) the heavy chain variable region comprises a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO:73, and (c) the light chain variable region comprises SEQ ID Contains sequences that are at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to NO:88.

[0104] An anti-CDH11 antibody or a fragment thereof that binds to CDH11 is provided herein, and the anti-CDH11 antibody or a fragment thereof that binds to CDH11 comprises a heavy chain variable region and a light chain variable region, each of the heavy chain and light chain variable regions comprising CDR1, CDR2, and CDR3, (a) CDR1H comprises SEQ ID NO:9, CDR2H comprises SEQ ID NO:18, CDR3H comprises SEQ ID NO:33, CDR1L comprises SEQ ID NO:41, CDR2L comprises SEQ ID NO:45, and CDR3L comprises SEQ ID NO:60, (b) the heavy chain variable region comprises a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO:76, and (c) the light chain variable region comprises SEQ ID Contains sequences that are at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to NO:91.

[0105] An anti-CDH11 antibody or a fragment thereof that binds to CDH11 is provided herein, and the anti-CDH11 antibody or a fragment thereof that binds to CDH11 comprises a heavy chain variable region and a light chain variable region, each of the heavy chain and light chain variable regions comprising CDR1, CDR2, and CDR3, (a) CDR1H comprises SEQ ID NO:9, CDR2H comprises SEQ ID NO:18, CDR3H comprises SEQ ID NO:33, CDR1L comprises SEQ ID NO:41, CDR2L comprises SEQ ID NO:45, and CDR3L comprises SEQ ID NO:60, (b) the heavy chain variable region comprises a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO:76, and (c) the light chain variable region comprises SEQ ID Contains sequences that are at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to NO:92.

[0106] An anti-CDH11 antibody or a fragment thereof that binds to CDH11 is provided herein, and the anti-CDH11 antibody or a fragment thereof that binds to CDH11 comprises a heavy chain variable region and a light chain variable region, each of the heavy chain and light chain variable regions comprising CDR1, CDR2, and CDR3, (a) CDR1H comprises SEQ ID NO:10, CDR2H comprises SEQ ID NO:19, CDR3H comprises SEQ ID NO:34, CDR1L comprises SEQ ID NO:42, CDR2L comprises SEQ ID NO:47, and CDR3L comprises SEQ ID NO:61, (b) the heavy chain variable region comprises a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO:77, and (c) the light chain variable region comprises SEQ ID Contains sequences that are at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to NO:93.

[0107] Antibodies disclosed herein may include modifications in their variable and constant regions. For example, antibodies containing a modified but functionally equivalent variable region and / or CDR are provided herein. In some embodiments, the modifications do not significantly affect the properties of the antibody or its antigen-binding fragment. In some embodiments, the modifications result in enhanced or reduced activity and / or affinity. For example, the amino acid sequence of an anti-CDH11 antibody or its fragment that binds to CDH11 may be mutated to obtain an antibody having a desired binding affinity to CDH11. Modification of polypeptides is a common practice to those skilled in the art and does not need to be described in detail herein. Examples of modified polypeptides include polypeptides having conservative substitutions of amino acid residues, polypeptides having deletions or additions of one or more amino acids that do not significantly or dramatically alter functional activity or mature (enhance) the polypeptide's affinity to its ligand, or polypeptides having the use of chemical analogs.

[0108] Amino acid sequence insertions include fusions at the amino and / or carboxyl terminals of polypeptides ranging in length from one residue to 100 or more residues, as well as intrasequences of single or multiple amino acid residues. Examples of terminal insertions include antibodies with an N-terminal methionyl residue or antibodies fused to an epitope tag. Other insertion variants of antibody molecules include fusion of the antibody or its antigen-binding fragment to the N-terminus or C-terminus by an enzyme or polypeptide that increases the antibody's half-life in the bloodstream.

[0109] Substitutional variants of antibodies and their antigen-binding fragments have at least one amino acid residue removed from the antibody molecule and a different residue introduced in its place. Substitutional mutagenesis may be directed to the hypervariable region, but framework changes are also intended. Examples of conservative substitutions are shown in Table 3.

[0110] (Table 3) Conservative amino acid substitutions TIFF2026515695000004.tif98146

[0111] Naturally occurring residues can be divided into several groups based on common side-chain characteristics: (1) Nonpolar: norleucine, Met, Ala, Val, Leu, Ile; (2) Polar, uncharged: Cys, Ser, Thr, Asn, Gln; (3) Acidic (negative charge): Asp, Glu; (4) Basic (positive charge): Lys, Arg; (5) Residues that affect chain orientation: Gly, Pro; and (6) Aromatic: Trp, Tyr, Phe, His.

[0112] Conservative substitutions can also be created by exchanging one member of one of these classes with another member of that class.

[0113] One type of substitution that can be made is, for example, to replace one or more cysteines in an antibody, which may be chemically reactive, with another residue, such as alanine or serine, but not limited to the following. For example, it may be a substitution of a non-canonical cysteine. The substitution can be made in the CDR or framework region of the variable domain of the antibody, or in the constant region. In some embodiments, the cysteine ​​is canonical. Any cysteine ​​residue that does not participate in maintaining the proper three-dimensional structure of the antibody may also be substituted with serine, generally to improve the oxidative stability of the molecule and prevent abnormal crosslinking. Conversely, cysteine ​​bonds may be added to the antibody to improve its stability, especially when the antibody is an antibody fragment, such as an Fv fragment.

[0114] In several embodiments, the antibodies of this disclosure are monoclonal antibodies or antigen-binding fragments thereof, or fragments of monoclonal antibodies. As used herein, the term “monoclonal antibody” refers to an antibody member that is a substantially homogeneous population of antibodies, i.e., the individual antibodies constituting the population are identical except for possible mutations, e.g., naturally occurring mutations that may be present in trace amounts. Thus, the modifying phrase “monoclonal” indicates that the antibody features are not a mixture of distinct antibodies. In certain embodiments, such a monoclonal antibody typically comprises an antibody containing a polypeptide sequence that binds to a target on CDH11, and the target-binding polypeptide sequence is obtained by a process that includes the selection of a single target-binding polypeptide sequence from a plurality of polypeptide sequences. For example, the selection process may be the selection of a specific clone from a pool of multiple clones, e.g., hybridoma clones, phage clones, or recombinant DNA clones. In contrast to polyclonal antibody preparations, which typically contain different antibodies that are directed to different determinants (epitopes), each monoclonal antibody in a monoclonal antibody preparation is directed to a single determinant of a given antigen. In addition to their specificity, monoclonal antibody preparations have the advantage of not being contaminated with other immunoglobulins. Therefore, the specified monoclonal antibody can be produced by non-hybridoma techniques, for example, by appropriate recombination methods once its sequence has been identified.

[0115] In some embodiments, the antibodies disclosed herein are human antibodies and / or their antigen-binding fragments, or fragments of human antibodies. As used herein, “human antibody” corresponds to (i.e., is sequence-identical to) antibodies that can be produced by humans and / or antibodies produced using any techniques for producing human antibodies disclosed herein, but which are not produced in humans.The "human antibodies" used herein are obtained by the method described in Hoogenboom and Winter, J. Mol. Biol., 227:381 (1991); Marks et al., J. Mol. Biol., 222:581 (1991), which is incorporated herein by reference in its entirety; Cole et al., Monoclonal Antibodies and Cancer Therapy, Alan R. Liss, p. 77 (1985), which is incorporated herein by reference in its entirety; Boerner et al., J. Immunol., 147(1):86-95 (1991), which is incorporated herein by reference in its entirety; and van Dijk and van de Winkel, Curr. Opin. Pharmacol., which is incorporated herein by reference in its entirety. Using various techniques known to those skilled in the art, including 5:368-74 (2001), and an antigen (e.g., CDH11 or entities containing it), genetically modified animals, such as immunized xenograft mice (see, for example, U.S. Patent Nos. 5,939,598; 6,075,181; 6,114,598; 6,150,584 and 6,162,963, each of which is incorporated herein by reference in whole, each of the following patents: VelocImmune® (Regeneron, Tarrytown, NY), UltiMab® platform (Medarex, now Bristol Myers) It can be produced by administering it to Squibb (Princeton, NJ). See also, for example, Li et al., Proc. Natl. Acad. Sci. USA, 103:3557-3562 (2006) regarding human antibodies produced via human B-cell hybridoma technology.See also the KM Mouse® system described in PCT Publication WO 02 / 43478 by Ishida et al., in which the mouse possesses a human heavy chain transchromosome and a human light chain transgene, both of which are incorporated herein by reference in their entirety, and the TC mouse system described in Tomizuka et al. (2000) Proc. Natl. Acad. Sci. USA 97:722-727, in which the mouse possesses both a human heavy chain transchromosome and a human light chain transchromosome. In each of these systems, the transgene and / or transchromosome possessed by the mouse contains sequences of the variable and constant regions of human immunoglobulins. As used herein, the term “human antibody” is intended to include antibodies that have a variable region, in which both the framework and CDR region are sequences of human origin or identical sequences, but are not naturally occurring in humans or produced in humans. Furthermore, if the antibody (e.g., an intact antibody other than a Fab fragment) contains a constant region, this constant region is derived from such a human sequence, e.g., a human germline sequence, or a mutated version of a human germline sequence. Human antibodies disclosed herein may contain amino acid residues not encoded by a human sequence (e.g., mutations introduced in vitro by random or site-directed mutagenesis, or in vivo by somatic mutation). In one non-limiting embodiment, the human antibody is a human monoclonal antibody, and such an antibody can be produced by a hybridoma fused with immortalized cells containing B cells obtained from a transgenic non-human animal, e.g., a transgenic mouse having a genome containing human heavy chain and light chain transgenes.

[0116] In several embodiments, the antibody or its antigen-binding fragment is a recombinant antibody. As used herein, the term “recombinant human antibody” includes all human antibodies prepared, expressed, produced, or isolated by recombinant means, such as antibodies isolated from animals (e.g., mice) that have been genetically modified or transfected for human immunoglobulin genes, or antibodies isolated from hybridomas prepared therefrom, antibodies isolated from host cells transformed to express human antibodies, such as transfectomas, antibodies isolated from recombinant combinatorial human antibody libraries, and antibodies prepared, expressed, produced, or isolated by any other means, including splicing all or part of human immunoglobulin gene sequences to other DNA sequences. Such recombinant human antibodies have a variable region in which the framework and CDR region are derived from human germline immunoglobulin sequences. However, in certain embodiments, such recombinant human antibodies may be subject to in vitro mutagenesis (or in vivo somatic mutagenesis if an animal transgene is used for the human Ig sequence), and therefore the amino acid sequences of the VH and VL regions of the recombinant antibody may be derived from and related to the VH and VL sequences of the human germline, but which may not naturally exist in the repertoire of human antibody germlines in vivo.

[0117] In some embodiments, the antibody or its antigen-binding fragment is isolated. As used herein, “isolated antibody” means an antibody that, depending on its origin or source, satisfies one, two, three, or four of the following criteria: (1) substantially unrelated to any naturally occurring components in its natural state, (2) substantially free from other proteins from the same species, (3) expressed by cells from a different species, and (4) not present in nature.

[0118] In several embodiments, the antibody or its antigen-binding fragment includes a human constant region or a modified constant region. In some embodiments, the antibody or antigen-binding fragment has a non-human constant region or a modified non-human constant region. In one embodiment, the antibody or its antigen-binding fragment has a mouse constant region or a modified mouse constant region. In one embodiment, the antibody or its antigen-binding fragment has a non-human primate constant region or a modified non-human primate constant region. In some embodiments, the antibody or its antigen-binding fragment has a non-human constant region or a modified non-human constant region. In some embodiments, the constant region is derived from a non-human primate, mouse, rat, sheep, goat, or rabbit.

[0119] Depending on the amino acid sequence of the constant domain of its heavy chain, antibodies (immunoglobulins) can be assigned to different classes. Each antibody or its antigen-binding fragment may be, for example, an IgG, IgD, IgE, IgA, or IgM antibody or a fragment thereof. In one embodiment, the antibody is immunoglobulin G. In one embodiment, the antibody fragment is a fragment of immunoglobulin G. In one embodiment, the antibody is IgG1, IgG2, IgG2a, IgG2b, IgG3, or IgG4. In one embodiment, the antibody contains a sequence from mammalian IgG1, mammalian IgG2, mammalian IgG3, or mammalian IgG4. In one embodiment, the antibody contains a sequence from human IgG1, human IgG2, human IgG3, or human IgG4. Any combination of these antibody subtypes can also be used. One consideration in selecting the type of antibody to be used is the desired serum half-life of the antibody. For example, IgG generally has a serum half-life of 23 days, IgA 6 days, IgM 5 days, IgD 3 days, and IgE 2 days (the whole is incorporated herein by reference, Abbas AK, Lichtman AH, Pober JS. Cellular and Molecular Immunology, 4th edition, WB Saunders Co., Philadelphia, 2000). Antibodies or their antigen-binding fragments may contain sequences from one or more classes or isotypes.

[0120] In one embodiment, the antibody or its antigen-binding fragment contains an Fc domain with the same sequence as the human IgG1 Fc domain or having 99% or more sequence similarity thereto. In one embodiment, the antibody or its antigen-binding fragment contains an Fc domain with the same sequence as the human IgG2 Fc domain or having 99% or more sequence similarity thereto. In one embodiment, the antibody or its antigen-binding fragment contains an Fc domain with the same sequence as the human IgG3 Fc domain or having 99% or more sequence similarity thereto. In one embodiment, the antibody or its antigen-binding fragment contains an Fc domain with the same sequence as the human IgG4 Fc domain or having 99% or more sequence similarity thereto. In one embodiment, the Fc domain is unmutated.

[0121] In one embodiment, the Fc domain is mutated at the interface of the CH2-CH3 domain such that the affinity of IgG for FcRn increases in acidic conditions, but not at neutral pH. In another embodiment, the antibody or its antigen-binding fragment contains an Fc domain having the same sequence as the human IgG1 Fc domain.

[0122] In some embodiments, the antibody or its antigen-binding fragment has a modified IgG Fc region. Modified IgG Fc regions are well known in the art. See, for example, any of the mutations listed in Table 1 of Wang et al. Protein Cell (2018), 9(1):63-73. In several embodiments, compared to an unmodified Fc region, the modified Fc region has enhanced complement-based effector function, increased or decreased FcγR-based effector function, reduced effector function, enhanced co-association with antigen and FcγR, and / or increased serum half-life.

[0123] Non-limiting examples of Fc modifications to alter the effector function of antibodies against IgG1 (see Wang et al. Protein Cell (2018), 9(1):63-73) include: (a) Increased FcγRIIIa binding: F243L / R292P / Y300L / V305I / P396L (b) Increased FcγRIIIa binding: S239D / I332E (c) Increased FcγRIIIa binding: Decreased FcγRIIb binding, S239D / I332E / A330L (d) Increased FcγRIIIa binding: S298A / E333A / K334A i. For example, in one heavy chain: L234Y / L235Q / G236W / S239M / H268D / D270E / S298A ii. For example, in opposing heavy chains: D270E / K326D / A330M / K334E (e) Increased FcγRIIa binding, increased FcγRIIIa binding: G236A / S239D / I332E (f) Enhanced CDC, increased C1q binding: K326W / E333S (g) Increased C1q bond: S267E / H268F / S324T (h) Increased C1q binding, IgG1 / IgG3 cross-subclass hexamerization: E345R / E430G / S440Y (i) Reduced effector function: Deglycosylation N297A or N297Q or N297G (j) Reduced FcγR and C1q binding: L235E (k) Reduced FcγR and C1q binding i. IgG1:L234A / L235A ii. IgG4:F234A / L235A (l) Reduced FcγR and C1q binding, IgG2 / IgG4 cross-isotype (m) Reduced FcγR and C1q-bound IgG2:H268Q / V309L / A330S / P331S (n) Reduced FcγR and C1q-bound IgG2: V234A / G237A / P238S / H268A / V309L / A330S / P331S (o) Increased FcRn binding at pH 6.0: M252Y / S254T / T256E (p) Increased FcRn binding at pH 6.0: M428L / N434S (q) Increased FcγRIIb binding: S267E / L328F (r) Increased FcγRIIa binding, decreased FcγRIIIa binding: N325S / L328F.

[0124] Antibody binding Anti-CDH11 antibodies or antigen-binding fragments thereof that bind to human CDH11 are provided herein. Anti-CDH11 antibodies or antigen-binding fragments thereof that bind to mouse CDH11 are provided herein. Anti-CDH11 antibodies or antigen-binding fragments thereof that bind to both human CDH11 and mouse CDH11 are provided herein. Anti-CDH11 antibodies or antigen-binding fragments thereof that bind to the extracellular domain 3 (extracellular cadherin domain, EC3) of CDH11 are provided herein. Anti-CDH11 antibodies or antigen-binding fragments thereof that bind to EC4 of CDH11 are provided herein. Anti-CDH11 antibodies or antigen-binding fragments thereof that bind to EC5 of CDH11 are provided herein. Anti-CDH11 antibodies or antigen-binding fragments thereof that bind to EC3 of human CDH11 are provided herein. Anti-CDH11 antibodies or antigen-binding fragments thereof that bind to EC4 of human CDH11 are provided herein. Anti-CDH11 antibodies or antigen-binding fragments thereof that bind to EC5 of human CDH11 are provided herein. Anti-CDH11 antibodies or antigen-binding fragments thereof that bind to EC3 of mouse CDH11 are provided herein. Anti-CDH11 antibodies or antigen-binding fragments thereof that bind to EC4 of mouse CDH11 are provided herein. Anti-CDH11 antibodies or antigen-binding fragments thereof that bind to EC5 of mouse CDH11 are provided herein.

[0125] In multiple embodiments, the anti-CDH11 antibody or fragment thereof that binds to CDH11 is from about 1x10 -6 M to about 1x10 -10 M, about 1x10 -7 M to about 1x10 -10 M, about 1x10-8 M ~ approx. 1x10 -10 M, about 1x10- 8 M ~ approx. 1x10 -19 Binding affinity of M (K D ) binds to CDH11. In multiple embodiments, the fragment that binds to the anti-CDH11 antibody or CDH11 is approximately 1 x 10⁻¹⁰. -7 M, about 1x10- 8 M, about 1x10 -9 M, or approximately 1x10 -10 Binding affinity of M (K D ) binds to CDH11. In one embodiment, the anti-CDH11 antibody or the fragment thereof that binds to CDH11 as described herein binds to CDH11 with a dissociation constant of <1 μM, <1 nM, or <10 pM. In one embodiment, the anti-CDH11 antibody or the fragment thereof that binds to CDH11 has a K for CDH11 D It is less than 100 nM. In one embodiment, the K of the anti-CDH11 antibody or its fragment that binds to CDH11, against CDH11 D It is less than 10 nM. In one embodiment, the K of the anti-CDH11 antibody or its fragment that binds to CDH11, against CDH11 D It is less than 1.0 nM. The term "K" as used herein D " is intended to refer to the dissociation constant of the antibody-antigen interaction. The K of the antibody against that antigen D Alternatively, one way to determine binding affinity is by measuring the binding affinity using a Dip and Read assay (Octet Red96 ForteBio, Fremont, CA) with immobilized and monoclonal antibodies (the affinity constant is the reciprocal of the dissociation constant). Biotinylated antibodies can be diluted in PBS + 0.1% BSA, 0.02% Tween20, and 0.05% sodium azide (Kinetics Buffer, ForteBio) and immersed in wells containing serially diluted mAbs, starting at 37.75 nM. The concentration of Fab protein is determined by ELISA and / or SDS-PAGE electrophoresis using a known concentration of IgG1 standard monoclonal antibody as a standard. The kinetic association rate (kon ) and dissociation rate (k off The equilibrium dissociation constant (K) is obtained simultaneously by using the BIA evaluation program and fitting the data to a 1:1 Langmuir coupled model (the entire contents of which are incorporated herein by reference, Karlsson, R. Roos, H. Fagerstam, L. Petersson, B. (1994). Methods Enzymology 6. 99-110). D The value of ) is k off / k on It is calculated as follows. This protocol is suitable for use in determining the binding affinity of an antibody or fragment to any antigen. Other protocols known in the art may also be used. For example, K D To determine the value, an ELISA for antigens using mAbs can be used.

[0126] The term "specifically binding" to an antibody or polypeptide is a well-understood term in the art. A molecular entity is said to exhibit "specific binding" or "preferred binding" if it reacts with or associates with a particular cell or substance more frequently, more quickly, for a longer period, and / or with higher affinity than with other cells or substances. An antibody "specifically binds" or "preferredly binds" to a target if it binds with higher affinity, avidity, more easily, and / or for a longer period than it binds to other substances. Reading this definition, it is also understood that, for example, an antibody (or part or epitope) that specifically or preferentially binds to a first target may or may not specifically or preferentially bind to a second target. Thus, "specific binding" or "preferred binding" is not necessarily exclusive, although it may include such binding. An anti-CDH11 antibody or its antigen-binding fragment that specifically binds to human CDH11 is provided herein. Anti-CDH11 antibodies or their antigen-binding fragments that specifically bind to mouse CDH11 are provided herein. In several embodiments, the anti-CDH11 antibodies or their CDH11-binding fragments do not cross-react with mouse CDH11. Anti-CDH11 antibodies or their antigen-binding fragments are provided herein, which do not significantly bind to one or more of CDH5, CDH6, CDH7, CDH8, CDH9, CDH18, CDH20, or CDH24.

[0127] In multiple embodiments, the fragment that binds to the anti-CDH11 antibody or CDH11 binds to a linear epitope. In multiple embodiments, the fragment that binds to the anti-CDH11 antibody or CDH11 binds to a non-linear epitope. In one embodiment, the fragment that binds to the anti-CDH11 antibody or CDH11 binds to its antigen with one, two, three, four, five, or six CDRs.

[0128] As used herein with respect to antibodies, the term “competing” means that the first antibody or its antigen-binding moiety binds to the epitope in a manner sufficiently similar to the binding of the second antibody or its antigen-binding moiety, such that the result of the first antibody’s binding to its homologous epitope is detectably reduced in the presence of the second antibody compared to the binding of the first antibody in the absence of the second antibody. Alternatively, the binding of the second antibody to its epitope may be detected as well as in the presence of the first antibody, but this is not required. In other words, the first antibody can inhibit the binding of the second antibody to its respective epitope without the second antibody inhibiting the first antibody’s binding to that epitope. However, if each antibody detectably inhibits the binding of the other antibody to its homologous epitope or ligand, whether to the same degree, more, or less, the antibodies are said to “cross-compete” with each other for the binding of their respective epitopes. Both competing antibodies and cross-competing antibodies are disclosed herein. Regardless of the mechanism by which such competition or cross-competition occurs (e.g., steric hindrance, conformational change, or binding to a common epitope or part thereof), those skilled in the art will understand, based on the teachings herein, that such competing and / or cross-competitive antibodies may be included and useful in the methods disclosed herein.

[0129] Provided herein are anti-CDH11 antibodies or fragments thereof that bind to CDH11, which compete with any of the anti-CDH11 antibodies or fragments thereof that bind to CDH11 disclosed herein.

[0130] A bispecific antibody comprising one of the CDR sequences, variable heavy chain sequences, or variable light chain sequences disclosed herein is provided herein. As used herein, a bispecific antibody refers to an antibody capable of simultaneously binding to two antigens.

[0131] antibody fragment As used herein, the terms “antigen-binding moiety” or “antigen-binding fragment” may be fragments comprising Fab, Fab', F(ab')2, Fd, Fv, domain antibodies (dAb, e.g., shark and camel antibodies), ScFv, maxi-bodies, mini-bodies, nano-bodies, intra-bodies, dia-bodies, tria-bodies, tetra-bodies, v-NAR and bis-scFv, or polypeptides containing at least a specific portion of an immunoglobulin sufficient to confer specific antigen binding to the polypeptide.

[0132] For example, antibody fragments can be prepared by cleaving intact antibodies or by recombinant means. See, for general information, Fundamental Immunology, Ch. 7 (which is incorporated herein by reference in its entirety, Paul, W., ed., 2nd ed. Raven Press, NY (1989)). Antigen-binding fragments can be produced by recombinant DNA technology, by enzymatic or chemical cleavage of intact antibodies, or by molecular biological techniques.

[0133] In some embodiments of the aspects described herein, the antibodies disclosed herein are Fab fragments comprising, or essentially comprising, a variable (VL) and a constant (CL) domain of the light chain and a variable (VH) domain and a first constant (CH1) domain of the heavy chain.

[0134] In some embodiments of the aspects described herein, the antibody fragment is a Fab' fragment, which refers to a Fab fragment having one or more cysteine ​​residues at the C-terminus of the CH1 domain.

[0135] In some embodiments of the aspects described herein, the antibody fragment is an Fd fragment containing or essentially consisting of VH and CH1 domains.

[0136] In some aspects of the aspects described herein, the antibody fragment is an Fd' fragment comprising VH, a CH1 domain, and one or more cysteine ​​residues at the C-terminus of the CH1 domain.

[0137] A single-chain Fv or scFv antibody fragment contains, or essentially consists of, the VH and VL domains of a given antibody, such that the following domains are present in a single polypeptide chain. Generally, the Fv polypeptide further contains a polypeptide linker between the VH-VL domains, which allows the scFV to form a structure desirable for antigen binding. Therefore, in some aspects of the aspects described herein, the antibody fragment is an Fv fragment containing, or essentially consisting of, the VL and VH domains of a single arm of an antibody.

[0138] In some embodiments of the aspects described herein, the antibody fragment is a diabody comprising two antigen-binding sites, each containing a heavy chain variable domain (VH) connected to a light chain variable domain (VL) in the same polypeptide chain.

[0139] In some embodiments of the aspects described herein, the antibody fragment is a dAb fragment containing or essentially having a VH domain.

[0140] In some embodiments of the aspects described herein, the antibody fragment is an F(ab')2 fragment comprising a bivalent fragment containing two Fab' fragments linked by a disulfide bridge at a hinge region.

[0141] In some aspects of the configuration described herein, the antibody fragment is a linear antibody comprising a pair of tandem Fd segments (VH-CH1-VH-CH1) that form a pair of antigen-binding regions together with a complementary light chain polypeptide.

[0142] Those skilled in the art can utilize various techniques developed and available for the production of antibody fragments. Conventionally, these fragments have been induced via proteolytic digestion of intact antibodies. However, F(ab')2 fragments can be isolated directly from recombinant host cell cultures. Other techniques for the production of antibody fragments will be apparent to practitioners of the art. In another embodiment, the choice of antibody fragment is a single-chain Fv fragment (scFv). See, for example, WO 93 / 16185. Alternatively, these fragments can also be produced directly by recombinant host cells. For example, antibody fragments can be isolated from antibody phage libraries discussed herein. In another approach, Fab'-SH fragments can be recovered directly from Escherichia coli and chemically linked to form F(ab')2 fragments (Carter et al., 1992).

[0143] Antibody conjugate In some embodiments, the antibodies or antigen-binding fragments disclosed herein may include, but are not limited to, modifications such as glycosylation, acetylation, pegylation, phosphorylation, amidation, derivatization with known protecting / blocking groups, proteolytic cleavage, and linkage to cellular ligands or other proteins. Processes for producing chemical modifications are known in the art and may include, but are not limited to, specific chemical cleavage, acetylation, formylation, and metabolic synthesis of tunicamycin. In addition, the molecules may contain one or more non-classical amino acids.

[0144] In some aspects of the aspects described herein, an antibody or its antigen-binding fragment is conjugated to a functional portion. Examples of useful functional portions include, but are not limited to, blocking portions, detectable portions, diagnostic portions, targeting portions, and therapeutic portions.

[0145] Exemplary blocking moieties include portions with sufficient steric bulk and / or charge such that reduced glycosylation occurs, for example, by blocking the ability of glycosidase to glycosylate an antibody or its antigen-binding fragment. Blocking moieties may additionally or alternatively reduce effector function, for example, by inhibiting the ability of an Fc region to bind to a receptor or complementary protein. Preferred blocking moieties include cysteine ​​adducts and PEG moieties.

[0146] In one embodiment, the blocking portion is cysteine, preferably cysteine ​​associated with free cysteine, for example, during or after translation of an Fc-containing polypeptide in cell culture. Other blocking cysteine ​​adducts include cystine, mixed disulfide adducts, or disulfide linkages.

[0147] In another embodiment, the blocking moiety is a polyalkylene glycol moiety, for example, a PEG moiety, preferably a PEG-maleimide moiety. Preferred PEGylated moieties (or related polymers) may be, for example, polyethylene glycol ("PEG"), polypropylene glycol ("PPG"), polyoxyethylated glycerol ("POG"), and other polyoxyethylated polyols, polyvinyl alcohol ("PVA"), and other polyalkylene oxides, polyoxyethylated sorbitol, or polyoxyethylated glucose. The polymer may be a homopolymer, a random or blocked copolymer, a terpolymer based on the above monomers, linear or branched, substituted or unsubstituted, as long as it has at least one active sulfone moiety. The polymer moieties may be of any length or molecular weight, but these characteristics can affect the biological properties. In pharmaceutical applications, the average molecular weight of polymers particularly useful for reducing clearance rates is in the range of 2,000 to 35,000 daltons. In addition, when two groups are linked one at each end of a polymer, the length of the polymer can affect the effective distance between the two groups and other spatial relationships. Therefore, those skilled in the art can optimize or impart desired biological activity by varying the length of the polymer. For several reasons, PEG is useful for biological applications. PEG is typically colorless, odorless, water-soluble, heat-stable, inert to many chemical agents, does not hydrolyze, and is non-toxic. Pegylation can improve the pharmacokinetic performance of a molecule by increasing its apparent molecular weight. An increase in apparent molecular weight reduces the rate of clearance from the body after subcutaneous or systemic administration. In many cases, pegylation can reduce antigenicity and immunogenicity. In addition, pegylation can increase the solubility of biologically active molecules.

[0148] Examples of detectable portions for the detection of antibodies and their antigen-binding fragments disclosed herein include fluorescent moieties or labels, agents for image analysis, radioisotope moieties, radiopaque moieties, etc., including detectable labels such as biotin, fluorophores, chromophores, spin resonance probes, or radiolabels. Exemplary fluorophores include fluorescent dyes (e.g., fluorescein, rhodamine, etc.) and other luminescent molecules (e.g., luminol). Fluorophores may be environmentally sensitive, such that their fluorescence changes when located near one or more residues in a modified protein that undergo a structural change upon binding to a substrate (e.g., a dansyl probe). Exemplary radiolabels include one or more low-sensitivity nuclear ( 13 C, 15 N, 2 H, 125 I, 123 I, 99 Tc, 43 K, 52 Fe, 67 Ga, 68 Ga, 111 It contains low molecules containing atoms (such as In). Other useful parts are known in the art.

[0149] Examples of therapeutic components include, but are not limited to, anti-inflammatory agents, nonsteroidal anti-inflammatory drugs, steroids, disease-modifying antirheumatic agents (DMARDs), or recombinant proteins, and any such anti-inflammatory agent, nonsteroidal anti-inflammatory drug, steroid, disease-modifying antirheumatic agent (DMARD), or recombinant protein disclosed herein.

[0150] The functional parts may also have one or more of the above functions.

[0151] To increase the half-life of an antibody or polypeptide containing the amino acid sequences described herein, salvage receptor-binding epitopes can be attached to the antibody or its antigen-binding fragment (particularly the antibody fragment), as described, for example, in U.S. Patent No. 5,739,277. The term “salvage receptor-binding epitope” may refer to an epitope in the Fc region of an IgG molecule (e.g., IgG1, IgG2, IgG3, or IgG4) that plays a role in increasing the in vivo serum half-life of the IgG molecule (e.g., Ghetie et al., 18 Ann. Rev. Immunol. 739 (2000)). Antibodies having substitutions in their Fc region and exhibiting increased serum half-life are also described in WO 00 / 42072, WO 02 / 060919; Shields et al., 276 J. Biol. Chem. 6591 (2001); Hinton, 279 J. Biol. This is described in Chem. 6213-6216 (2004). For example, a nucleic acid molecule encoding a salvage receptor-binding epitope can be in-frame ligated to a nucleic acid encoding a polypeptide sequence described herein, thereby the fusion protein expressed by the manipulated nucleic acid molecule contains the salvage receptor-binding epitope and the polypeptide sequence described herein. In another embodiment, the serum half-life can also be increased, for example, by attaching other polypeptide sequences.

[0152] Other types of functional components are known in the art and can be readily used in the methods and compositions of this disclosure based on the teachings contained herein.

[0153] nucleic acids and vectors Nucleic acids encoding antibodies or antigen-binding fragments disclosed herein, as well as vectors, host cells, and expression systems, are also provided herein. As used herein, the term “nucleic acid” refers to a polymer of nucleotides of any length, either ribonucleotides or deoxyribonucleotides. Therefore, the term includes, but is not limited to, single-stranded, double-stranded, or multi-stranded DNA or RNA, genomic DNA, cDNA, DNA-RNA hybrids, or polymers containing purine and pyrimidine bases, or other natural, chemically or biochemically modified, unnatural, or derivatized nucleotide bases.

[0154] Nucleic acids encoding CDRs or variable-chain sequences disclosed herein are provided herein. Nucleic acids encoding variable-chain sequences containing sequences identical to at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% of any one of SEQ ID NO: 62 to 77 are provided herein. Nucleic acids encoding variable-chain sequences containing sequences identical to at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% of any one of SEQ ID NO: 78 to 93 are provided herein.

[0155] Nuclei containing sequences identical to any one of SEQ ID NO: 94-127 by at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% are provided herein. Nucleic acids or sets of nucleic acids containing two or more sequences identical to any one of SEQ ID NO: 94-127 by at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% are also provided herein.

[0156] In some embodiments, nucleic acid sequences encoding heavy chain variable regions are provided, each nucleic acid sequence comprising a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to any one of SEQ ID NO: 94-110. In some embodiments, nucleic acid sequences encoding light chain variable regions are provided, each nucleic acid sequence comprising a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to any one of SEQ ID NO: 111-127. In some embodiments, nucleic acid sequences encoding heavy chain variable regions are provided, each nucleic acid sequence comprising a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to any one of SEQ ID NO: 111-127.

[0157] Nucleic acids encoding one or more (or variants thereof) of the amino acid sequences in Table 1 or Table 2 are provided herein. A set of nucleic acids encoding one (or variant thereof) of the variable heavy-chain and light-chain pairs disclosed in Table 2 is provided herein.

[0158] The nucleic acids encoding antibodies or antigen-binding fragments disclosed herein may be recombinantly produced chimeric nucleic acid molecules comprising, for example, DNA, cDNA, RNA, synthetically produced DNA or RNA, or any of these polynucleotides, either alone or in combination. For example, an expression vector or set of expression vectors is provided, comprising a polynucleotide sequence encoding an antibody or antigen-binding fragment described herein, operably linked to an expression regulatory sequence suitable for expression in eukaryotic and / or prokaryotic host cells.

[0159] The term "vector" refers to a nucleic acid molecule capable of transporting another nucleic acid to which it is ligated. A "vector" may include, but is not limited to, viral vectors, plasmids, RNA vectors, or linear or circular DNA or RNA molecules that may consist of chromosomal, non-chromosomal, semi-synthetic, or synthetic nucleic acids. In some embodiments, the vectors employed are capable of autonomous replication (episome vectors) and / or capable of expressing the nucleic acid to which they are ligated (expression vectors). Numerous suitable vectors are known to those skilled in the art and are commercially available. Viral vectors include negative-strand RNA viruses such as retroviruses, adenoviruses, parvoviruses (e.g., adeno-associated viruses, AAV), coronaviruses; orthomyxoviruses (e.g., influenza virus), rhabdoviruses (e.g., rabies and vesicular stomatitis viruses), and paramyxoviruses (e.g., measles and Sendai viruses); 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, fowlpox, and canarypox). Other viruses include, for example, Norwalk virus, togavirus, flavivirus, reovirus, papovavirus, hepadnavirus, and hepatitis viruses. Examples of retroviruses include avian leukemia sarcoma virus, mammalian type C, B, and D viruses, the HTLV-BLV group, lentiviruses, and spumaviruses.

[0160] Vectors comprising nucleic acids encoding antibodies or antigen-binding fragments disclosed herein are also provided herein.

[0161] A vector comprising nucleic acids encoding variable-chain sequences disclosed herein is provided herein. A nucleic acid encoding a variable-chain sequence comprising a sequence identical to at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% of any one of SEQ ID NO: 62 to 77 is provided herein. A vector comprising nucleic acids encoding a variable-chain sequence comprising a sequence identical to at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% of any one of SEQ ID NO: 78 to 93 is provided herein.

[0162] A vector comprising a nucleus containing a sequence identical to any one of SEQ ID NO: 94-127 by at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% is provided herein. A vector comprising a nucleic acid or set of nucleic acids containing two or more sequences identical to any one of SEQ ID NO: 94-127 by at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% is also provided herein.

[0163] In some embodiments, a vector is provided containing a nucleic acid sequence encoding a heavy chain variable region, the nucleic acid sequence containing a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to any one of SEQ ID NO: 94-110. In some embodiments, a vector is provided containing a nucleic acid sequence encoding a light chain variable region, the nucleic acid sequence containing a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to any one of SEQ ID NO: 111-127. In some embodiments, a vector is provided containing a nucleic acid sequence encoding a heavy chain variable region, the nucleic acid sequence containing one of SEQ ID NO: 94-110. In some embodiments, a vector is provided containing a nucleic acid sequence encoding a light chain variable region, the nucleic acid sequence containing one of SEQ ID NO: 111-127.

[0164] Vectors comprising nucleic acids encoding one or more (or variants thereof) of any amino acid sequences in Table 1 or Table 2 are provided herein. Sets of vectors comprising nucleic acids encoding any (or variants thereof) of any of the variable heavy-chain and light-chain pairs disclosed in Table 2 are provided herein.

[0165] In some embodiments, a vector or set of vectors is provided, comprising (1) a nucleic acid sequence containing one of SEQ ID NO: 94 to 110, and (2) a nucleic acid sequence containing one of SEQ ID NO: 111 to 127.

[0166] In one aspect, the following: (a) SEQ ID NO: 94 and / or SEQ ID NO: 111; (b) SEQ ID NO: 95 and / or SEQ ID NO: 112; (c) SEQ ID NO:96 and / or SEQ ID NO:113; (d) SEQ ID NO:97 and / or SEQ ID NO:114; (e) SEQ ID NO:98 and / or SEQ ID NO:115; (f) SEQ ID NO: 99 and / or SEQ ID NO: 116; (g) SEQ ID NO: 100 and / or SEQ ID NO: 117; (h) SEQ ID NO: 101 and / or SEQ ID NO: 118; (i) SEQ ID NO: 102 and / or SEQ ID NO: 119; (j) SEQ ID NO: 103 and / or SEQ ID NO: 120; (k) SEQ ID NO: 104 and / or SEQ ID NO: 121; (l) SEQ ID NO: 105 and / or SEQ ID NO: 122; (m) SEQ ID NO: 106 and / or SEQ ID NO: 123; (n) SEQ ID NO: 107 and / or SEQ ID NO: 124; (o) SEQ ID NO: 108 and / or SEQ ID NO: 125; (p) SEQ ID NO: 109 and / or SEQ ID NO: 126; or (q) SEQ ID NO: 110 and / or SEQ ID NO: 127 A vector or set of vectors is provided, including the following.

[0167] Various expression vectors have been developed to efficiently synthesize antibodies or their antigen-binding fragments in prokaryotic cells such as bacteria, and in eukaryotic cell systems, including but not limited to developed culture systems of yeast and mammalian cells. The vectors may include chromosomal, non-chromosomal, and synthetic DNA sequence segments. Cells containing expression vectors for expressing the disclosed antibodies or their antigen-binding fragments are also provided.

[0168] In one embodiment, a host cell containing a nucleic acid molecule or a vector described herein is provided. The host cell can be isolated. In several embodiments, the cell is a mammalian, insect cell, fungal cell, or bacterial cell.

[0169] Antibody production In one embodiment, a method for producing an antibody or antigen-binding fragment disclosed herein is provided, comprising the step of culturing cells containing one or more nucleic acid molecules encoding the antibody or antigen-binding fragment, under conditions in which the antibody or antigen-binding fragment is produced by a host cell.

[0170] The following steps: (a) A step of culturing cells containing one or more nucleic acid molecules encoding the anti-CDH11 antibody or the CDH11-binding fragment thereof, under conditions in which the host cell produces the CDH11 antibody or the CDH11-binding fragment thereof as disclosed herein; (b) Optionally, a step of substantially isolating an anti-CDH11 antibody or a fragment thereof that binds to CDH11. Methods for producing anti-CDH11 antibodies or fragments thereof that bind to CDH11, as disclosed herein, are provided herein.

[0171] The antibodies or antigen-binding fragments disclosed herein are typically produced by recombinant expression. Nucleic acids encoding the light chain variable region and the heavy chain variable region may optionally be ligated to the constant region and inserted into the same expression vector. Alternatively, the nucleic acids encoding the light chain variable region and the heavy chain variable region may optionally be ligated to the constant region and inserted into different expression vectors. The expression vector may further include one or more expression regulatory sequences, including, but not limited to, a promoter (e.g., a homogeneous or heterogeneous promoter), a signal sequence, an enhancer element, and a transcription termination sequence. Preferably, the expression regulatory sequence is a eukaryotic promoter system in the vector that can transform or transfect eukaryotic host cells. Typically, the host is maintained under conditions suitable for high-level expression of the nucleotide sequence, and after the vector is incorporated into a suitable host, the cross-reactive antibody is recovered and purified.

[0172] Generally, expression vectors include a selection marker (e.g., ampicillin resistance, hygromycin resistance, tetracycline resistance, or neomycin resistance) that allows for the detection of cells transformed with the desired DNA sequence.

[0173] The host used to express the antibodies or antigen-binding fragments disclosed herein may be a prokaryotic host or a eukaryotic host. Examples of suitable hosts include bacterial hosts, or eukaryotic hosts including yeast, insect, fungal, avian, and mammalian cells, either in vivo or in situ, or host cells of mammalian, insect, avian, or yeast origin. Mammalian cells or tissues may be of human, primate, hamster, rabbit, rodent, cattle, pig, sheep, horse, goat, dog, or cat origin, but any other mammalian cells may also be used.

[0174] Examples of bacterial hosts that can be used to express the antibodies and antigen-binding fragments disclosed herein include Escherichia coli, bacilli such as Bacillus subtilus, and other Enterobacteriaceae such as Salmonella, Serratia, and various Pseudomonas species.

[0175] Yeast can also be used as a host for expressing the antibodies, antigen-binding fragments, or fusion proteins disclosed herein. Saccharomyces and Pichia are exemplary yeast hosts, and suitable vectors having, as desired, expression regulatory sequences (e.g., promoters), origins of replication, termination sequences, etc., are used. Typical promoters include 3-phosphoglycerate kinase and other glycolytic enzymes. Inducible yeast promoters include, among others, alcohol dehydrogenase, isocytochrome C, and promoters derived from enzymes that play a role in the utilization of methanol, maltose, and galactose.

[0176] Mammalian cells in culture may also be used as host cells for expressing the antibodies, antigen-binding fragments, or fusion proteins disclosed herein. Examples of suitable host cell lines known in the art that can secrete heterologous proteins (e.g., intact immunoglobulins) include CHO cell lines, various COS cell lines, HeLa cells, 293 cells, myeloma cell lines, transformed B cells, and hybridomas. Expression vectors for these cells may include origins of replication, promoters, enhancers, and regulatory sequences such as necessary processing information sites, e.g., ribosome binding sites, RNA splice sites, and / or transcriptional terminator sequences. Examples of regulatory sequences include SV40, adenovirus, bovine papillomavirus, and cytomegalovirus.

[0177] The antibodies or antigen-binding fragments disclosed herein can be expressed using a single expression construct or vector, or multiple expression constructs or vectors (e.g., two or three expression constructs). If the heavy and light chains of the antibody are cloned into separate expression vectors, these vectors are co-transfected to obtain the expression and assembly of intact immunoglobulin. Once expressed, the whole antibody, its dimers, individual light and heavy chains, or other immunoglobulin forms disclosed herein can be purified according to standard procedures in the art, including ammonium sulfate precipitation, affinity column chromatography, column chromatography, HPLC purification, and gel electrophoresis (see, for example, Scopes, Protein Purification (Springer-Verlag, NY, (1982))). For pharmaceutical use, substantially pure immunoglobulin with at least about 90–95% homogeneity is preferred, with 98–99% or greater homogeneity being most preferred.

[0178] The antibodies or antigen-binding fragments disclosed herein can be prepared by any method known in the art.

[0179] Pharmaceutical composition In one embodiment, a pharmaceutical composition is provided comprising an anti-CDH11 antibody or a fragment thereof that binds to CDH11, as disclosed herein.

[0180] In one embodiment, a pharmaceutical composition is provided comprising an antibody or antigen-binding fragment disclosed herein and a pharmaceutically acceptable excipient. The pharmaceutically acceptable excipient may be a pharmaceutically acceptable material, composition or vehicle involved in carrying or transporting the therapeutic compound for administration to a subject, and may be, for example, a liquid or solid filler, diluent, manufacturing aid (e.g., lubricant, magnesium talc, calcium stearate or zinc stearate, or steric acid), solvent, or encapsulating material, or a bulking agent, salt, surfactant, and / or preservative. Some examples of materials that can act as pharmaceutically acceptable excipients include: sugars, e.g., lactose, glucose, and sucrose; starches, e.g., corn starch and potato starch; cellulose and its derivatives, e.g., sodium carboxymethylcellulose, ethylcellulose, and cellulose acetate; gelatin; talc; waxes; oils, e.g., peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil; glycols, e.g., ethylene glycol and propylene glycol; polyols, e.g., glycerin, sorbitol, mannitol, and polyethylene glycol; esters, e.g., ethyl oleate and ethyl laurate; agar; buffers; water; isotonic salines, pH buffered solutions, and other non-toxic, suitable substances used in pharmaceutical formulations.

[0181] The compositions or pharmaceutical compositions disclosed herein, comprising antibodies or antigen-binding fragments, include stabilizers to prevent loss of protein activity or structural integrity due to denaturation, oxidation, or aggregation over a period of time during storage and transport before use. The compositions or pharmaceutical compositions may include one or more of any combinations of salts, surfactants, and pH and osmotic pressure modifiers, such as sugars, which contribute to overcoming aggregation problems. When the compositions or pharmaceutical compositions disclosed herein are used as injectable preparations, the compositions may have a pH value in a nearly neutral pH range. In some embodiments, the level of surfactants is minimized to avoid bubbles in the formulation, which are detrimental to injection into the subject. In some embodiments, the compositions or pharmaceutical compositions are in liquid form, stably retaining high concentrations of biologically active antibodies in solution and suitable for inhalation or parenteral administration. In some embodiments, the compositions or pharmaceutical compositions are suitable for intravenous, intramuscular, intraperitoneal, intradermal, and / or subcutaneous injection. In some embodiments, the compositions or pharmaceutical compositions are in liquid form, minimizing the risk of bubble formation and anaphylactic-like side effects. In one embodiment, the composition or pharmaceutical composition is isotonic. In one embodiment, the composition or pharmaceutical composition has a pH of 6.8 to 7.4.

[0182] In one embodiment, the antibody or antigen-binding fragment disclosed herein is lyophilized and / or freeze-dried and reconstituted for use.

[0183] The antibodies or antibody fragments, or compositions, or pharmaceutical compositions described herein can be lyophilized or provided in any suitable form, including but not limited to injectable or inhalable solutions, gels, and tablets.

[0184] Methods using the antibodies described herein In multiple embodiments, the anti-CDH11 antibody or its fragment bound to CDH11 disclosed herein interferes with CDH11:CDH11 binding between cells. In multiple embodiments, the anti-CDH11 antibody or its fragment bound to CDH11 disclosed herein interferes with CDH11:CDH11 binding between HSCs. In multiple embodiments, the anti-CDH11 antibody or its fragment bound to CDH11 disclosed herein interferes with CDH11:CDH11 binding between HSCs and other mesenchymal-derived CDH11 + It interferes with CDH11:CDH11 binding between fibrotic cells.

[0185] In several embodiments, the anti-CDH11 antibody or fragment thereof that binds to CDH11 as disclosed herein is CDH11 + It suppresses the phenotype of cellular fibrosis.

[0186] A method for inhibiting intercellular CDH11-CDH11 interactions is provided herein, comprising the step of bringing cells into contact with an anti-CDH11 antibody or a fragment thereof that binds to CDH11, or a pharmaceutical composition comprising an anti-CDH11 antibody or a fragment thereof that binds to CDH11, as disclosed herein.

[0187] A method for inhibiting CDH11-CDH11 interaction in a subject in need is provided herein, comprising the step of administering to a subject an anti-CDH11 antibody or a fragment thereof that binds to CDH11, or a pharmaceutical composition comprising an anti-CDH11 antibody or a fragment thereof that binds to CDH11, as disclosed herein.

[0188] Methods of using a pharmaceutical composition comprising an anti-CDH11 antibody or a fragment thereof bound to CDH11, or an anti-CDH11 antibody or a fragment thereof bound to CDH11, for the treatment of a disease in a subject where such treatment is necessary are provided herein. The use of a pharmaceutical composition comprising an anti-CDH11 antibody or a fragment thereof bound to CDH11, or an anti-CDH11 antibody or a fragment thereof bound to CDH11, for the treatment of a disease is provided herein. An anti-CDH11 antibody or a fragment thereof bound to CDH11, for use in the manufacture of a pharmacopoeia for the treatment of a disease is provided herein.

[0189] In several embodiments, the subject is an animal. In several embodiments, the subject is a mammal. In some embodiments, the subject is a non-human mammal. In several embodiments, the subject is a non-primate (e.g., cattle, buffalo, pigs, horses, cats, dogs, rats, mice, guinea pigs, sheep, goats, etc.) or a primate (e.g., monkeys and humans). In one embodiment, the subject is a human. Individuals and patients are also subjects in this specification.

[0190] As used herein, the terms “treat,” “treated,” “treating,” or “treatment” refer to therapeutic treatments whose purpose is to delay (alleviate) an undesirable physiological condition, disorder, or disease, or to obtain a beneficial or desirable clinical outcome. For the purposes of the present invention, beneficial or desirable clinical outcomes include, but are not limited to, symptom relief; reduction of the severity of a condition, disorder, or disease; stabilization of the condition, disorder, or disease (i.e., no worsening); delay of the onset or slowing of the progression of a condition, disorder, or disease; recovery of the condition, disorder, or disease; and remission (partial or complete), whether detectable or undetectable; or enhancement or improvement of the condition, disorder, or disease. Treatment includes eliciting a clinically significant response without excessive side effects. Treatment also includes extending survival compared to the survival expected without treatment. Terms such as "prevent" and "prevention" refer to acting before the onset of an obvious disease or disorder to prevent its development, minimize its severity, or delay its progression.

[0191] A method for treating fibrosis in a subject in need is provided herein, comprising the step of administering to a subject an anti-CDH11 antibody or a fragment thereof bound to CDH11 disclosed herein, or a pharmaceutical composition comprising an anti-CDH11 antibody or a fragment thereof bound to CDH11 disclosed herein. An anti-CDH11 antibody or a fragment thereof bound to CDH11 disclosed herein for use in the treatment of fibrosis in a subject in need is provided herein. Use of an anti-CDH11 antibody or a fragment thereof bound to CDH11 disclosed herein in the manufacture of a pharmaceutical or in the manufacture of a pharmaceutical composition for treating fibrosis in a subject in need is provided herein. In several embodiments, an anti-CDH11 antibody or a fragment thereof bound to CDH11 reverses or inhibits the progression of disease in fibrosis. In multiple embodiments, fibrosis is hepatic fibrosis, pulmonary fibrosis, renal fibrosis, cardiac fibrosis, myelofibrosis, pancreatic fibrosis, cutaneous fibrosis, intestinal fibrosis, post-myocardial infarction fibrosis, post-replacement fibrosis, perivascular fibrosis, or articular fibrosis. In one embodiment, fibrosis is hepatic fibrosis. In multiple embodiments, the subject has a hepatitis virus infection, non-alcoholic fatty liver disease (NASH), a hereditary metabolic disorder, a cholestatic disorder, or an immune disorder. In multiple embodiments, hepatic fibrosis is associated with increased alcohol intake or excessive vitamin A intake. In multiple embodiments, hepatic fibrosis is associated with autoimmune diseases of the liver or bile ducts (including but not limited to autoimmune hepatitis, primary biliary cholangitis, or primary sclerosing cholangitis), drug toxicity, liver graft rejection, or genetic disorders of the liver (including but not limited to Wilson's disease or hemochromatosis), or bile ducts (including but not limited to biliary atresia). In one embodiment, fibrosis is intestinal fibrosis. In multiple embodiments, the subject has inflammatory bowel disease (IBD), Crohn's disease, or colitis. In one embodiment, pulmonary fibrosis is induced as idiopathic pulmonary fibrosis or toxic injury (including, but not limited to, bleomycin and silicosis). In multiple embodiments, the subject has scleroderma or systemic scleroderma. In multiple embodiments, the subject has keloid scarring.

[0192] A method for treating rheumatoid arthritis in a subject where such treatment is needed is provided herein, comprising the step of administering to a subject an anti-CDH11 antibody or a fragment thereof bound to CDH11 as disclosed herein, or a pharmaceutical composition comprising an anti-CDH11 antibody or a fragment thereof bound to CDH11 as disclosed herein. An anti-CDH11 antibody or a fragment thereof bound to CDH11 as disclosed herein for use in the treatment of rheumatoid arthritis in a subject where such treatment is needed is provided herein. Use of an anti-CDH11 antibody or a fragment thereof bound to CDH11 as disclosed herein in the manufacture of a pharmaceutical, or use in the manufacture of a pharmaceutical composition for treating rheumatoid arthritis in a subject where such use is needed is provided herein.

[0193] The antibodies or antigen-binding fragments disclosed herein may be administered together with one or more additional therapeutic agents to induce disease remission / control, or to maximize the therapeutic response without further impairing the systemic immune response to the microorganism. Suitable additional therapeutic agents in this regard include, but are not limited to, anti-TNFα agents.

[0194] In several embodiments, the anti-CDH11 antibody or a fragment thereof bound to CDH11 disclosed herein, or a pharmaceutical composition comprising the disclosed anti-CDH11 antibody or a fragment thereof bound to CDH11, is administered alone or in combination with one or more other therapeutic agents (e.g., anti-inflammatory agents or metabolites) or agents that inhibit mesenchymal cell activation through growth factor receptor inhibition or depletion of fibrinoplastic cells.Suitable anti-inflammatory agents useful for the treatment of inflammatory joint diseases, particularly rheumatoid arthritis (RA), which can be administered in combination with the anti-CDH11 antibodies or fragments thereof that bind to CDH11 disclosed herein include: (i) nonsteroidal anti-inflammatory drugs (NSAIDs); for example, detoprofen, diclofenac, diflunisal, etodolac, fenoprofen, flurbiprofen, ibuprofen, indomethacin, ketoprofen, meclofenamete, mefenamic acid, meloxicam, nabumethionine, naproxen sodium, oxaprozin, piroxicam, sulindac, tolme (ii) Steroids (e.g., cortisone, dexamethasone, hydrocortisone, methylprednisolone, prednisolone, prednisone, triamcinolone); (iii) DMARDs, i.e., disease-modifying antirheumatic drugs (e.g., cyclosponne, azathioprine, methotrexate, leflunomide, cyclophosphamide, hydroxychloroquine, sulfamethasone); (ii) Steroids (e.g., cortisone, dexamethasone, hydrocortisone, methylprednisolone, prednisolone, prednisone, triamcinolone); (iii) DMARDs, i.e., disease-modifying antirheumatic drugs (e.g., cyclosponne, azathioprine, methotrexate, leflunomide, cyclophosphamide, hydroxychloroquine, sulfamethasone Rufasalazine, D-penicillamine, minocycline, and gold); or (iv) recombinant proteins (e.g., ENBREL® (etanercept, soluble TNF receptor), REMICADE® (infliximab, chimeric monoclonal anti-TNF antibody), ORENCIAO (abatabacept, soluble CTLA4 receptor), ACTEMR® (tocilizumab, monoclonal antibody against IL-6 receptor), and RITUXAN® (rituximab, monoclonal antibody against CD20) (v) antibodies; or metabolites used to treat non-alcoholic steatohepatitis (NASH) (including thyroid hormone β-agonists, FXR agonists, inhibitors of adipogenesis, inhibitors of transforming growth factor β activation (e.g., integrins or other small molecule antagonists), insulin resistance improvers (e.g., GLP-1, GIP or glucagon agonists, SGLT antagonists)); or (vi) inhibitors of mesenchymal cell activation, including but not limited to PDGF, EGF, or endothelin receptor inhibitors, or CAR-T cells that deplete fibrogenic cells.

[0195] Assay Device and Method of Detection Devices and methods for the detection of CDH11 are provided herein. In multiple embodiments, the devices provided herein include one or more of the anti-CDH11 antibodies or fragments that bind to CDh11 disclosed herein. In multiple embodiments, the devices provided herein include one or more of the antibodies disclosed herein.

[0196] In multiple embodiments, the antigen to be detected is directly immobilized on the surface of a multi-well microtiter and then contacted with an anti-CDH11 antibody or an antigen-binding fragment thereof disclosed herein that comprises a detection moiety.

[0197] In multiple embodiments, the antigen to be detected is directly immobilized on the surface of a multi-well microtiter and then contacted with an anti-CDH11 antibody or an antigen-binding fragment thereof disclosed herein. The anti-CDH11 antibody or an antigen-binding fragment thereof is detected by a secondary antibody that binds to the anti-CDH11 antibody or an antigen-binding fragment thereof disclosed herein that comprises a detection moiety.

[0198] In multiple embodiments, the anti-CDH11 antibodies or antigen-binding fragments disclosed herein are used in sandwich ELISA or a variant thereof. In sandwich ELISA, the analyte to be measured is bound between two primary antibodies that each detect a different epitope of the antigen. The first primary antibody, which is the capture antibody, is often immobilized on the surface. The second primary antibody often comprises a detection moiety. However, the second primary antibody may not be labeled, in which case it can be detected by a secondary antibody that binds to the second primary antibody and comprises a detection moiety. In multiple embodiments, the anti-CDH11 antibodies or antigen-binding fragments disclosed herein are used as the first primary antibody. In multiple embodiments, the anti-CDH11 antibodies or antigen-binding fragments disclosed herein are used as the second primary antibody.

[0199] Provided herein is a method for detecting CDH11, comprising the steps of contacting a biological sample with an anti-CDH11 antibody or a fragment thereof that binds to CDH11, and detecting the binding of the anti-CDH11 antibody or the fragment thereof that binds to CDH11 to CDH11. The binding of the anti-CDH11 antibody or the fragment thereof that binds to CDH11 to its antigen can be detected by various methods, including but not limited to: (1) attaching a reporting moiety to the anti-CDH11 antibody or the fragment thereof that binds to CDH11; or (2) contacting the CDH11 bound by the first anti-CDH11 antibody or the fragment thereof that binds to CDH11 with a second anti-CDH11 antibody or a fragment thereof that binds to CDH11 (which may further have a reporting moiety attached thereto); or (3) contacting the second anti-CDH11 antibody or the fragment thereof that binds to CDH11 with a third antibody that recognizes an antigen located on the second anti-CDH11 antibody or the fragment thereof that binds to CDH11, such as the constant region of the second anti-CDH11 antibody or the fragment thereof that binds to CDH11 (which may further have a reporting moiety attached thereto).

[0200] In certain embodiments, the reporting entity comprises an enzyme. In certain embodiments, the enzyme is horseradish peroxidase (HRP) or alkaline phosphatase (AP). In one embodiment, the reporting entity comprises gold nanoparticles.

[0201] The anti-CDH11 antibody or the fragment thereof that binds to CDH11 can be attached to a solid support of a device. In multiple embodiments, the solid support comprises nitrocellulose. In multiple embodiments, the device further comprises a fluid sample pad that sequentially precedes the first and second portions.

[0202] Also provided herein is a kit comprising one or more of the antibodies or antigen-binding fragments disclosed herein and one or more buffers.

[0203] It should be understood that the present invention is not limited to and may vary from the specific molecules, compositions, methodologies, or protocols described herein. Any methods and materials similar or equivalent to those described herein may be used in the implementation or testing of aspects of the present invention. It should be further understood that the disclosure of the present invention in this specification includes all possible combinations of such specific features. For example, if a specific feature is disclosed in the context of a particular aspect or form of the present invention or a particular claim, that feature may also be used, to the extent possible, in combination with other particular aspects and aspects of the present invention and / or in the context of other particular aspects and aspects of the present invention, and in the present invention in general.

[0204] In this specification, "and / or" as used, for example, with option A and / or option B, encompasses separate embodiments of (i) option A, (ii) option B, and (iii) option A + option B.

[0205] Where relating herein to a method comprising two or more defined steps, the defined steps may be performed in any order or simultaneously (unless the context excludes such possibility), and the method may include one or more other steps performed before any of the defined steps, between two defined steps, or after all of the defined steps (unless the context excludes such possibility).

[0206] All other patents and applications referenced herein are incorporated herein in their entirety by reference. Furthermore, if any definition or use of a term in a reference incorporated herein by reference conflicts with or contradicts a definition of that term provided herein, the definition provided herein shall prevail, and the definition in the reference shall not prevail.

[0207] To facilitate understanding of the present invention, specific embodiments are shown below. The following embodiments should not be read as limiting or defining the entire scope of the present invention. [Examples]

[0208] Example 1: Production of anti-CDH11 antibody CDH11 Structure Full-length human (ENST00000268603.8) and mouse (ENSMUST00000075190.4) CDH11 full-length and extracellular domain (ECD) CDS DNA were synthesized (GeneArt, ThermoFisher) and cloned into various vectors with and without C-terminal tags. For immunization, full-length human CDH11 was cloned untagged into the pcDNA3.1 mammalian expression vector (ThermoFisher). For flow cytometry-based screening, full-length human and mouse CDH11 were cloned into the pSelect-cGFP vector (InVivoGen) to produce full-length membrane-bound CDH11 protein with an intracellular GFP tag fused to the C-terminus. These constructs were transfected into Expi293 cells (ThermoFisher) and analyzed by flow cytometry. For proteins used in immunization and ELISA, human CDH11 ECD DNA, in which a human G1 Fc or mouse G2a Fc tag was recombinantly fused to the c-terminus of the CDH11 ECD protein domain, was cloned into a pFuse_Fc tagged vector. These constructs were used to transfect Expi293 cells for the production of recombinant Fc-tagged proteins secreted into the supernatant. The proteins were purified from the supernatant using Protein A / G.

[0209] To determine cross-reactivity with other type 2 cadherins, CDS DNA for eight different full-length human cadherins (CAD5, 6, 7, 8, 18, 20, and 24) was also synthesized and cloned into pSelect_cGFP. This produced c-terminally tagged recombinant GFP membrane-bound protein, which was used for flow cytometry as described above. Table 4 provides an overview of the DNA constructs.

[0210] (Table 4) DNA constructs. ECD = outer domain. X indicates the use of the vector for the stated purpose. TIFF2026515695000005.tif51152

[0211] Immunization and serum screening Two groups of VelocImmune® mice were immunized using various strategies (Figure 1).

[0212] One set (Ms# 139-143) was immunized twice with full-length human CDH11 DNA (cloned into a mammalian expression vector, pcDNA3.1), and then protein-immunized with a human CDH11 extracellular domain (ECD)_Fc fusion tag protein (Figure 1A). Two types of tagged proteins were used: one containing mouse Fc(mIgG2a) and the other containing human Fc(hIgG1). The mouse Fc-tagged protein was primarily used as a boosting immunogen, while the human Fc-tagged protein was used as a final boost to induce a CDH11-specific response.

[0213] In a similar manner, the second animal set (Ms#135-138) was given only protein immunization (Figure 1B).

[0214] Mouse serum from various post-immunization time points was screened by ELISA on human CDH11 ECD protein tagged with 5 μg / ml human IgG1-Fc (Figure 2A). Serum was also screened by flow on Expi293F cells (Invitrogen) transfected with GFP-tagged human and mouse full-length CDH11 constructs (Figure 2B).

[0215] Screening of fusions Over a six-month period, three fusions were performed on responsive mice, and all mice fused except for mice #138 and #139, which showed very low titers. Screening was performed using the hybridoma supernatant of Expi293F cells transfected with GFP-fused human CDH11. ELISA was also performed using human Fc(IgG1)-tagged human CDH11 ECD protein at 5 μg / ml (Figure 3). After two rounds of screening, 47 clones that showed a positive signal by flow or ELISA were identified (Figure 3, Table 6). For positive controls, fused serum (shown at a 1:1000 dilution) and commercially available anti-human CDH11, clone 16G5 (BioLegend) (at 1 μg / ml) were used. Normal mouse serum (at a 1:1000 dilution) was used as a negative control. Isotyping was performed using a standard mouse isotyping kit. Clones that showed flow reactivity were considered to have a higher priority than ELISA-positive clones due to the greater expectation of structural binding.

[0216] Example 2: Binding of hybridoma supernatant to human and rat primary fibroblast-like synovial cells (FLS) Human and rat primary FLS were used to determine the binding of hybridoma supernatants selected from fusions 1 and 2 by flow cytometry based on cell availability. Similar to the fusion serum (1:1000 dilution), commercially available cadherin control antibody 16G5 was used as a positive control (1 μg / ml). Normal mouse serum (1:1000 dilution) and irrelevant isotype control antibody (1 μg / ml). APC-conjugated goat anti-mouse IgG (Fc specific) (Jackson ImmunoResearch) was used as the secondary antibody.

[0217] Clones 2D2, 10E1, 10F1, 11D2, and 11D3 showed positive staining against both human and rat FLS (Figure 4). Clone 5A2 showed slight positivity against human synovial cells. Human FLS were not available during the screening of fusion 3 hybridomas, but for fusion 3 hybridomas, binding to rat primary FLS was determined. Two clones, namely 2A1 and 2B9, were reactive against rat FLS.

[0218] Example 3: Cross-reactivity of hybridoma supernatants against type 2 cadherin and mouse CDH11 To test the possibility of cross-reactivity against other human type 2 cadherins and mouse CDH11, hybridomas were screened by flow cytometry on Expi293F cells expressing one of eight different human type 2 cadherins (cadherin 5, 6, 8, 9, 18, 20, and 24) fused to GFP (Table 5). Hybridoma supernatants from CDH11-binding clones were screened by flow cytometry for binding to Expi293F HEK cells expressing different cadherins labeled with GFP at the C-terminus. The results are shown as relative percentage graphs for each clone. Commercially available anti-huCDH11 antibody 16G5, fusion serum, and normal mouse serum were used as controls.

[0219] Cross-reactivity was also determined using purified antibodies. Lead antibodies were purified from VelcoImmune-derived hybridoma supernatant and tested on Expi293 cells transiently transfected with either GFP-tagged human or mouse CAD11. Antibodies were stained at a concentration of 2.5 μg / ml and compared to mouse anti-human CAD11 control (clone 16G5), isotope control (mouse G1), or cells alone (mock transfection) by flow cytometry. For detection, anti-mouse IgG secondary antibody conjugated with APC fluorophores was added at a 1:1000 dilution. Mean fluorescence intensity (MFI) is reported (Figure 5).

[0220] The homology / identity of various cadherins with human CDH11 ranges from 40% for cadherin 5 to 66% for cadherin 8 (Table 6). Mouse CDH11 is highly homologous to human CDH11 in terms of amino acid sequence, sharing 97% amino acid identity, with most of the differences occurring in the membrane-proximal extracellular (EC) domains 3, 4, and 5.

[0221] The hybridomas primarily showed binding to either human and / or mouse CDH11, but not to other cadherins. Clones that showed cross-reactivity to other constructs were mainly ELISA-positive clones (e.g., 3F8, 3G8, 5C4, and 1G11 in fusion 1; 2F6, 3G6, and 17C1 in fusion 2; 2B9 and 5E4 in fusion 3), showed very low binding to all cadherins in flow (most of these were ELISA-positive clones), or were broadly reactive to all human cells, as shown in the overall list (3F8, 3G8, 5C4, 5E4). Clone 2D2 from fusion 1 showed slight positivity to cadherin 8, but was overwhelmingly positive to human and mouse CDH11. Only one clone, 5F12 from fusion 1, actually showed some cross-reactivity to another cadherin, cadherin 7.

[0222] Table 7 provides an overview of the clones tested.

[0223] (Table 5) Cross-reactivity of hybridoma supernatant with different type 2 cadherins HuCAD = human cadherin. MoCAD = mouse cadherin. Hybridoma supernatants from CDH11-binding clones were screened by flow cytometry for binding to Expi293F HEK cells expressing different cadherins labeled with GFP at the C-terminus. Commercial anti-huCDH11 antibody 16G5, fusion serum, and normal mouse serum were used as controls. TIFF2026515695000006.tif201146TIFF2026515695000007.tif103146

[0224] (Table 6) Homology of type 2 cadherins TIFF2026515695000008.tif42146

[0225] (Table 7) Summary of clones. Summary of binding analysis of the 47 clones generated. For ELISA, binding MFI or absorbance OD measurements were normalized and ranked against a commercially available control antibody (16G5). Binding was considered "low" if less than 25% of the control events occurred, positive if 25% or more of the control events occurred, and double-positive if within or above the control range (±10%). Clones within the range (±5%) of the negative control (normal mouse serum and mock cells) were considered negative. Interpretation of cross-reactivity to other type 2 cadherins, as well as the results of binding to primary FLS cells, are also included in the summary. Human FLS cells were available only for selected groups of clones from fusions 1 and 2. Rat FLS cells were available for testing hybridomas from all three fusions. TIFF2026515695000009.tif181146TIFF2026515695000010.tif79146

[0226] Example 4: Sequence determination for selected clones To evaluate the clonality between antibodies, selected positive clones were sequenced. RNA was derived from each productive hybridoma, and heavy and light chain cDNA / PCR fragments were generated by 5' SMARTer RACE PCR (TaKaRa) using gene-specific reverse primers constructed in the conserved CH1 region of the respective mouse constant genes for the heavy and light chains. Significant clonal diversity was observed across three fusions for the CDH11-binding antibody. The sequences of the selected antibodies and germline information are provided in Tables 1, 2, and 8.

[0227] (Table 8) Germline information for selected anti-CDH11 antibodies TIFF2026515695000011.tif134146

[0228] Example 5: Effect of anti-CDH11 antibody on FLS infiltration FLS In Vitro Infiltration Test To investigate the functional activity of anti-CDH11 antibodies in cells known to express CDH11, we employed a Matrigel barrier and an in vitro infiltration model. FLS infiltration through Matrigel-coated inserts has been previously described. For example, see Chan A, Akhtar M, Brenner M, Zheng Y, Gulko PS, Symons M. The GTPase Rac Regulates the Proliferation and Invasion of Fibroblast-Like Synoviocytes from Rheumatoid Arthritis Patients. Mol Med. 2007;13(5-6):297-304, which is incorporated in its entirety herein by reference. In this model, FLS infiltration on Matrigel correlates with joint injury. Briefly, we studied FLS infiltration in a Transwell system using Matrigel-coated inserts (BD Biosciences). Cells confluent to 70-80% were collected by trypsin-EDTA digestion and washed with serum-free DMEM. 4 x 10 4Individual (human FLS) or 2 × 10⁻⁶ 4 Cells from rat FLS were resuspended in 500 μl of serum-free DMEM and plated into the upper chamber of a Matrigel-coated insert. Anti-CDH11 antibody was added to the upper chamber at the specified concentration and compared to a commercially available anti-CDH11 antibody (BioLegend, San Diego, CA) and isotype control. The lower chamber was filled with 750 μl of DMEM with 10% FBS, and the plate was incubated at 37°C for 24 hours. After 24 hours, the top surface of the insert was wiped with a cotton swab to remove non-infiltrating cells and the Matrigel layer. The bottom surface of the insert was stained with crystal violet, and the number of cells that had infiltrated through the Matrigel was counted at 100x magnification. All experiments were performed in duplicate. The in vitro invasive properties of FLS measured in this assay have been shown to correlate with radiographic damage in rheumatoid arthritis (RA), accompanied by histological erosive changes in rat arthritis models (Laragione T, Brenner M, Mello A, Symons M, Gulko PS. The arthritis severity locus Cia5d is a novel genetic regulator of the invasive properties of synovial fibroblasts Arthritis Rheum. 2008;58(8):2296-306). (See Tolboom TC, van der Helm-Van Mil AH, Nelissen RG, Breedveld FC, Toes RE, HuizingaTW. Invasiveness of fibroblast-like synoviocytes is an individual patient characteristic associated with the rate of joint destruction in patients with rheumatoid arthritis. Arthritis Rheum. 2005;52(7):1999-2002).

[0229] Efficacy of anti-CDH11 antibody against rat FLS infiltration We used established primary FLS cell lines derived from arthritis-inducing Dark Agouti (DA) rats. These cells are highly invasive and proliferative (doubling in number approximately every 24–48 hours in in vitro culture), making them ideal for initial screening. The in vitro invasiveness of these cell lines correlates with increased disease severity and histological damage in arthritis rats. We investigated the effects of 17 anti-human CDH11 antibodies produced using 3–4 different rat FLS cell lines (Figure 6). Eleven monoclonal anti-human CDH11 antibodies consistently reduced FLS invasiveness, and in the case of five of these antibodies, the effect appeared to be dose-dependent (Figure 6). Two commercially available antibodies, polyclonal goat anti-human CDH11 antibody from R&D Systems (Minneapolis, MN) and IgG1 clone 16G5 antibody from BioLegend (San Diego, CA), were used as positive controls.

[0230] The effect of anti-CDH11 antibodies on rheumatoid arthritis (RA) FLS infiltration. We used established primary FLS cell lines derived from synovial tissue of human patients with rheumatoid arthritis (RA). These cells are highly invasive but do not proliferate as rapidly as rat FLS in vitro. First, we confirmed the consistent expression of CDH11 in 85–91% of RA FLS cells using flow cytometry. The anti-CDH11 antibody IgG1 clone 16G5, commercially available from BioLegend (San Diego, CA), was used for immunostaining and all invasion experiments using human and mouse FLS cells.

[0231] The effects of 27 different anti-human CDH11 monoclonal antibodies on cell infiltration were investigated using three different RA FLS cell lines (Figure 7). Seven of the monoclonal antibodies reduced FLS invasiveness by at least 30% (Figure 7, arrows), and some were dose-dependent.

[0232] Efficacy of anti-CDH11 antibody against mouse FLS infiltration FLS derived from KRN serum-induced arthritis (KSIA) and FLS from collagen antibody-induced arthritis (CAIA) were studied using the Matrigel infiltration model as described above, but 5 × 10⁻⁶ 4 Cells were used. Two monoclonal antibodies (3A10 and 5H1) suppressed KSIA FLS infiltration, two (16A10 and 2F6) showed a variable, dose-independent effect, and one (2A10) was not suppressive (Figure 8A). Five monoclonal antibodies (2A10, 3A10, 5H1, 2F6, and 16A10) reduced CAIA FLS infiltration (Figure 8B). The two antibodies that most consistently reduced mouse FLS infiltration were 3A10 and 5H1, followed by 16A10 and 2F6.

[0233] Example 6: Immunofluorescence staining of mouse FLS with anti-CDH11 antibody FLS cell lines derived from KRN serum-induced arthritis (KSIA) were used for immunofluorescence staining. Of the six antibodies tested, four (16A10, 2A10, 3A10, and 5H1) stained positively compared to the isotype control, while two (2F6 and 2A1) stained negatively (Figure 9).

[0234] Example 7: In vivo efficacy of anti-CDH11 antibody in a KRN serum-induced arthritis (KSIA) model. The ability of monoclonal anti-CDH11 antibodies, 3A10, 2A10, and 10E1, to more significantly reduce arthritis severity scores was tested in vivo using a KSIA model. IgG1 isotype was used as a control. Four to five male C57BL / 6 mice per treatment group received 100 μL of KRN arthritis-inducing serum on days 0 and 2. Starting on day 0, prior to disease onset, mice were treated with either 10 mg / kg of antibody (IgG1 isotype control, 2A10, 3A10, or 10E1) or PBS per day, every other day for a 10-day experimental period (a total of five treatment doses per mouse). Mice were scored every two days. Mice treated with monoclonal 3A10 had significantly lower arthritis severity scores on day 8 compared to the other treatment groups (compared to the PBS group, P=0.03; Figure 10). Figure 11 shows representative images of arthritis mice, demonstrating reduced ankle and foot edema in 3A10 treated mice compared to isotype controls.

[0235] Table 9 outlines the functional screening of anti-CDH11 antibodies.

[0236] (Table 9) Functional screening of anti-CDH11 antibodies. NT = Not tested. *Specificity to CDH11 was not confirmed in experiments using transfected cells for each hybridoma capacity. TIFF2026515695000012.tif152146

[0237] Example 8: Binding affinity of selected anti-CDH11 antibody Based on the binding of antibodies 2A10, 3A10, and 10E1 to transfected human CDH11 Expi293 cells (Figure 12A, Table 10) and mouse CDH11 transfected Expi293 cells (Figure 12B, Table 11), affinity (K) was determined. DThe following was evaluated: KD values ​​were calculated based on nonlinear fitted regression analysis of the combined data (GraphPad Prism). In the third experiment (Figure 12C, Table 12), the binding of antibodies 2A10, 3A10, and 10E1 to transfected human CDH11 Expi293 cells was compared with the binding of two control antibodies, (mAb1) and (mAb2).

[0238] (Table 10) Binding affinity of selected anti-CDH11 antibodies to cellular human CDH11. TIFF2026515695000013.tif24146

[0239] (Table 11) Binding affinity of selected anti-CDH11 antibodies to cellular mouse CDH11. TIFF2026515695000014.tif24146

[0240] (Table 12) Binding affinity of selected anti-CDH11 antibodies to cellular human CDH11. TIFF2026515695000015.tif28146

[0241] Example 9: Domain mapping for selected anti-CHD11 antibodies To determine which CDH11 domain the anti-CDH11 antibody binds to, five cadherin constructs were generated, each lacking one of the five extracellular domains (denoted as cadherin without domain 1, domain 2, domain 3, domain 4, and domain 5). Following the signal peptide and the approximately 30 aa prodomain that is cleaved, the extracellular domains of the cadherins are contiguous, each approximately 110 aa in length. The domains are numbered from the N-terminus (EC1) to the membrane-proximal EC5 (Figure 13). The five domain deletion constructs were fused with GFP at the C-terminus, and binding in transfected Expi293F cells was determined by flow cytometry. The commercially available control 16G5 was used as a control. According to information from the manufacturer of 16G5, this antibody produces antibodies against the EC1 peptide and therefore should be an EC1-binding antibody. In fact, when we evaluated the binding of 16G5, we found that the binding was lost when domains 1 and 2 were removed (Figure 14).

[0242] Anti-CDH11 antibodies 2A10 and 3A10 showed very high binding to transfected human CDH11 but low binding to transfected mouse CDH11. Both antibodies lost binding upon removal of domain 5 (Figure 14A), indicating that these antibodies bind to CDH11 EC5. This domain is also the most different between humans and mice. Anti-CDH11 antibody 10E1 completely lost binding upon removal of domain 4, indicating that this antibody was binding to this domain. Even when adjacent domains 3 and 5 were removed, binding was significantly reduced compared to the full-length construct. Interestingly, when domain 2 was removed, all clones bound better than the full-length construct, indicating that the various domains influence each other. Similarly, when the analysis was extended to a selection of clones with high binding affinity to CDH11, most binding was observed in either domain 4 or 5 (Figure 14B). Serum from immunized mice (Figure 14C) also showed that the polyclonal response was mostly to domains 4 and 5.

[0243] Example 10: Efficacy of anti-CDH11 antibody in a well-established fibrosis model Human hepatic stellate cells (hHSCs) were used as a phenotype for fibrotic diseases. These have been shown to be a major mediator of hepatic fibrosis and are a representative screening method for several fibrotic conditions, including hepatic fibrosis. The commonly tested immortalized hHSC LX2 cell line, the primary hHSC cell line TWNT4 (generated from patients with different liver diseases), and the mouse hepatic stellate cell line JS1 were tested. Cells were cultured for 24 or 48 hours in the presence of different concentrations of anti-CDH11 antibody 3A10 and compared to control isotypes. Anti-CDH11 antibody 3A10 had no effect on the viability (Figure 15A) or proliferation (not shown) of the hHSC cells. Cells were collected at 24 and 48 hours and subjected to triplicate qPCR and quantification of type I collagen (Col1a1) protein in the supernatant. Alk5 inhibitor (Alk5i) was used as a positive control.

[0244] The anti-CDH11 antibody 3A10 significantly reduced the expression of genes involved in fibrosis and the fibrotic phenotype in fibroblasts, hHSCs, and myofibroblasts: type I collagen (Col1a1), actin α2, smooth muscle (Acta2), platelet-derived growth factor receptor β (bPDGFR), matrix metalloproteinase-1 (MMP1), and matrix metalloproteinase-2 (MMP2) (Figure 15B, showing results for LX2, TWNT4, and JS1 cells). The anti-CDH11 antibody 3A10 also significantly reduced Col1a1 protein production by LX-2 and TWNT4, further demonstrating its anti-fibrotic properties (Figures 15C and 15D).

[0245] Anti-CDH11 3A10 significantly reduced the expression of fibrillating genes (Figure 15E), reduced Col1a1 production (Figure 15F), reduced proliferation (Figure 15G), and reduced αSMA protein expression (Figure 15H) in primary hHSCs isolated from patients.

[0246] Example 11: Anti-CDH11 3A10 was not cytotoxic to human precision-cut liver sections (hPCLS). An LDH assay was used to determine cytotoxicity in hPCLS treated with anti-CDH11 3A10. Human precision-cut liver sections (hPCLS) were generated from waste residues of surgically resected human livers. Two different male patients were selected for the hPCLS study as background for liver samples. Normal-looking resection margins surrounding HCC or metastatic areas were selected for hPCLS generation. The ischemic time to PCLS generation after liver resection was 3–4 hours. Resected liver sections were transported to the laboratory in ice-cold Krebs-Henseleit buffer. 8 mm diameter cores were generated using a stainless steel coring instrument, and these liver cores were mounted to specimen holders with solvent-free cyanoacrylate adhesive. The liver cores mounted in specimen holders were mounted in buffer trays immersed in carbogen-saturated ice-cold Krebs-Henseleit buffer supplemented with 25 mM glucose, and liver slices were produced using a VT1200S tissue slicer (Leica Biosystem, IL). Carbogen was continuously supplied to the Krebs-Henseleit buffer between slices. Intact liver slices were collected from the buffer tray and transferred to 6-well tissue culture plates (3-4 slices / well) filled with William's E GlutaMAX medium (Thermo Fisher Scientific, MA) warmed to 37°C, supplemented with 15% hepatocyte medium (Takara Bio Inc.), 25 mM glucose, and 50 μg / ml gentamicin (Thermo Fisher Scientific, MA). To equilibrate the tissue, the slices were pre-incubated for 24 hours on a gentle shaker (10 rpm) in a 37°C incubator supplemented with 5% CO2 and 95% O2. After equilibration in 24 hours of pre-incubation, fresh medium containing IgG1 (1000 ng / ml) control or two different concentrations of 3A10 (100 ng / ml or 1000 ng / ml) was added, and the cells were incubated for an additional 48 hours. At 24 hours after antibody treatment, the old culture medium was replaced with new culture medium containing the respective doses of IgG1 or 3A10 antibody. At the end of drug treatment (at 24 and 48 hours), the culture medium for each condition was collected.To determine cytotoxicity, the luminescence signal of secreted lactate dehydrogenase (LDH) in the culture medium was measured using a Synergy HT (BioTek Instrument Inc., VT) luminometer with the LDH-Glo cytotoxicity assay kit (Promega, WI) as instructed, using purified lactate dehydrogenase derived from rabbit muscle as a standard or positive control.

[0247] As shown in Figure 16, no cytotoxic effects were observed in hPCLS treated with antibody 3A10 for 48 hours.

[0248] Example 12: Anti-CDH11 antibody leads to downregulation of fibrosis genes. LX-2 cells were treated separately for 24 and 48 hours with either 100 ng / ml or 10,000 ng / ml of the hybridoma supernatant containing selected antibodies, respectively. PY102 (10,000 ng / ml) and Alk5i (10 μM) were used in parallel as IgG controls and positive treatment controls. LX-2 cells were harvested 24 and 48 hours after treatment, and total mRNA was purified. RT-qPCR was performed to check the expression of three major fibrinogenic genes: (1) collagen α-1(I) chain (Col1a1, also known as α-1 type I collagen); (2) actin, aortic smooth muscle (Acta2, also known as α-actin-2); and (3) platelet-derived growth factor receptor β (Pdgf-Rβ, also known as platelet-derived growth factor receptor 1 (PDGFR-1)).

[0249] Table 13 shows the downregulation of Col1a1, Acta2, and Pdgf-bR.

[0250] (Table 13) Percentage change in fibrinogenic gene expression in LX-2 cells treated with CDH11 antibody. Downregulation (↓) or upregulation (↑) of fibrinogenic mRNA expression in LX-2 cells treated with the indicated dose of CDH11 antibody compared to the IgG control PY102 (as shown in the bar graph above for mRNA expression). The mean standard error (±SEM) was calculated according to Student's t-test. A p-value < 0.05 was considered statistically significant compared to the PY102 IgG control group. P-values ​​in parentheses are *p<0.05, **p<0.01, ***p<0.001, and ****p<0.0001. TIFF2026515695000016.tif198146TIFF2026515695000017.tif207146

[0251] The selected sequences are provided in Tables 14-19.

[0252] (Table 14) Heavy chain CDR sequences of anti-CDH11 antibodies. SIN = SEQ ID NO. TIFF2026515695000018.tif119146

[0253] (Table 15) Light chain CDR sequences of anti-CDH11 antibodies. SIN = SEQ ID NO. TIFF2026515695000019.tif84146

[0254] (Table 16) Heavy chain variable amino acid sequences of anti-CDH11 antibodies. Antibodies 5A3 and 2E5 have identical sequences. SIN = SEQ ID NO. TIFF2026515695000020.tif235146

[0255] (Table 17) Light chain variable amino acid sequences of anti-CDH11 antibodies. Antibodies 5A3 and 2E5 have identical sequences. SIN = SEQ ID NO. TIFF2026515695000021.tif235146

[0256] (Table 18) Heavy chain variable nucleic acid sequences of anti-CDH11 antibodies. SIN = SEQ ID NO. TIFF2026515695000022.tif138146TIFF2026515695000023.tif222146TIFF2026515695000024.tif218146TIFF2026515695000025.tif138146

[0257] (Table 19) Light chain variable nucleic acid sequences of anti-CDH11 antibodies. SIN = SEQ ID NO. TIFF2026515695000026.tif71146TIFF2026515695000027.tif225146TIFF2026515695000028.tif225146TIFF2026515695000029.tif99146

Claims

1. An anti-cadherin 11 (CDH11) antibody or a fragment thereof that binds to CDH11, The anti-CDH11 antibody or the fragment that binds to CDH11 comprises a heavy chain variable region and a light chain variable region, Each variable region of the heavy chain and light chain includes CDR1, CDR2, and CDR3, (a) CDR1H includes SEQ ID NO: 4, CDR2H includes SEQ ID NO: 13, CDR3H includes SEQ ID NO: 25 or SEQ ID NO: 26, CDR1L includes SEQ ID NO: 38, CDR2L includes SEQ ID NO: 45, and CDR3L includes SEQ ID NO: 53; (b) CDR1H includes SEQ ID NO: 3, CDR2H includes SEQ ID NO: 12, CDR3H includes SEQ ID NO: 23, CDR1L includes SEQ ID NO: 37, CDR2L includes SEQ ID NO: 44, and CDR3L includes SEQ ID NO: 51; (c) CDR1H includes SEQ ID NO: 5, CDR2H includes SEQ ID NO: 14, CDR3H includes SEQ ID NO: 28, CDR1L includes SEQ ID NO: 35, CDR2L includes SEQ ID NO: 43, and CDR3L includes SEQ ID NO: 55; (d) CDR1H includes SEQ ID NO: 6, CDR2H includes SEQ ID NO: 15, CDR3H includes SEQ ID NO: 29, CDR1L includes SEQ ID NO: 40, CDR2L includes SEQ ID NO: 45, and CDR3L includes SEQ ID NO: 56; (e) CDR1H includes SEQ ID NO: 1, CDR2H includes SEQ ID NO: 11, CDR3H includes SEQ ID NO: 20 or SEQ ID NO: 21, CDR1L includes SEQ ID NO: 35, CDR2L includes SEQ ID NO: 43, and CDR3L includes SEQ ID NO: 48 or SEQ ID NO: 49; (f) CDR1H includes SEQ ID NO: 8, CDR2H includes SEQ ID NO: 17, CDR3H includes SEQ ID NO: 32, CDR1L includes SEQ ID NO: 38, CDR2L includes SEQ ID NO: 45, and CDR3L includes SEQ ID NO: 59; (g) CDR1H contains SEQ ID NO: 2, CDR2H contains SEQ ID NO: 11, CDR3H contains SEQ ID NO: 22, CDR1L contains SEQ ID NO: 36, CDR2L contains SEQ ID NO: 43, and CDR3L contains SEQ ID NO: 50; (h) CDR1H includes SEQ ID NO: 3, CDR2H includes SEQ ID NO: 12, CDR3H includes SEQ ID NO: 24, CDR1L includes SEQ ID NO: 35, CDR2L includes SEQ ID NO: 43, and CDR3L includes SEQ ID NO: 52; (i) CDR1H includes SEQ ID NO: 5, CDR2H includes SEQ ID NO: 14, CDR3H includes SEQ ID NO: 27, CDR1L includes SEQ ID NO: 39, CDR2L includes SEQ ID NO: 46, and CDR3L includes SEQ ID NO: 54; (j) CDR1H includes SEQ ID NO: 7, CDR2H includes SEQ ID NO: 16, CDR3H includes SEQ ID NO: 30, CDR1L includes SEQ ID NO: 41, CDR2L includes SEQ ID NO: 44, and CDR3L includes SEQ ID NO: 57; (k) CDR1H includes SEQ ID NO: 7, CDR2H includes SEQ ID NO: 16, CDR3H includes SEQ ID NO: 31, CDR1L includes SEQ ID NO: 35, CDR2L includes SEQ ID NO: 43, and CDR3L includes SEQ ID NO: 58; (l) CDR1H includes SEQ ID NO: 9, CDR2H includes SEQ ID NO: 18, CDR3H includes SEQ ID NO: 33, CDR1L includes SEQ ID NO: 41, CDR2L includes SEQ ID NO: 45, and CDR3L includes SEQ ID NO: 60; or (m) CDR1H includes SEQ ID NO: 10, CDR2H includes SEQ ID NO: 19, CDR3H includes SEQ ID NO: 34, CDR1L includes SEQ ID NO: 42, CDR2L includes SEQ ID NO: 47, and CDR3L includes SEQ ID NO:

61. Anti-cadherin 11 (CDH11) antibody or a fragment of CDH11 that binds to it.

2. (a) CDR1H includes SEQ ID NO: 4, CDR2H includes SEQ ID NO: 13, CDR3H includes SEQ ID NO: 26, CDR1L includes SEQ ID NO: 38, CDR2L includes SEQ ID NO: 45, and CDR3L includes SEQ ID NO: 53; (b) CDR1H includes SEQ ID NO: 3, CDR2H includes SEQ ID NO: 12, CDR3H includes SEQ ID NO: 23, CDR1L includes SEQ ID NO: 37, CDR2L includes SEQ ID NO: 44, and CDR3L includes SEQ ID NO: 51; (c) CDR1H includes SEQ ID NO: 5, CDR2H includes SEQ ID NO: 14, CDR3H includes SEQ ID NO: 28, CDR1L includes SEQ ID NO: 35, CDR2L includes SEQ ID NO: 43, and CDR3L includes SEQ ID NO: 55; (d) CDR1H includes SEQ ID NO: 6, CDR2H includes SEQ ID NO: 15, CDR3H includes SEQ ID NO: 29, CDR1L includes SEQ ID NO: 40, CDR2L includes SEQ ID NO: 45, and CDR3L includes SEQ ID NO: 56; or (e) CDR1H includes SEQ ID NO: 1, CDR2H includes SEQ ID NO: 11, CDR3H includes SEQ ID NO: 21, CDR1L includes SEQ ID NO: 35, CDR2L includes SEQ ID NO: 43, and CDR3L includes SEQ ID NO: 49, The anti-CDH11 antibody or a fragment thereof that binds to CDH11, as described in claim 1.

3. below: (a) A heavy chain variable region containing at least 80% identical sequences to SEQ ID NO: 68, and a light chain variable region containing at least 80% identical sequences to SEQ ID NO: 83; (b) Heavy chain variable region containing at least 80% identical sequences to SEQ ID NO: 65, and light chain variable region containing at least 80% identical sequences to SEQ ID NO: 81; (c) Heavy chain variable region containing at least 80% identical sequences to SEQ ID NO: 70, and light chain variable region containing at least 80% identical sequences to SEQ ID NO: 85; (d) A heavy chain variable region containing at least 80% identical sequences to SEQ ID NO: 71, and a light chain variable region containing at least 80% identical sequences to SEQ ID NO: 86; (e) A heavy chain variable region containing at least 80% identical sequences to SEQ ID NO: 63, and a light chain variable region containing at least 80% identical sequences to SEQ ID NO: 79; (f) A heavy chain variable region containing at least 80% identical sequences to SEQ ID NO: 62, and a light chain variable region containing at least 80% identical sequences to SEQ ID NO: 78; (g) A heavy chain variable region containing at least 80% identical sequences to SEQ ID NO: 75, and a light chain variable region containing at least 80% identical sequences to SEQ ID NO: 90; (h) Heavy chain variable region containing at least 80% identical sequences to SEQ ID NO: 64, and light chain variable region containing at least 80% identical sequences to SEQ ID NO: 80; (i) A heavy chain variable region containing at least 80% identical sequences to SEQ ID NO: 66, and a light chain variable region containing at least 80% identical sequences to SEQ ID NO: 82; (j) A heavy chain variable region containing at least 80% identical sequences to SEQ ID NO: 67, and a light chain variable region containing at least 80% identical sequences to SEQ ID NO: 83; (k) Heavy chain variable region containing at least 80% identical sequences to SEQ ID NO: 69, and light chain variable region containing at least 80% identical sequences to SEQ ID NO: 84; (l) A heavy chain variable region containing at least 80% identical sequences to SEQ ID NO: 72, and a light chain variable region containing at least 80% identical sequences to SEQ ID NO: 87; (m) Heavy chain variable region containing at least 80% identical sequences to SEQ ID NO: 73, and light chain variable region containing at least 80% identical sequences to SEQ ID NO: 88; (n) Heavy chain variable region containing at least 80% identical sequences to SEQ ID NO: 74, and light chain variable region containing at least 80% identical sequences to SEQ ID NO: 89; (o) A heavy chain variable region containing at least 80% identical sequences to SEQ ID NO: 76, and a light chain variable region containing at least 80% identical sequences to SEQ ID NO: 91; (p) A heavy chain variable region containing at least 80% identical sequences to SEQ ID NO: 76, and a light chain variable region containing at least 80% identical sequences to SEQ ID NO: 92; or (q) Heavy chain variable region containing at least 80% identical sequences to SEQ ID NO: 77, and heavy chain variable region containing at least 80% identical sequences to SEQ ID NO: 93 A fragment thereof that binds to the anti-CDH11 antibody or CDH11 according to claim 1 or 2.

4. below: (a) A heavy chain variable region containing at least 90% identical sequences to SEQ ID NO: 68, and a light chain variable region containing at least 90% identical sequences to SEQ ID NO: 83; (b) A heavy chain variable region containing at least 90% identical sequences to SEQ ID NO: 65, and a light chain variable region containing at least 90% identical sequences to SEQ ID NO: 81; (c) Heavy chain variable region containing at least 90% identical sequences to SEQ ID NO: 70, and light chain variable region containing at least 90% identical sequences to SEQ ID NO: 85; (d) A heavy chain variable region containing at least 90% identical sequences to SEQ ID NO: 71, and a light chain variable region containing at least 90% identical sequences to SEQ ID NO: 86; (e) A heavy chain variable region containing at least 90% identical sequences to SEQ ID NO: 63, and a light chain variable region containing at least 90% identical sequences to SEQ ID NO: 79; (f) A heavy chain variable region containing at least 90% identical sequences to SEQ ID NO: 62, and a light chain variable region containing at least 90% identical sequences to SEQ ID NO: 78; (g) A heavy chain variable region containing at least 90% identical sequences to SEQ ID NO: 75, and a light chain variable region containing at least 90% identical sequences to SEQ ID NO: 90; (h) Heavy chain variable region containing at least 90% identical sequences to SEQ ID NO: 64, and light chain variable region containing at least 90% identical sequences to SEQ ID NO: 80; (i) A heavy chain variable region containing at least 90% identical sequences to SEQ ID NO: 66, and a light chain variable region containing at least 90% identical sequences to SEQ ID NO: 82; (j) A heavy chain variable region containing at least 90% identical sequences to SEQ ID NO: 67, and a light chain variable region containing at least 90% identical sequences to SEQ ID NO: 83; (k) Heavy chain variable region containing at least 90% identical sequences to SEQ ID NO: 69, and light chain variable region containing at least 90% identical sequences to SEQ ID NO: 84; (l) A heavy chain variable region containing at least 90% identical sequences to SEQ ID NO: 72, and a light chain variable region containing at least 90% identical sequences to SEQ ID NO: 87; (m) Heavy chain variable region containing at least 90% identical sequences to SEQ ID NO: 73, and light chain variable region containing at least 90% identical sequences to SEQ ID NO: 88; (n) Heavy chain variable region containing at least 90% identical sequences to SEQ ID NO: 74, and light chain variable region containing at least 90% identical sequences to SEQ ID NO: 89; (o) A heavy chain variable region containing at least 90% identical sequences to SEQ ID NO: 76, and a light chain variable region containing at least 90% identical sequences to SEQ ID NO: 91; (p) A heavy chain variable region containing at least 90% identical sequences to SEQ ID NO: 76, and a light chain variable region containing at least 90% identical sequences to SEQ ID NO: 92; or (q) Heavy chain variable region containing at least 90% identical sequences to SEQ ID NO: 77, and light chain variable region containing at least 90% identical sequences to SEQ ID NO: 93 The anti-CDH11 antibody or a fragment thereof that binds to CDH11, as described in claim 3.

5. below: (a) Heavy chain variable region including SEQ ID NO: 68, and light chain variable region including SEQ ID NO: 83; (b) Heavy chain variable region including SEQ ID NO: 65, and light chain variable region including SEQ ID NO: 81; (c) Heavy chain variable region including SEQ ID NO: 70, and light chain variable region including SEQ ID NO: 85; (d) Heavy chain variable region including SEQ ID NO: 71, and light chain variable region including SEQ ID NO: 86; (e) Heavy chain variable region including SEQ ID NO: 63, and light chain variable region including SEQ ID NO: 79; (f) Heavy chain variable region including SEQ ID NO: 62, and light chain variable region including SEQ ID NO: 78; (g) Heavy chain variable region including SEQ ID NO: 75, and light chain variable region including SEQ ID NO: 90; (h) Heavy chain variable region including SEQ ID NO: 64, and light chain variable region including SEQ ID NO: 80; (i) Heavy chain variable region including SEQ ID NO: 66, and light chain variable region including SEQ ID NO: 82; (j) Heavy chain variable region including SEQ ID NO: 67, and light chain variable region including SEQ ID NO: 83; (k) Heavy chain variable region including SEQ ID NO: 69, and light chain variable region including SEQ ID NO: 84; (l) Heavy chain variable region including SEQ ID NO: 72, and light chain variable region including SEQ ID NO: 87; (m) Heavy chain variable region including SEQ ID NO: 73, and light chain variable region including SEQ ID NO: 88; (n) Heavy chain variable region including SEQ ID NO: 74, and light chain variable region including SEQ ID NO: 89; (o) Heavy chain variable region including SEQ ID NO: 76, and light chain variable region including SEQ ID NO: 91; (p) Heavy chain variable region including SEQ ID NO: 76, and light chain variable region including SEQ ID NO: 92; or (q) Heavy chain variable region including SEQ ID NO: 77, and light chain variable region including SEQ ID NO: 93 The anti-CDH11 antibody or a fragment thereof that binds to CDH11, as described in claim 4.

6. The anti-CDH11 antibody or a fragment thereof that binds to CDH11, according to any one of claims 1 to 5, wherein the antibody is a monoclonal antibody.

7. The anti-CDH11 antibody or a fragment thereof that binds to CDH11, according to any one of claims 1 to 6, wherein the anti-CDH11 antibody is a recombinant antibody.

8. A fragment thereof that binds to an anti-CDH11 antibody or CDH11 according to any one of claims 1 to 7, comprising a human framework region or a modified human framework region.

9. A fragment thereof that binds to an anti-CDH11 antibody or CDH11 according to any one of claims 1 to 8, comprising a human constant region or a modified human constant region.

10. The fragments that bind to the aforementioned CDH11 are scFv, Fv, Fab', Fab, F(ab') 2 A fragment that is coupled to the CDH11 according to any one of claims 1 to 9, or a diamond body.

11. A deglycosylated anti-CDH11 antibody or a fragment thereof that binds to CDH11, according to any one of claims 1 to 10.

12. A fragment thereof that binds to an anti-CDH11 antibody or CDH11 according to any one of claims 1 to 11, conjugated with one or more of a cytotoxin, a fluorescent label, and an agent for image analysis.

13. A nucleic acid molecule encoding an anti-CDH11 antibody or a fragment thereof that binds to CDH11, as described in any one of claims 1 to 11.

14. below: (a) SEQ ID NO: 100 and / or SEQ ID NO: 117; (b) SEQ ID NO: 97 and / or SEQ ID NO: 114; (c) SEQ ID NO: 102 and / or SEQ ID NO: 119; (d) SEQ ID NO: 103 and / or SEQ ID NO: 120; (e) SEQ ID NO: 95 and / or SEQ ID NO: 112; (f) SEQ ID NO: 94 and / or SEQ ID NO: 111; (g) SEQ ID NO: 96 and / or SEQ ID NO: 113; (h) SEQ ID NO: 98 and / or SEQ ID NO: 115; (i) SEQ ID NO: 99 and / or SEQ ID NO: 116; (j) SEQ ID NO: 101 and / or SEQ ID NO: 118; (k) SEQ ID NO: 104 and / or SEQ ID NO: 121; (l) SEQ ID NO: 105 and / or SEQ ID NO: 122; (m) SEQ ID NO: 106 and / or SEQ ID NO: 123; (n) SEQ ID NO: 107 and / or SEQ ID NO: 124; (o) SEQ ID NO: 108 and / or SEQ ID NO: 125; (p) SEQ ID NO: 109 and / or SEQ ID NO: 126; or (q) SEQ ID NO: 110 and / or SEQ ID NO: 127 A nucleic acid molecule according to claim 13, comprising:

15. A vector or set of vectors comprising the nucleic acid molecule described in claim 13 or 14.

16. An isolated host cell comprising a nucleic acid molecule according to claim 13 or 14, or a vector or set of vectors according to claim 15.

17. The step of culturing host cells under conditions in which an anti-CDH11 antibody or a fragment thereof that binds to CDH11 is produced by the host cells described in claim 16. A method for producing an anti-CDH11 antibody or a fragment thereof that binds to CDH11, including the above.

18. A pharmaceutical composition comprising an anti-CDH11 antibody or a fragment thereof that binds to CDH11, as described in any one of claims 1 to 12, and a pharmaceutically acceptable excipient.

19. A method for inhibiting intercellular CDH11-CDH11 interactions, The step of bringing the cells into contact with an anti-CDH11 antibody or a fragment thereof that binds to CDH11, as described in any one of claims 1 to 12, or the pharmaceutical composition described in claim 18. Methods that include...

20. A method for inhibiting intercellular CDH11-CDH11 interactions in subjects where this is necessary, A step of administering to a subject an anti-CDH11 antibody or a fragment thereof that binds to CDH11 according to any one of claims 1 to 12, or a pharmaceutical composition according to claim 18. Methods that include...

21. The method according to claim 19 or 20, comprising the step of inhibiting the CDH11-CDH11 interaction between hepatic stellate cells (HSCs).

22. A method for treating fibrosis in a person who needs it, A step of administering to a subject an anti-CDH11 antibody or a fragment thereof that binds to CDH11 according to any one of claims 1 to 12, or a pharmaceutical composition according to claim 18. Methods that include...

23. The method according to claim 22, wherein the fibrosis is hepatic fibrosis, pulmonary fibrosis, renal fibrosis, cardiac fibrosis, myelofibrosis, pancreatic fibrosis, cutaneous fibrosis, scleroderma, intestinal fibrosis, post-myocardial infarction fibrosis, post-replacement fibrosis, perivascular fibrosis, or articular fibrosis.

24. The method according to claim 23, wherein the fibrosis is hepatic fibrosis.

25. The method according to claim 24, wherein the subject has an infection caused by a hepatitis virus, non-alcoholic fatty liver disease (NASH), hereditary metabolic disorder, cholestatic disorder, or immunological disorder.

26. The method according to claim 24, wherein the liver fibrosis is associated with increased alcohol intake, excessive vitamin A intake, or drug toxicity.

27. The method according to claim 24, wherein the liver fibrosis is related to an autoimmune disease of the liver or bile duct, liver graft rejection, or a genetic disorder of the liver or bile duct.

28. The method according to claim 27, wherein the autoimmune disease of the liver or bile duct is autoimmune hepatitis, primary biliary cholangitis, or primary sclerosing cholangitis.

29. The method according to claim 27, wherein the genetic disorder of the liver is Wilson's disease or hemochromatosis, or the genetic disorder of the bile duct is biliary atresia.

30. The method according to claim 23, wherein the fibrosis is intestinal fibrosis.

31. The method according to claim 30, wherein the subject has inflammatory bowel disease (IBD), Crohn's disease, or colitis.

32. The method according to claim 23, wherein the pulmonary fibrosis is induced by idiopathic pulmonary fibrosis or toxic injury.

33. The method according to claim 20 or 22, wherein the subject has scleroderma or systemic scleroderma.

34. The method according to claim 20 or 22, wherein the subject has keloid scarring.

35. A method for treating rheumatoid arthritis in a person who needs it, A step of administering to a subject an anti-CDH11 antibody or a fragment thereof that binds to CDH11 according to any one of claims 1 to 12, or a pharmaceutical composition according to claim 18. Methods that include...

36. The method according to any one of claims 20 to 35, wherein the subject is a human.

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