Antibody to integrin alpha 11 beta 1

Abstract

To provide a novel function blocking antibody to type I collagen receptor integrin alpha 11 beta 1 (α11β1).SOLUTION: Provided are: an anti-integrin alpha 11 beta 1 (α11β1) antibody including (1) a heavy chain including heavy chain complementarity determining region 1 (CDRH1), CDRH2 and CDRH3, and (2) a light chain including light chain complementarity determining region 1 (CDRL1), CDRL2 and CDRL3, where each of the CDRH1 to CDRH3 and CDRL1 to CDRL3 includes an amino acid sequence consisting of a specific sequence; or an antigen-binding fragment thereof; and a method of fabricating and using the antibody. In some embodiments, the anti-α11β1 antibody or the antigen-binding fragment thereof is a monoclonal antibody or an antigen-binding fragment thereof. Also provided is a use of the antibody for treating fibrous disorders and / or cancer.SELECTED DRAWING: None

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application was filed on December 20, 2019, each of which is incorporated herein in its entirety. U.S. Provisional Application No. 62 / 951,723 filed February 28, 2020 No. 62 / 983,155 filed on July 21, 2020; This application claims priority to U.S. Provisional Application No. 63 / 054,717. [Background technology]

[0002] Fibrosis occurs in many tissues in the body, typically as a result of inflammation or tissue injury. Increased production of extracellular matrix leads to organ failure. Fibrosis-related diseases account for 100% of all deaths in industrialized countries. It accounts for approximately 45% of all cases (Wynn, TA, 2008, J Pathol. 214:199-210). One of the most common diseases is systemic sclerosis (SSc). SSc is a complex autoimmune disease that progresses over a chronic period. SSc has a progressive course and is highly variable among patients. It is characterized by inflammation, vascular dysfunction, and fibrosis. Fibrosis of the skin and internal organs leads to irreversible scarring and ultimately organ failure. Currently, there are no approved drugs with disease-modifying potential. There is no effective treatment. Summary of the Invention

[0003] The present disclosure relates to the type I collagen receptor integrin alpha 11 beta 1 (α11β1). The present disclosure also provides novel function-blocking antibodies against fibrotic disorders and / or cancer. The use of such antibodies for therapy is provided.

[0004] In one aspect, the present disclosure provides a method for the preparation of a nucleic acid sequence comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 103-435. In another aspect, the present invention provides an anti-α11β1 antibody or antigen-binding fragment thereof, comprising a sequence. The figures show SEQ ID NOs: 103 to 207, 209, 211, 213, 216, 218, 220, 2 23, 225, 228, 233, 234, 236, 240, 241, 245, 247, 2 53, 255, 257, 259, 261, 265, 267, 269, 271, 275, 2 77, 279, 281, 283, 287, 289, 291, 293, 296, 300, 3 04, 306, 308, 310, 312, 314, 316, 318, 320, 322, 3 24, 325, 327, 329, 334, 336, 338, 340, 342, 344, 3 48, 351, 353, 355, 358, 360, 361, 364, 366, 368, 3 69, 374, 376, 377, 379, 380, 381, 383, 384, 385, 3 87, 389, 392, 393, 396, 398, 400, 402, 405, 408, 4 Anti-α11β antibody containing a CDR sequence included in any one of 11, or 413 to 435 In another aspect, the present disclosure provides an antibody or antigen-binding fragment thereof, comprising SEQ ID NO: 103. CDR1, CDR2, and Anti-α11β1 antibodies or antigen-binding fragments thereof, including CDR3, are provided. In some embodiments, the anti-α11β1 antibody or antigen-binding fragment thereof is selected from the group consisting of SEQ ID NOs: 103-114, The amino acid sequence is selected from the group consisting of 207-311 and 312-435. In some embodiments, the anti-α11β1 antibody or antigen-binding fragment thereof is SEQ ID NO: 10 3-114, 207, 209, 211, 213, 216, 218, 220, 223, 22 5, 228, 233, 234, 236, 240, 241, 245, 247, 253, 25 5, 257, 259, 261, 265, 267, 269, 271, 275, 277, 27 9, 281, 283, 287, 289, 291, 293, 296, 300, 304, 30 6, 308, 310, 312, 314, 316, 318, 320, 322, 324, 32 5, 327, 329, 334, 336, 338, 340, 342, 344, 348, 35 1, 353, 355, 358, 360, 361, 364, 366, 368, 369, 37 4, 376, 377, 379, 380, 381, 383, 384, 385, 387, 38 9, 392, 393, 396, 398, 400, 402, 405, 408, 411, and In some embodiments, the CDR sequence is included in any one of 413 to 435. The anti-α11β1 antibody or antigen-binding fragment thereof is selected from the group consisting of SEQ ID NOs: 103 to 114 and 41. 3 to 434. In embodiments, the anti-α11β1 antibody or antigen-binding fragment thereof is selected from the group consisting of SEQ ID NOs: 103-114. or CDR1, CDR2, and CDR3 encompassed by any one of 413 to 434 Includes:

[0005] In some embodiments, the anti-α11β1 antibody or antigen-binding fragment thereof is a monoclonal antibody. In some embodiments, the antibody is an anti-α11β1 antibody or an antigen-binding fragment thereof. or an antigen-binding fragment thereof is a humanized antibody or an antigen-binding fragment thereof. In embodiments, the anti-α11β1 antibody or antigen-binding fragment thereof is isolated from human α11β1-expressing cells. In some embodiments, the anti-α11β1 antibody reduces the interaction between α11β1 and collagen in the The α11β1 antibody or antigen-binding fragment thereof may be any of the antibodies or antigen-binding fragments thereof described herein. Competes with sexual fragments.

[0006] In another aspect, the disclosure provides a nucleic acid encoding an antibody or antigen-binding fragment thereof described herein. In some embodiments, the nucleic acid sequence is selected from the group consisting of SEQ ID NOs: 1 to 5. 102.

[0007] In another aspect, the disclosure provides a vector comprising a nucleic acid described herein.

[0008] In another aspect, the disclosure provides a method for producing a nucleic acid comprising the nucleic acid described herein or the vector described herein. A host cell containing the vector is provided.

[0009] In another aspect, the present disclosure provides a method for producing an antibody or antigen-binding fragment thereof comprising administering the antibody or antigen-binding fragment thereof under conditions suitable for expression. producing the antibody or antigen-binding fragment thereof, comprising culturing the host cell described herein. A method is provided.

[0010] In another aspect, the present disclosure provides a method for treating a subject having or at risk for a fibrotic disorder. A method for administering a therapeutically effective amount of an antibody or antigen-binding fragment thereof described herein to a subject In some embodiments, the method comprises administering to a subject in need thereof: Fibrotic disorders include idiopathic pulmonary fibrosis (IPF), chronic kidney disease, diabetic cardiomyopathy, and primary sclerosing pulmonary disease (PSD). Cholangitis (PSC), primary biliary cirrhosis (PBC), non-alcoholic fatty liver disease (NA FLD / NASH), Crohn's disease, ulcerative colitis, or systemic sclerosis.

[0011] In another aspect, the present disclosure provides a method for treating a subject having or at risk of cancer. A method comprising administering a therapeutically effective amount of an antibody or antigen-binding fragment thereof described herein to a subject. In some embodiments, the method comprises administering to a subject in need thereof a method for treating cancer. head and neck squamous cell carcinoma, pancreatic ductal adenocarcinoma, non-small cell lung cancer, adrenocortical carcinoma, acute myeloid leukemia, bladder cancer Bladder urothelial carcinoma, invasive breast cancer, cervical squamous cell carcinoma, bile duct cancer, colorectal adenocarcinoma, diffuse large cell carcinoma Alveolar B-cell lymphoma, esophageal adenocarcinoma, glioblastoma multiforme, hepatocellular carcinoma, lung adenocarcinoma, squamous lung epithelial carcinoma, cutaneous melanoma, mesothelioma, ovarian serous cystadenocarcinoma, pheochromocytoma and paraganglioma, prostate Adenocarcinoma, sarcoma, gastric adenocarcinoma, testicular germ cell tumor, thymoma, thyroid cancer, endometrial cancer of the uterine corpus, uterine Among carcinosarcoma, uveal melanoma, renal clear cell carcinoma, renal carcinosarcoma, and renal papillary cell carcinoma One or more of: [Brief explanation of the drawings]

[0012] The teachings described herein, when read in conjunction with the accompanying drawings, include various exemplary embodiments as follows: The present invention will be more fully understood from the following description of the preferred embodiments. are for illustrative purposes only and are not intended to limit the scope of the present teachings in any way. I want to be understood. [Figure 1] Figure 1 shows the structure of integrins. The panel shows the structure of a collagen-binding integrin, and three different conformations of integrins can exist on the cell surface. [Figure 2A] FIG. 2A shows a chart depicting ELISA analysis of exemplary mouse monoclonal antibodies for binding to human α11β1. [Figure 2B] FIG. 2B shows a chart depicting an exemplary ELISA analysis of binding of mouse monoclonal antibodies to mouse α11β1. [Figure 3A] FIG. 3A shows a graph depicting an ELISA analysis of binding of an exemplary rat monoclonal antibody to the human α11β1 I domain. [Figure 3B] FIG. 3B shows a graph depicting an ELISA analysis of binding of exemplary mouse monoclonal antibodies to the human α11β1I domain. [Figure 4A] FIG. 4A shows a graph depicting a FACS analysis of binding of an exemplary rat monoclonal antibody to human α11β1-expressing CHO-K1 cells. [Figure 4B] FIG. 4B shows a graph depicting a FACS analysis of binding of exemplary mouse monoclonal antibodies to human α11β1-expressing CHO-K1 cells. [Figure 5] FIG. 5 shows a graph depicting a FACS analysis of binding of exemplary murine monoclonal antibodies to human lung fibroblasts (HPF) and myofibroblasts (MF). [Figure 6A] FIG. 6A shows a graph depicting the ability of exemplary rat monoclonal antibodies to inhibit adhesion of CHO-K1 cells expressing human α11 to rat tail type I collagen. [Figure 6B] FIG. 6B shows a graph depicting the ability of exemplary rabbit monoclonal antibodies to inhibit adhesion of CHO-K1 cells expressing human α11 to rat tail type I collagen. [Figure 6C] FIG. 6C shows a graph depicting the ability of exemplary mouse monoclonal antibodies to inhibit adhesion of CHO-K1 cells expressing human α11 to rat tail type I collagen. [Figure 7A] FIG. 7A shows a graph depicting the ability of exemplary rat monoclonal antibodies to inhibit fibroblast-to-myofibroblast transition (FMT) as measured by percent inhibition of αSMA upregulation. [Figure 7B]FIG. 7B shows a graph depicting the ability of exemplary rabbit monoclonal antibodies to inhibit fibroblast to myofibroblast transition (FMT) as measured by percent inhibition of αSMA upregulation. [Figure 7C] FIG. 7C shows a graph depicting the ability of exemplary murine monoclonal antibodies to inhibit fibroblast to myofibroblast transition (FMT), as measured by percent inhibition of αSMA upregulation. [Figure 8] FIG. 8 shows a graph depicting the ability of exemplary monoclonal antibodies to inhibit CHO-K1 human α11-mediated rat tail collagen type I gel contraction. [Figure 9] FIG. 9 shows a graph depicting the affinity of exemplary monoclonal antibodies to human α11β1 by surface plasmon resonance (SPR). [Figure 10A] 10A and 10B show graphs depicting the affinity of exemplary monoclonal antibodies to human α11β1 by surface plasmon resonance (SPR). [Figure 10B] 10A and 10B show graphs depicting the affinity of exemplary monoclonal antibodies to human α11β1 by surface plasmon resonance (SPR). [Figure 11A] Figures 11A and 11B show graphs depicting the ability of selected rabbit, rat, mouse, and human monoclonal antibodies to bind to α11β1 expressed on the surface of CHO cells. [Figure 11B] Figures 11A and 11B show graphs depicting the ability of selected rabbit, rat, mouse, and human monoclonal antibodies to bind to α11β1 expressed on the surface of CHO cells. [Figure 12] FIG. 12 shows a graph depicting a FACS analysis of binding of exemplary monoclonal antibodies to human lung fibroblasts (HPF) and myofibroblasts (MF). [Figure 13] FIG. 13 shows a graph and table depicting FACS analysis of binding of exemplary monoclonal antibodies to human myofibroblasts (MF). [Figure 14]FIG. 14 shows a graph depicting the binding ability of selected monoclonal antibodies to α11β1 expressed on the surface of CHO cells. [Figure 15A] 15A and 15B show graphs depicting the ability of exemplary monoclonal antibodies to inhibit adhesion of CHO cells expressing human alpha 11 to rat tail type I collagen. [Figure 15B] 15A and 15B show graphs depicting the ability of exemplary monoclonal antibodies to inhibit adhesion of CHO cells expressing human alpha 11 to rat tail type I collagen. [Figure 16] FIG. 16 shows a graph depicting the effect of exemplary monoclonal antibodies on xenograft growth in SCID mice. [Figure 17A] Figures 17A, 17B, and 17C depict the effect of exemplary monoclonal antibodies on soluble pro-fibrogenic markers in Precision-Cut Liver Slices (PCLS). [Figure 17B] Figures 17A, 17B, and 17C depict the effect of exemplary monoclonal antibodies on soluble pro-fibrogenic markers in Precision-Cut Liver Slices (PCLS). [Figure 17C] Figures 17A, 17B, and 17C depict the effect of exemplary monoclonal antibodies on soluble pro-fibrogenic markers in Precision-Cut Liver Slices (PCLS). DETAILED DESCRIPTION OF THE INVENTION

[0013] Detailed Description The present disclosure is based, in part, on the discovery of novel antibodies that selectively bind to α11β1. The disclosure also provides nucleic acids encoding said antibodies, as well as methods for treating fibrosis and fibrotic components. and methods of use in the treatment of diseases including

[0014] Fibrosis and Disease Fibrosis occurs in many tissues in the body, typically as a result of inflammation or tissue injury. Increased production of extracellular matrix leads to organ failure. Fibrosis-related diseases account for 100% of all deaths in industrialized countries. It accounts for approximately 45% of all cases (Wynn, TA, 2008, J Pathol. 214:199-210). One of the most common diseases is systemic sclerosis (SSc). SSc is a complex autoimmune disease that progresses over a chronic period. SSc has a progressive course and is highly variable among patients. It is characterized by inflammation, vascular dysfunction, and fibrosis. Fibrosis of the skin and internal organs leads to irreversible scarring and ultimately organ failure. Currently, there are no approved drugs with disease-modifying potential. There is no effective treatment.

[0015] Responsible for the production of extracellular matrix (ECM) for tissue repair (and in fibrosis) The cells that cause fibrosis are a special type of fibroblast called myofibroblasts (MF). Although the mechanisms have been widely studied, this complex process is still not fully understood. To focus on the most important drivers of fibrosis, we have compiled published patient-derived data sets. The data set (SSc patient data and normal controls) was analyzed using a novel data analysis method obtained in-house. This analysis revealed that type I collagen-binding integrin alpha 11-based α11β1 was identified as one of the most important targets modulating fibrosis .

[0016] To date, there are no true disease-modifying therapies for fibrosis. The two currently approved therapies, nintedanib and pirfenidone, have insufficient efficacy and are associated with poor disease progression. There are no approved therapies for systemic sclerosis (SSc) to date, and no disease-modifying treatments are currently available. In some embodiments, the fibrotic disorder is idiopathic pulmonary fibrosis (IPF), chronic kidney disease, diabetes, or the like. Diabetic cardiomyopathy, primary sclerosing cholangitis (PSC), primary biliary cirrhosis (PBC), non-alcoholic liver cirrhosis fatty liver disease (NAFLD / NASH), Crohn's disease, ulcerative colitis, or In some embodiments, the fibrosis is or comprises systemic sclerosis (SSc). The disorders include atrial fibrosis, endomyocardial fibrosis, arthrofibrosis, mediastinal fibrosis, myelofibrosis, and progressive myelofibrosis. Progressive massive fibrosis, retroperitoneal fibrosis, or skeletal muscle fibrosis Being or containing them.

[0017] One of the clinical features of the tumor microenvironment is the interaction between the tumor and stroma, which is primarily This depends on various integrins that interact with ECM components as well as growth factors. Such interactions may affect tumor survival, progression, and ultimately metastasis. β1 has been reported to be overexpressed in cancer-associated fibroblasts (CAFs) in metastatic tumors. Its expression correlates with malignant tumors in patients. It is overexpressed in the stroma of most head and neck squamous cell carcinomas (HNSCC) and is an alpha-smooth muscle There was a positive correlation with cutin expression (Parajuli et al., J. Oral Pathol. Med. 46:267-275 ( Integrin α11 is also overexpressed by CAFs in the stroma of pancreatic ductal adenocarcinoma (PDAC). (Schnittert et al., FASEB J. 33:6609-6621 (2019)). Overexpression of integrin α11β1 in the stroma is a key factor in the progression of non-small cell lung cancer (NSCLC). It is associated with tumor growth and metastatic potential, and ITGA11 (integrin alpha-1 in humans) High expression of the gene encoding IL-1 is associated with a prolonged recurrence-free survival in all NSCLC patients. In the same study, overexpression of α11 in lung cancer cell lines was associated with decreased migration and invasion. It has been shown to increase the risk of developing glaucoma (Ando et al., Cancer Sci. 111:200-208 (2020)).

[0018] Integrin Integrins are a large family of type I transmembrane heterodimeric glycoprotein receptors. They act as the primary receptors for cell adhesion. Modulates signaling pathways that regulate cell adhesion, migration, proliferation, differentiation, and apoptosis It consists of 18 α subunits and 8 β subunits. These combine to form 24 integrin heterodimers. Each integrin receptor consists of two non-covalently linked subunits, α and β. Integrins α1β1, α2β1, α10β1, and α11β1 are major The α and β subunits are large modular extracellular domains. It is a transmembrane protein with a single transmembrane helix and a short cytoplasmic region. Mediates cytoskeletal interactions. The extracellular domains of integrins are generally large, approximately 80–1 It is a 50 kDa structure. The extracellular domain consists of a headpiece connected to two legs. (See Figure 1 for the structure of collagen-binding integrins.) 1). Collagen-binding integrins function as ligand-binding sites. The αI domain binds to divalent metal cations (Mg2+) and binds to ligands. The conserved "metal ion-dependent adhesion site" (MI) plays an important role in ATP binding. Contains DAS).

[0019] Integrins can exist in three different conformations: 1) resting; A low affinity state (curved conformation, Figure 1, Panel A) containing the ligand binding site. 2) an elongated, intermediate-affinity state with the headpiece facing the membrane; The headpiece remains "closed" while the ligament is extended (Figure 1, panel B), and ) an extended, high-affinity state in which the integrin is fully activated and readily binds to its ligand. The complexity of different integrin states allows for the creation of alloproteins that inhibit integrin function. Both telic and ligand-blocking approaches are possible. As indicated by the asterisk in Figure 1, One allosteric way to block integrin function is to block the integrin-dependent elongation pathway. A monoclonal antibody that prevents the ATP from reaching the fully extended conformation from a restricted intermediate conformation. Another allosteric option is to generate a curved / inactive conformation. It binds to integrins in the endothelial cell structure and prevents them from elongating into either of the other two states. A non-allosteric way to inhibit integrin function is to bind to the I domain. The ligand binding site prevents integrins from binding to collagen. Direct binding to runs the risk of generating a recombinant activator of integrin function.

[0020] As cell surface receptors, integrins sense the stiffness of the surrounding matrix and mediate the function of connective tissue This induces cells to further generate and remodel cells, which perpetuates the fibrotic phenotype. Many integrins are overexpressed in fibrosis, but which alpha subunits are overexpressed? It is unclear whether the α11β1 integrin is sufficient for the development of fibrosis. is specifically expressed in a subset of fibroblasts and myofibroblasts (i.e., peripheral scar-producing cells) Recent literature has shown that α11β1 is expressed in cardiac tissue, liver, lung, and kidney. Strong evidence has been presented that IFN-γ is one of the major drivers of the fibrotic phenotype (Romaine, A. et. al. Overexpression of integrin alpha 11 induces cardiac fibros is in mice. Acta Physiol Feb 2018, 222(2); Bansal, R. et.al. Integrin alpha 11 i n the regulation of the myofibroblast phenotype: implications for fibrotic disea ses. Exp Mol Med. 2017 Nov 17:49(11)). Blocking α11β1 function improves muscle fiber function. Inhibition of fibroblast differentiation and extracellular matrix deposition (i.e., key events in scar formation) Blocking α11β1 function may also fundamentally alter the fibrotic microenvironment. It is possible to modify the progression of all diseases with fibrotic components by altering the fibrotic component. This may provide a mechanism for localized, injury-specific attenuation of fibrosis.

[0021] In some embodiments, the anti-α11β1 antibodies or antigen-binding fragments thereof of the present disclosure are It reduces the interaction between α11β1 and collagen in α11β1-expressing cells. In some embodiments, the interaction between α11β1 and collagen in human α11β1-expressing cells is The reduced affinity of α11β1 allows it to interact with α11β1 in a resting, low-affinity state (curved conformation). In some embodiments, the antibody comprises an anti-α11β1 antibody or antigen-binding fragment thereof. The reduced interaction of α11β1 with collagen in α11β1-expressing cells was observed after elongation. Anti-α11β1 antibodies or antigen binding thereof that interact with α11β1 in an intermediate affinity state In some embodiments, the α11β1 and α11β1 in human α11β1-expressing cells are Reduced interaction with collagen interacts with α11β1 in its extended, high-affinity state. The present invention includes an anti-α11β1 antibody or an antigen-binding fragment thereof.

[0022] antibody The term "antibody" is used herein in its broadest sense and includes, but is not limited to: Monoclonal antibodies, polyclonal antibodies, multispecific antibodies (e.g., bispecific antibodies) and / or antibody fragments (preferably fragments that exhibit the desired antigen-binding activity). The antibodies described herein include immunoglobulins, heavy chain antibodies, light chain antibodies, and the like. , LRR-based antibodies, or other protein scaffolds with antibody-like properties, as well as those described in the art. Other immunological binding moieties known in the art, such as Fab, Fab', Fab'2, Fa b2, Fab3, F(ab')2, Fd, Fv, Feb, scFv, SMIP, antibody, da Ear body, Tria body, Tetra body, Mini body, Maxi body, Tandubu, DV D, BiTe, TandAb, etc., or any combination thereof. The subunit structures and three-dimensional configuration of antibodies are known in the art.

[0023] A "monoclonal antibody" or "mAb" is obtained from a substantially homogeneous population of antibodies. The antibodies in the population are potential variant antibodies (e.g., For example, it may contain naturally occurring mutations or mutations that arise during the manufacture of a monoclonal antibody preparation. are identical and / or bind to the same epitope, except that Different antigens are typically present in trace amounts. Polyclonal antibody preparations contain antibodies that are identical to each other, whereas monoclonal antibody preparations contain antibodies that are identical to each other. A clonal antibody is directed against a single determinant on an antigen.

[0024] An "antigen-binding fragment" is an intact antibody that binds to the antigen to which the intact antibody binds. The antigen-binding fragment of an antibody specifically binds to an antigen to form a complex. Naturally occurring, enzymatically derived, synthetic, or genetically engineered polypeptides or glycoproteins. Exemplary antibody fragments include, but are not limited to, Fv , Fab, Fab', Fab'-SH, F(ab')2; diabody; linear antibody; single chain antibody molecules (e.g., scFv or VHH or VH or VL domains only); and In some embodiments, the antibodies described herein include multispecific antibodies formed from antibody fragments. The antigen-binding fragment of the antibody described in is an scFv. The fragments may be monospecific or multispecific (e.g., bispecific). A multispecific antigen-binding fragment of an antibody may contain at least two different variable domains. Each variable domain may be directed to a different antigen or to a different epitope of the same antigen. It is capable of specifically binding to

[0025] A "multispecific antibody" is a small antibody that recognizes and specifically binds to at least two different antigens. A "bispecific antibody" refers to an antibody that contains at least two different antigen-binding domains. A type of antibody that recognizes and specifically binds to at least two different antigens. Refers to an antibody that contains an antigen-binding domain.

[0026] "Different antigens" are different and / or distinct proteins, polypeptides, or molecules. and may refer to different and / or distinct epitopes, although these epitopes The fragments may be contained within a single protein, polypeptide, or other molecule.

[0027] The term "epitope" refers to a specific region within the variable region of an antibody molecule known as a paratope. A single antigen may have multiple epitopes. Thus, different antibodies can bind to different regions of an antigen and bind to different organisms. The term "epitope" also refers to an antigen to which B cells and / or T cells respond. It also refers to the region of an antigen to which an antibody binds. Functional epitopes are generally defined as structural epitopes. An epitope is a subset of a given protein that contains residues that directly contribute to the affinity of the interaction. It may also be a conformational epitope, ie, composed of non-linear amino acids. In certain embodiments, an epitope is an amino acid, a sugar side chain, a phosphoryl group, or a sulfonyl group. The determinant may be a chemically active surface group of a molecule, such as a phenyl group, and may be a specific In embodiments, the antibody may have particular three-dimensional structural characteristics and / or particular charge characteristics.

[0028] As used herein, "selective binding," "selectively bind," and "specific binding" or "specifically binds" refers to, with respect to an antigen-binding moiety and an antigen target, This refers to the fact that a molecule binds preferentially to its antigen target and does not bind to entities that are not antigen targets. Some degree of non-specific binding may occur between the binding moiety and non-targets. In this study, the binding between the antigen-binding moiety and the antigen target was compared to the binding between the antigen-binding moiety and the non-target. More than 2 times, more than 5 times, more than 10 times, or more than 100 times In some embodiments, the antigen-binding moiety selectively binds to an antigen target. The moiety has a binding affinity of approximately 10 -5 Less than M, about 10 -6 Less than M, about 10 -7 Less than M, about 10 -8 Less than M or about 10 -9 If the α-amino acid is less than M, it will selectively bind to the antigen target.

[0029] In some embodiments, selective binding to the same or overlapping epitopes Comparable antibodies or fragments thereof often cross-compete for binding to the antigen. Thus, in some embodiments, the present disclosure provides an exemplary antibody or fragment thereof disclosed herein. In some embodiments, "cross-competing" antibodies or fragments thereof are provided. "Compete," "compete," "cross-compete," or "compete" refers to the ability of an antibody or fragment thereof to bind to a target This means that the two molecules compete for the same epitope or binding site. , the reference antibody or fragment thereof prevents or inhibits specific binding of the test antibody or fragment thereof. It can be determined by assay and vice versa. Competitive binding assays are used to determine whether a test molecule competes with a reference molecule for binding. Examples of assays that can be used include solid phase direct or indirect Radioimmunoassay (RIA), solid-phase direct or indirect enzyme immunoassay (EIA), Sandwich competition assays (e.g., Stahli et al. (1983) Methods in Enzymology 9:2 42-253), solid-phase direct biotin-avidin EIA (e.g., Kirkland et al., (1986 ) J. Immunol. 137:3614-9), solid-phase direct labeling assay, solid-phase direct labeling sandwich Assay, Luminex (Jia et al. "A novel method of Multiplexed Competitive Antibody Binning for the characterization of monoclonal antibodies” J. Immunol ocular Methods (2004) 288, 91-98), and surface plasmon resonance (Song et al. “Epit ope Mapping of Ibalizumab, a Humanized Anti-CD4 Monoclonal Antibody with Anti-HI V-1 Activity in Infected Patients” J. Virol. (2010) 84, 6935-42). Typically, when a competing antibody or fragment thereof is present in excess, it will compete with the reference antibody or fragment thereof. The fragments bind to the common antigen by at least 50%, 55%, 60%, 65%, 70%, or In some cases, binding is inhibited by at least 80%, 85%, 90%, 95% , 96%, 97%, 98%, 99% or more inhibited.

[0030] Antibodies consist of four polypeptide chains, e.g., two heavy (H) chains and two light (L) chains. In some embodiments, the light chain is a lambda light chain. In some embodiments, the light chain is a kappa light chain. A heavy chain comprises a heavy chain variable domain and a heavy chain variable domain. The heavy chain constant domain can include a CH1, hinge, CH2, C The light chain comprises a light chain variable domain and an H3 region and optionally a CH4 region. The light chain constant domain may comprise a CL and a light chain constant domain. can be done.

[0031] The heavy chain variable domain of the heavy chain and the light chain variable domain of the light chain are typically The complementarity-determining regions (CDs) are interspersed with more conserved regions called FRs. These heavy chain variable domains and The light chain variable domains are arranged in the following order from amino terminus to carboxyl terminus: Three Cs arranged as R1, CDR1, FR2, CDR2, FR3, CDR3, FR4 DR and four framework regions, one or more of which The CDRs of the heavy chain may be modified as described herein. The CDRs of the light chain are called "CDRH1," "CDRH2," and "CDRH3," and the CDRs of the light chain are called "CDRL These are referred to as "CDRL1," "CDRL2," and "CDRL3."

[0032] There are five major classes of antibodies: IgA, IgD, IgE, IgG, and IgM Some of these are subclasses (isotypes), e.g., IgG1, IgG2 Immunoglobulins are further classified into IgG, IgG3, IgG4, IgA1, and IgA2. The heavy chain constant domains corresponding to the different classes of α, δ, ε, γ, and μ, respectively, are Called.

[0033] Exemplary Antibodies The present disclosure provides antibodies that can include the various heavy and light chains described herein. In some embodiments, the antibody comprises two heavy chains and a light chain. Thus, the present disclosure provides a method for the production of at least one heavy chain and / or light chain disclosed herein, at least one heavy and / or light chain framework domain disclosed in the specification, at least one heavy and / or light chain CDR domain as disclosed herein; and and / or an antibody comprising any heavy and / or light chain constant domain disclosed herein. Includes.

[0034] In some embodiments, the antibodies disclosed herein are homodimeric monoclonal antibodies. In some embodiments, the antibodies disclosed herein are heterodimeric antibodies. In some embodiments, the antibody is, for example, a typical antibody or diabody, Tria Body, Tetra Body, Mini Body, Maxi Body, Tandubu, DVD, BiTe , scFv, TandAb scFv, Fab, Fab2, Fab3, F(ab')2, or any combination thereof.

[0035] The present disclosure relates to, inter alia, anti-integrin alpha11 beta1 (α11β1) antibodies, In some embodiments, the α11β1 antibody or its antigen-binding fragments are provided. The antigen-binding fragment comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 103 to 435. In some embodiments, the anti-α11β1 antibody or antigen-binding fragment thereof is selected from the group consisting of SEQ ID NO: 103-207, 209, 211, 213, 216, 218, 220, 223, 225, 228, 233, 234, 236, 240, 241, 245, 247, 253, 255, 257, 259, 261, 265, 267, 269, 271, 275, 277, 279, 281, 283, 287, 289, 291, 293, 296, 300, 304, 306, 308, 310, 312, 314, 316, 318, 320, 322, 324, 325, 327, 329, 334, 336, 338, 340, 342, 344, 348, 351, 353, 355, 358, 360, 361, 364, 366, 368, 369, 374, 376, 377, 379, 380, 381, 383, 384, 385, 387, 389, 392, 393, 396, 398, 400, 402, 405, 408, 411, or 4 In some embodiments, the CDR sequence is comprised within any one of SEQ ID NOs: 13 to 435. The anti-α11β1 antibody or antigen-binding fragment thereof is selected from the group consisting of SEQ ID NOs: 103 to 206 and 413 to 416. 435. In some embodiments, the anti-α11β1 antibody or antigen-binding fragment thereof is selected from the group consisting of SEQ ID NOs: 103-104. 14, 207-311, and 312-435. In some embodiments, the anti-α11β1 antibody or antigen-binding fragment thereof comprises SEQ ID NO: No. 103~114, 207, 209, 211, 213, 216, 218, 220, 223 , 225, 228, 233, 234, 236, 240, 241, 245, 247, 253 , 255, 257, 259, 261, 265, 267, 269, 271, 275, 277 , 279, 281, 283, 287, 289, 291, 293, 296, 300, 304 , 306, 308, 310, 312, 314, 316, 318, 320, 322, 324 , 325, 327, 329, 334, 336, 338, 340, 342, 344, 348 , 351, 353, 355, 358, 360, 361, 364, 366, 368, 369 , 374, 376, 377, 379, 380, 381, 383, 384, 385, 387 , 389, 392, 393, 396, 398, 400, 402, 405, 408, 411 or 413 to 435. In the present invention, the anti-α11β1 antibody or antigen-binding fragment thereof is selected from the group consisting of SEQ ID NOs: 103 to 114 and 4. 13 to 434. In one embodiment, the anti-α11β1 antibody or antigen-binding fragment thereof is selected from the group consisting of SEQ ID NOs: 103-11 4, or 413 to 434. In some embodiments, the anti-α11β1 antibody or antigen-binding fragment thereof comprises: In some embodiments, the antibody is an anti-α1 monoclonal antibody or an antigen-binding fragment thereof. The 1β1 antibody or antigen-binding fragment thereof is a humanized antibody or antigen-binding fragment thereof. In some embodiments, the anti-α11β1 antibody or antigen-binding fragment thereof is human α11β 1-expressing cells. In one embodiment, the present disclosure provides a method for the preparation of a nucleic acid sequence comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 103 to 435. and an anti-α11β1 antibody or antigen-binding fragment thereof that competes with the antibody or antigen-binding fragment thereof. In some embodiments, the present disclosure provides fragments of the group consisting of SEQ ID NOs: 103-435. an anti-α11 antibody or an antigen-binding fragment thereof, which competes with the antibody or antigen-binding fragment thereof, comprising an amino acid sequence selected from β1 antibodies or antigen-binding fragments thereof are provided.

[0036] In some embodiments, the present disclosure provides a method for the preparation of a medicament comprising administering to a subject a heavy chain as provided herein and a medicament comprising administering to a subject a heavy chain as provided herein. The present invention provides an anti-α11β1 antibody or antigen-binding fragment thereof, comprising a light chain that is In embodiments, the present disclosure provides a heavy chain variable domain as provided herein and a The present invention provides an anti-α11β1 antibody or an antigen-binding fragment thereof, comprising a light chain variable region that is In some embodiments, the present disclosure provides specific combinations of heavy and light chain variable domains. Anti-α11β1 antibodies or antigen-binding fragments thereof are provided, including combinations thereof. In this embodiment, the anti-α11β1 antibody or antigen-binding fragment thereof comprises a heavy chain selected from Table 1. It comprises a combination of a single chain variable domain and a light chain variable domain.

[0037] [Table 1]

[0038] In some embodiments, the present disclosure provides an antibody against an anti-α11β1 antibody or antigen-binding fragment thereof. 1 to 30 (e.g., 1, 2, 3, 4, 5, 10, or more) additions, deletions, or an anti-α11β1 antibody or an antigen-binding fragment thereof, comprising a deletion or substitution; 1 antibody is selected from the group consisting of SEQ ID NOs: 103 to 158, 413, 414, and 421 to 434 The antibody or fragment may contain a selected amino acid sequence, e.g., a sequence that selectively binds to α11β1. In some embodiments, an anti-α11β1 antibody or antigen-binding fragment thereof is provided, wherein the antibody is The present disclosure provides an anti-α11β1 antibody or an antigen-binding fragment thereof, which has 1 to 30 additions, deletions, or or an anti-α11β1 antibody or antigen-binding fragment thereof containing a substitution, The antibody is selected from the group consisting of SEQ ID NOs: 103 to 114, 413, 414, and 421 to 434. and a fragment thereof, which comprises an amino acid sequence selected from the group consisting of: In some embodiments, the present invention provides an anti-α11β1 antibody or antigen-binding fragment thereof. The disclosure provides SEQ ID NOs: 103-158, 413, 414, and 421-434. At least 80%, 85%, 90%, 91%, 92% for the selected amino acid sequence; 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity an anti-α11 β1 antibody or fragment thereof, which comprises an amino acid sequence, e.g., an antibody or fragment thereof, which selectively binds to α11 β1; In some embodiments, the present disclosure provides a β1 antibody or antigen-binding fragment thereof. Selected from the group consisting of column numbers 103 to 114, 413, 414 and 421 to 434 At least 80%, 85%, 90%, 91%, 92%, 93%, 9 containing an amino acid sequence with 4%, 95%, 96%, 97%, 98% or 99% identity For example, an anti-α11β1 antibody or fragment thereof, wherein the antibody or fragment selectively binds to α11β1. The antigen-binding fragments thereof are provided.

[0039] In some embodiments, the present disclosure provides an antibody against an anti-α11β1 antibody or antigen-binding fragment thereof. 1 to 90 (e.g., 1 to 50, e.g., at least 1, 2, 3, 4, 5, 6, 7, anti-α11β1 antibodies or their derivatives containing additions, deletions, or substitutions of 8, 9, or 10 or more amino acids; an antigen-binding fragment of an anti-α11β1 antibody or an antigen-binding fragment thereof, The amino acid sequence is selected from the group consisting of 9 to 206 and 415 to 420, for example an anti-α11β1 antibody or its antigen-binding fragment, wherein the antibody or fragment selectively binds to α11β1; In some embodiments, the present disclosure provides synthetic fragments of SEQ ID NOs: 159-206 and and 415-420, and 5%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or contains an amino acid sequence with 99% identity, e.g., an antibody or fragment thereof may be The present invention provides an anti-α11β1 antibody or antigen-binding fragment thereof that selectively binds to the α11β1 antibody.

[0040] In some embodiments, the present disclosure provides an antibody or fragment thereof that selectively binds to α11β1. fragment, and one or more C shown in the list of exemplary sequences provided herein For example, in some embodiments, antibodies or fragments thereof are provided that comprise DR sequences. or fragments thereof, comprising one or more CDRs of SEQ ID NOs: 103 to 114. In some embodiments, the present disclosure provides an antibody or fragment thereof that selectively binds to α11β1. Therefore, at least 95% of one or more CDRs of SEQ ID NOs: 103 to 114 96%, 97%, 98%, or 99% identical to an antibody or In some embodiments, the antibody or fragment is selected from the group consisting of SEQ ID NOs: 103-104, 104-106, 106-108, 107-109, 108-109, 109-200, 110-111, 111-112, 112-113, 113-114, 114-115, 115-11 At least 90%, 91%, 92%, 93%, 94%, 95% for one of 14 , 96%, 97%, 98%, or 99% identity to the amino acid sequence of the antibody. , comprising one or more CDRs set forth in one of SEQ ID NOs: 103-114. For example, the antibody or fragment may have a sequence similar to that of SEQ ID NO: 103, but not identical .... , 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity The antibody comprises one or more CDRs (e.g., CDRs 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 3 For example, one, two, or three CDRs.

[0041] In some embodiments, the present disclosure provides an antibody or fragment thereof that selectively binds to α11β1. fragment, and one or more C shown in the list of exemplary sequences provided herein For example, in some embodiments, antibodies or fragments thereof are provided that comprise DR sequences. or fragments thereof are SEQ ID NOs: 103 to 207, 209, 211, 213, 216, 218, 220, 223, 225, 228, 233, 234, 236, 240, 241, 245, 247, 253, 255, 257, 259, 261, 265, 267, 269, 271, 275, 277, 279, 281, 283, 287, 289, 291, 293, 296, 300, 304, 306, 308, 310, 312, 314, 316, 318, 320, 322, 324, 325, 327, 329, 334, 336, 338, 340, 342, 344, 348, 351, 353, 355, 358, 360, 361, 364, 366, 368, 369, 374, 376, 377, 379, 380, 381, 383, 384, 385, 387, 389, 392, 393, 396, 398, 400, 402, 405, 408, 411, or one or more CDRs of 413 to 435. In some embodiments, the present disclosure provides an antibody or fragment thereof that selectively binds to α11β1. SEQ ID NOs: 103 to 207, 209, 211, 213, 216, 218, 220, 22 3, 225, 228, 233, 234, 236, 240, 241, 245, 247, 25 3, 255, 257, 259, 261, 265, 267, 269, 271, 275, 27 7, 279, 281, 283, 287, 289, 291, 293, 296, 300, 30 4, 306, 308, 310, 312, 314, 316, 318, 320, 322, 32 4, 325, 327, 329, 334, 336, 338, 340, 342, 344, 34 8, 351, 353, 355, 358, 360, 361, 364, 366, 368, 36 9, 374, 376, 377, 379, 380, 381, 383, 384, 385, 38 7, 389, 392, 393, 396, 398, 400, 402, 405, 408, 41 1, or at least 95% for one or more CDRs 413-435; an antibody or its derivatives containing an amino acid sequence that is 96%, 97%, 98%, or 99% identical to In some embodiments, the antibody or fragment is selected from the group consisting of SEQ ID NOs: 103-207. , 209, 211, 213, 216, 218, 220, 223, 225, 228, 233 , 234, 236, 240, 241, 245, 247, 253, 255, 257, 259 , 261, 265, 267, 269, 271, 275, 277, 279, 281, 283 , 287, 289, 291, 293, 296, 300, 304, 306, 308, 310 , 312, 314, 316, 318, 320, 322, 324, 325, 327, 329 , 334, 336, 338, 340, 342, 344, 348, 351, 353, 355 , 358, 360, 361, 364, 366, 368, 369, 374, 376, 377 , 379, 380, 381, 383, 384, 385, 387, 389, 392, 393 , 396, 398, 400, 402, 405, 408, 411, or 413-435 At least 90%, 91%, 92%, 93%, 94%, 95%, 96% for one of them %, 97%, 98%, or 99% identity to the amino acid sequence of the sequence Numbers 103 to 207, 209, 211, 213, 216, 218, 220, 223, 22 5, 228, 233, 234, 236, 240, 241, 245, 247, 253, 25 5, 257, 259, 261, 265, 267, 269, 271, 275, 277, 27 9, 281, 283, 287, 289, 291, 293, 296, 300, 304, 30 6, 308, 310, 312, 314, 316, 318, 320, 322, 324, 32 5, 327, 329, 334, 336, 338, 340, 342, 344, 348, 35 1, 353, 355, 358, 360, 361, 364, 366, 368, 369, 37 4, 376, 377, 379, 380, 381, 383, 384, 385, 387, 38 9, 392, 393, 396, 398, 400, 402, 405, 408, 411, and comprises one or more CDRs set forth in any one of 413 to 435. For example, an antibody or the fragment is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, 101%, 102%, 103%, 104%, 105%, 106%, 107%, 108%, 109%, 110%, 111, 112%, 113%, 114%, 115%, Amino acid sequences with 4%, 95%, 96%, 97%, 98%, or 99% identity and the antibody comprises one or more CDRs (e.g., 1, 2, or or three CDRs).

[0042] The present disclosure provides, inter alia, methods for producing anti-α11β1 antibodies or antigen-binding fragments thereof. Methods for generating antibodies are known in the art. In some embodiments, The disclosure provides a method for producing antibodies or antigen-binding fragments thereof from SEQ ID NOs: 1-102 under conditions suitable for expression of the antibodies or antigen-binding fragments thereof. and culturing a host cell containing nucleic acid comprising a nucleic acid sequence selected from the group consisting of: or a method for producing an antigen-binding fragment thereof.

[0043] Exemplary Nucleotide Sequences The present disclosure provides one or more heavy chains, heavy chain variable domains, heavy chain framework regions, heavy chain CDR, heavy chain constant domain, light chain, light chain variable domain, light chain framework region, light chain C DR, light chain constant domain, or other immunoglobulin-like sequence disclosed herein, antibody or a nucleotide sequence encoding the binding molecule. Such a nucleotide sequence may be present in a vector. Such nucleotides can be administered to cells, e.g., cells of a subject in need of treatment, or cells that produce antibodies. It may be present in the genome of a cell for producing the gene, for example, a mammalian cell for producing an antibody.

[0044] In some embodiments, the present disclosure provides a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 103-206. a nucleic acid comprising a nucleic acid sequence encoding an antibody or an antigen-binding fragment thereof comprising an amino acid sequence In some embodiments, the present disclosure provides a method for the treatment of a leukemia, comprising administering to a patient a method for the treatment of a leukemia, comprising administering to a patient a method for the treatment of a leukemia, a leukemia, or ... a leukemia, or a leukemia, a leukemia, or a leukemia, a A nucleic acid sequence encoding an antibody or antigen-binding fragment thereof comprising a selected amino acid sequence. In some embodiments, the present disclosure provides nucleic acids comprising the group consisting of SEQ ID NOs: 1-102. In some embodiments, the present disclosure provides a nucleic acid comprising a nucleic acid sequence selected from the group consisting of: A vector containing a nucleic acid comprising a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 1 to 102 is provided. In some embodiments, the present disclosure provides a method for the preparation of a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 1-102. In some embodiments, the present disclosure provides a host cell comprising a nucleic acid comprising the nucleic acid sequence. A vector containing a nucleic acid comprising a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 1 to 102 is provided. do.

[0045] In some embodiments, the present disclosure provides a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 1-102. At least 80%, 85%, 90%, 91%, 92%, 93%, 94% of the acid sequence Nucleic acids containing nucleic acid sequences with 95%, 96%, 97%, 98%, or 99% sequence identity Provides acid.

[0046] Measurement of antibody interaction with α11β1 The binding properties of the antibodies described herein to α11β1 can be determined by methods known in the art, for example, by the method described below. Methods below: BIACORE analysis, enzyme-linked immunosorbent assay (ELISA), x-ray crystallography The activity of the antibody can be determined by one of structural analysis, sequence analysis, and scanning mutagenesis. The binding interaction between the antibody and α11β1 can be analyzed using surface plasmon resonance (SPR). SPR or Biomolecular Interaction Analysis (BIA) can be used to determine how interacting substances are detected. BIA chips detect biospecific interactions in real time without labeling. A change in mass at the binding surface (indicating a binding event) changes the refractive index of light near the surface. Changes in refractive index produce detectable signals, indicative of real-time reactions between biomolecules The method using SPR is described, for example, in U.S. Pat. No. 5,641,640. Specification;Raether (1988) Surface Plasmons Springer Verlag;Sjolander and Urbaniczk y (1991) Anal. Chem. 63:2338-2345; Szabo et al. (1995) Curr. Opin. Struct. Biol. 5:699-705, and BIAcore International AB (Sweden) , Uppsala). KinExA (registered trademark) available from e Instruments (Boise, Idaho) The Kinetic Exclusion Assay™ assay can also be used. can.

[0047] Information from SPR was used to determine the equilibrium dissociation constant (K) for antibody binding to α11β1. D ) and K on and K. off provides accurate and quantitative measurements of kinetic parameters, including Such data can be used to compare different molecules. Information from SPR can also be used to develop structure-activity relationships (SAR). Identifying variant amino acids at a given position that correlate with binding parameters, e.g., high affinity It is possible.

[0048] In certain embodiments, the antibodies described herein exhibit high affinity binding to α11β1. In various embodiments, the K of the antibodies described herein for α11β1 is D is about 1 0 -4 , 10 -5 , 10 -6 , 10 -7 , 10 -8 , 10 -9 , 10 -10 , 10 -11 , 10 -12 , 10 -13 , 10 -14 , or 10 -15 In certain cases, In this case, the K D The range is 0.001 to 1 nM. For example, 0.001nM, 0.005nM, 0.01nM, 0.05nM, 0.1n M, 0.5 nM, or 1 nM.

[0049] Treatment method In some embodiments, one or more anti-α11β1 antibodies described herein are one or more disorders as described herein, for example, one or more fibrotic disorders and / or or one or more cancers. The method comprises administering a therapeutically effective amount of an antibody or antigen-binding fragment thereof described herein to a subject. In some embodiments, the fibrotic disorder is an idiopathic fibrotic disorder. Pulmonary fibrosis (IPF), chronic kidney disease, diabetic cardiomyopathy, primary sclerosing cholangitis (PSC), Primary biliary cirrhosis (PBC), nonalcoholic fatty liver disease (NAFLD / NASH), Crohn's disease, ulcerative colitis, or systemic sclerosis. In some embodiments, the fibrotic disorder is atrial fibrosis, endomyocardial fibrosis, arthrofibrosis, mediastinal fibrosis, or the like. fibrosis, myelofibrosis, progressive massive fibrosis, retroperitoneal fibrosis, or skeletal muscle fibrosis; or including them.

[0050] In some embodiments, one or more anti-α11β1 antibodies described herein are For example, one or more of the following cancers: head and neck squamous cell carcinoma, pancreatic ductal adenocarcinoma, non-small cell lung cancer, Renal cortical carcinoma, acute myeloid leukemia, urothelial carcinoma of the bladder, invasive breast cancer, squamous cell carcinoma of the cervix, bile duct Cancer, colorectal adenocarcinoma, diffuse large B-cell lymphoma, esophageal adenocarcinoma, glioblastoma multiforme , liver hepatocellular carcinoma, lung adenocarcinoma, lung squamous cell carcinoma, cutaneous melanoma, mesothelioma, ovarian serous cystadenocarcinoma, brown Chromocytoma and paraganglioma, prostate adenocarcinoma, sarcoma, gastric adenocarcinoma, testicular germ cell tumor, thymoma, thyroid Adenocarcinoma, endometrial cancer of the uterine corpus, uterine carcinosarcoma, uveal melanoma, renal clear cell carcinoma, renal carcinosarcoma and in methods for treating renal papillary cell carcinoma.

[0051] Combination therapy In some embodiments, the anti-α11β1 antibodies described herein may be administered with one or more additional The present invention is administered in combination with an additional therapeutic agent, such as a chemotherapeutic or oncolytic agent. As used herein, "combination therapy" means two or more different overlapping drug regimens. This refers to a situation where a subject is exposed to both drugs simultaneously. When administered in combination, the two or more different agents may be administered simultaneously or separately. The term "co-administration of two or more drugs in the same dosage form, in separate dosage forms, and in separate dosage forms" refers to the simultaneous administration of two or more drugs in the same dosage form, in separate dosage forms, and in separate dosage forms. i.e., two or more drugs are formulated together in the same dosage form and administered simultaneously. Alternatively, two or more agents may be administered simultaneously, where the agents are in separate formulations. In another alternative, after the first agent is administered, one or more additional agents may be administered. In separate administration protocols, two or more drugs may be administered minutes apart or may be administered several hours apart or several days apart.

[0052] As used herein, the term "chemotherapeutic agent" or "oncolytic therapeutic agent" (e.g., an anti- Cancer drugs (e.g., anti-cancer therapies, e.g., immune cell therapy) are, for example, for use in the treatment of one or more diseases, disorders or conditions associated with excessive cell proliferation one or more pro-apoptotic agents, including agents utilized and / or recommended for , cytostatic and / or cytotoxic, and / or hormonal agents, In some embodiments, the chemotherapeutic agent and / or tumor inhibitor is a Tumor regression therapeutic agents include platinum compounds (e.g., cisplatin, carboplatin, and oxalate). platin), alkylating agents (e.g., cyclophosphamide, ifosfamide, chloramphenicol, Bucil, nitrogen mustard, thiotepa, melphalan, busulfan, procal vasodilator, streptozocin, temozolomide, dacarbazine, and bendamustine), anti- Antitumor antibiotics (e.g., daunorubicin, doxorubicin, idarubicin, epirubicin) , mitoxantrone, bleomycin, mitomycin C, plicamycin, and dac tinomycin), taxanes (e.g., paclitaxel and docetaxel), antimetabolites ( For example, 5-fluorouracil, cytarabine, premetrexed, thiouracil guanine, floxuridine, capecitabine, and methotrexate), nucleosides analogs (e.g., fludarabine, clofarabine, cladribine, pentostatin, and and nelarabine), topoisomerase inhibitors (e.g., topotecan and irinotecan), Methylating agents (e.g., azacitidine and decitabine), proteosome inhibitors (e.g., bovine serum albumin), lutezomib), epipodophyllotoxins (e.g., etoposide and teniposide), DN A synthesis inhibitors (e.g., hydroxyurea), vinca alkaloids (e.g., bicristine , vindesine, vinorelbine, and vinblastine), tyrosine kinase inhibitors (e.g., (e.g., imatinib, dasatinib, nilotinib, sorafenib, and sunitinib), nitro ureas (e.g., carmustine, fotemustine, and lomustine), hexamethylmethacrylate lamins, mitotane, angiogenesis inhibitors (e.g., thalidomide and lenalidomide), steroids (e.g., prednisone, dexamethasone, and prednisolone), hormones drugs (e.g., tamoxifen, raloxifene, leuprolide, bicalatomid, granice) thoron, and flutamide), aromatase inhibitors (e.g., letrozole and anastomoses) Trozole), arsenic trioxide, tretinoin, nonselective cyclooxygenase inhibitors (e.g. Nonsteroidal anti-inflammatory drugs, salicylates, aspirin, piroxicam, ibuprofen indomethacin, naprosyn, diclofenac, tolmetin, ketoprofen, nab Meton, and Oxaprozin), selective cyclooxygenase-2 (COX-2) inhibitors The agent may be or may include a steroid agent, a steroid drug, or any combination thereof.

[0053] In certain embodiments, chemotherapeutic agents for anti-cancer treatment and / or oncolytic treatment The agent may be a biological agent, e.g., a tumor-infiltrating lymphocyte, a CAR T cell, an antibody, an antigen, a therapeutic agent, Vaccines (e.g., antigens produced by a patient's own tumor cells or a specific tumor) other substances, such as steroids), immunomodulators (e.g., cytokines, e.g., immunomodulators, or biological response modifiers), checkpoint inhibitors, or other immunological agents In certain embodiments, immunological agents include immunoglobulins, immunostimulants (e.g., For example, bacterial vaccines, colony-stimulating factors, interferons, interleukins, therapeutic vaccines, vaccine combinations, viral vaccines) and / or immunosuppressants (e.g., calcitonin, These include: steroid inhibitors, steroid agonists ... In certain embodiments, the hormonal agent includes an agent for antiandrogen therapy (e.g., ketoconazole, Nazol, abiraterone, TAK-700, TOK-OO1, bicalutamide, nilutamide , flutamide, enzalutamide, ARN-509).

[0054] Additional chemotherapeutic and / or oncolytic therapeutic agents include immune checkpoint therapeutics ( For example, pembrolizumab, nivolumab, ipilimumab, atezolizumab, avelumab, durvalumab, tremelimumab, or cemiplimab), other monoclonal antibodies (e.g. For example, rituximab, cetuximab, panetumumab, tositumomab, trastuzumab, and Lemtuzumab, gemtuzumab, ozogamicin, bevacizumab, catumaxomab, denosumab Mab, obinutuzumab, ofatumumab, ramucirumab, pertuzumab, nimotuzumab, Lambrolizumab, pidilizumab, siltuximab, BMS-936559, RG744 6 / MPDL3280A, MEDI4736), antibody-drug conjugates (e.g., Brentsky Cimab vedotin; (ADCETRIS®, Seattle Genetic s); ado-trastuzumab emtansine (KADCYLA®, Roche ); gemtuzumab ozogamicin (Wyeth); CMC-544; SAR3419; CDX-011; PSMA-ADC; BT-062; and IMGN901 (e.g., Sa ssoon et al., Methods Mol. Biol. 1045:1-27 (2013); Bouchard et al., Bioorganic M ed. Chem. Lett. 24: 5357-5363 (2014)), or any combination thereof. do.

[0055] In some embodiments, the combined administration of an anti-α11β1 antibody and an additional therapeutic agent enhances the anti-α11β1 in cancer compared with that achieved by either the 1β1 antibody or the additional therapeutic agent alone The difference between the combined effect and the effect of each drug alone can be statistically significant. In some embodiments, the combined effect can be a synergistic effect. The combined administration of the anti-α11β1 antibody and the additional therapeutic agent may be performed using standard administration regimens, e.g., The administration of additional therapeutic agents compared to the approved dosing regimen for the other therapeutic agents The amount may be reduced, administered less frequently, and / or administered less frequently.

[0056] In some embodiments, the methods of treatment described herein are useful in treating a medical condition when other treatments have failed. or in subjects who have had little success with treatment by other means. Additionally, the methods of treatment described herein may include one or more additional treatments for the medical condition. For example, the method can be performed in combination with an anti-α11β antibody described herein. a cancer regimen, prior to, substantially simultaneously with, or following administration of one or more antibodies or compositions thereof; For example, administering non-myeloablative chemotherapy, surgery, hormone therapy, and / or radiation may include:

[0057] Formulation and Administration In various embodiments, the antibodies described herein can be incorporated into pharmaceutical compositions. Such pharmaceutical compositions can be used, for example, for the prevention and / or treatment of diseases, such as fibrotic disorders. The pharmaceutical compositions may be prepared by methods known to those skilled in the art (e.g., Remington's P harmaceutical Sciences, 17th edition, ed. Alfonso R. Gennaro, Mack Publishing Co The formulation may be carried out by methods such as those described in the company, Easton, Pa. (1985). Cut.

[0058] In some embodiments, the pharmaceutical composition comprises a pharmaceutically acceptable carrier or excipient. Examples of pharmaceutically acceptable carriers include, but are not limited to, However, any and all physiologically compatible solvents, dispersion media, coatings, antimicrobial agents and Antifungal agents, isotonic and absorption delaying agents, etc. The compositions of the present invention may contain pharmaceutically acceptable carriers. These may include salts which are formed by addition of the compounds of formula (I) or (II), such as acid addition salts or base addition salts.

[0059] In some embodiments, compositions comprising the antibodies described herein, e.g., sterile injectable formulations, The agent can be formulated in accordance with conventional pharmaceutical practice using water for injection as a vehicle. For example, physiological saline or glucose as well as D-sorbitol, D-mannose, Isotonic solutions containing D-mannitol and other supplements such as sodium chloride are optionally Suitable solubilizing agents, for example alcohols such as ethanol and / or propylene glycol polyalcohols such as alcohol or polyethylene glycol, and / or polysorbates Injection in combination with a nonionic surfactant such as PEG-80 or HCO-50 It can be used as an aqueous solution for injection.

[0060] As disclosed herein, pharmaceutical compositions may be in any form known in the art. Such forms include, for example, liquid, semi-solid and solid dosage forms, e.g., liquid solutions ( For example, injection and infusion solutions), dispersions or suspensions, tablets, pills, powders, liposomal preparations, These include suppositories and injectables.

[0061] The selection or use of any particular form will depend, in part, on the intended mode of administration and therapeutic application. For example, compositions including compositions intended for systemic or local delivery The composition may be in the form of an injectable or infusible solution. To be administered by injection (e.g., intravenous, subcutaneous, intraperitoneal, or intramuscular injection) As used herein, parenteral administration refers to administration by injection. refers to modes of administration other than enteral and topical administration by, including but not limited to, intravenous, Intranasal, intraocular, intrapulmonary, intramuscular, intraarterial, intrathecal, intravesical, intraorbital, intracardiac, intradermal, intrapulmonary, abdominal Intracavitary, transtracheal, subcutaneous, subcuticular, intra-articular, subcapsular, subarachnoid, intraspinal, epidural, intracerebral, cranial These include intravenous, intracarotid and intrasternal injections and infusions.

[0062] The route of administration can be parenteral, e.g., by injection, intranasal, pulmonary, or transdermal. Administration can be by intravenous, intramuscular, intraperitoneal, or subcutaneous injection. This can be done systemically or locally.

[0063] In some embodiments, the pharmaceutical compositions of the present invention are in the form of a solution, a microemulsion, Prepared as dispersions, liposomes, or other ordered structures suitable for stable storage at high concentrations Sterile injectable solutions can be prepared by combining the compositions described herein in the required amount in an appropriate solvent. The mixture is optionally combined with one or a combination of the above listed ingredients, followed by Generally, dispersions can be prepared by sterilizing with a filter using the methods described herein. The compositions are then dissolved in a sterile vehicle containing the basic dispersion medium and the required other ingredients from those enumerated above. In the case of a sterile powder for preparing a sterile injection solution, For example, the preparation method includes mixing a powder of the composition described herein with any additional desired ingredients (see below). These include vacuum drying and freeze drying, which are obtained by obtaining the above-mentioned solutions (reference) from previously sterile-filtered solutions. The proper fluidity of the dispersion can be achieved, for example, by the use of a coating such as lecithin. This can be achieved by maintaining the required particle size when necessary and by using surfactants. Prolonged absorption of the injectable compositions can be achieved by using an agent delaying absorption, for example, monostearin This can be achieved by including an acid salt, and gelatin in the composition.

[0064] Pharmaceutical compositions include sterile solutions or suspensions in water or another pharmaceutically acceptable liquid. It can be administered parenterally in the form of an injection. For example, the pharmaceutical composition can be administered in the form of a pharmaceutically acceptable biosimilar. Vehicles or media, such as sterile water and saline, vegetable oils, emulsifiers, suspending agents, surfactants, and therapeutic molecules, such as activators, stabilizers, flavoring excipients, diluents, vehicles, preservatives, binders, etc. and subsequently blended in a unit dosage form as required by generally accepted pharmaceutical practice. The amount of active ingredient contained in the pharmaceutical preparation can be determined by the specified The amount is such that an appropriate dosage within the range specified is provided. Non-limiting examples of oily liquids include: Contains corn and soybean oils, with benzyl benzoate or benzyl alcohol as a solubilizer. Other items that can be included are phosphate buffer or acetate. Buffers such as sodium buffer, sedatives such as procaine hydrochloride, benzyl alcohol or The formulated injections contain stabilizers such as phenol and antioxidants. Can be packaged in a pull.

[0065] In various embodiments, subcutaneous administration is by syringe, pre-filled syringe, auto-injector (e.g. disposable or reusable), pen-type injectors, patch-type injectors, wearables Syringe, ambulatory syringe infusion pump with subcutaneous infusion set, or antibody for subcutaneous injection This can be achieved by devices such as other devices for combining with drugs.

[0066] The injection system of the present disclosure is similar to that described in U.S. Pat. No. 5,308,341. A delivery pen can be used as follows: Commonly used pen devices are well known in the art. It may contain at least one syringe needle (e.g., a 31-gauge needle approximately 5-8 mm in length). , typically pre-filled with one or more therapeutic unit doses of therapeutic solution, Some drug delivery pens are useful for delivering drugs to a subject as quickly and painlessly as possible. , including a vial holder that can hold a vial of therapeutic or other drug. The device may be a completely mechanical device or may be combined with electronic circuitry to allow the user to The dose of medicine to be injected into the syringe can also be precisely set and / or displayed. For example, See U.S. Patent No. 6,192,891. In some embodiments, the pen The needles in the device are disposable, and the kit includes one or more disposable replacement needles. A pen device suitable for delivering any one of the presently characterized compositions may be, for example, See, for example, U.S. Patent Nos. 6,211,629 and 6,232,797, the disclosures of each of which are incorporated herein by reference in their entireties. Specification No. 77,099; Specification No. 6,200,296; Specification No. 6,146,361 Microneedle-based pen devices are also described in, for example, the disclosures thereof. No. 7,556,615, which is incorporated herein by reference in its entirety. Manufactured by Scandinavian Health Ltd. Precision Pen Injector (PPI) device, MOLLY See also (trademark).

[0067] In some embodiments, the compositions described herein are administered therapeutically to a subject by topical administration. As used herein, "local administration" or "local delivery" refers to does not rely on the transport of the composition or agent through the vascular system to the intended target tissue or site of For example, the composition may be delivered by injection or implantation of the composition or agent. or can be delivered by injection or implantation of a device containing the composition or agent. In certain embodiments, after local administration in the vicinity of the target tissue or site, the composition or the drug, or one or more components thereof, is administered to an intended target tissue other than the site of administration. It can spread to any tissue or area.

[0068] In some embodiments, the composition can be stored at a temperature below 0° C. (e.g., −20° C. or −80° C.). In some embodiments, the composition can be formulated for storage at 4°C (at room temperature). In some embodiments, the composition can be stored for up to 2 years. (e.g., 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months) , 10 months, 11 months, 1 year, 1.5 years, or 2 years) and stored at 2-8°C (e.g., 4°C). Thus, in some embodiments, the compositions described herein may be formulated to The composition is stable when stored at 2-8°C (eg, 4°C) for at least one year.

[0069] In some embodiments, the pharmaceutical composition may be formulated as a solution. In this form, the composition may be prepared in a buffer solution at a concentration suitable for storage at, for example, 2 to 8°C (e.g., 4°C). It can be formulated as follows.

[0070] Compositions comprising one or more antibodies described herein may be formulated in immunoliposomal compositions. Such formulations can be prepared by methods known in the art. Liposomes with enhanced circulation time are described, for example, in U.S. Pat. No. 5,013,556. It is stated in the book.

[0071] In certain embodiments, the compositions are administered in implants and microencapsulated delivery systems. Formulate the compound with a carrier that protects it from rapid release, such as a controlled release formulation containing a Biodegradable, biocompatible polymers, such as ethylene vinyl acetate, poly(ethylene vinyl acetate), polyanhydrides, polyglycolic acid, collagen, polyorthoesters, and polylactic acid. Many methods for preparing such formulations are known in the art. For example, JR Robinson (1978) "Sustained and Controlled Release Drug Delivery" ry Systems,” Marcel Dekker, Inc., New York.

[0072] In some embodiments, administration of the antibodies described herein involves administering nucleic acids encoding the antibodies to This is achieved by administering to the animal the nucleic acid encoding the therapeutic antibody described herein. to deliver nucleic acids that can be used to express and produce antibodies within cells It can be incorporated into genetic constructs used as part of gene therapy protocols. Expression constructs for such components can be used in any therapeutically effective carrier, e.g., to express the component genes in It can be administered in any formulation or composition that can be effectively delivered to cells in vivo. Techniques include recombinant retroviruses, adenoviruses, adeno-associated viruses, and viral vectors containing the herpes simplex virus-1 (HSV-1); This involves inserting a gene of interest into a recombinant bacterial or eukaryotic plasmid. Plasmid DNA can be used to directly transfect cells. Cationic liposomes (lipofectin) or derivatized polylysine conjugates lysate, gramicidin S, artificial viral envelopes, or other such intracellular carriers, and delivery of gene constructs by direct injection or with the aid of CaPO4 precipitation, etc. (See, for example, WO 04 / 060407). Examples of locoviruses include pLJ, pZIP, pWE, and pEM, which are known to those skilled in the art. (e.g., Eglitis et al. (1985) Science 230:1395-1398; Danos and Mulligan (1988) Proc Natl Acad Sci USA 85:6460-6464; Wilson et al. (1988) Proc Natl Acad Sci USA 85:3014-3018; Armentano et al. (1990) Proc Natl Acad Sci USA 87:6141-614 5; Huber et al. (1991) Proc Natl Acad Sci USA 88:8039-8043; Ferry et al. (1991) Proc Natl Acad Sci USA 88:8377-8381; Chowdhury et al. (1991) Science 254:1802-18 05; van Beusechem et al. (1992) Proc Natl Acad Sci USA 89:7640-7644; Kay et al. (1992) Human Gene Therapy 3:641-647; Dai et al. (1992) Proc Natl Acad Sci USA 89 :10892-10895; Hwu et al. (1993) J Immunol 150:4104-4115; US Patent No. 4,868, 116 and 4,980,286; and the International Publication of PCT Publications International Publication No. 89 / 07136, International Publication No. 89 / 02468, International Publication No. 89 / 05345 and International Publication No. 92 / 07573 Another viral gene delivery system utilizes adenovirus-derived vectors. (e.g., Berkner et al. (1988) BioTechniques 6:616; Rosenfeld et al. (1991) Science 252:431-434; and Rosenfeld et al. (1992) Cell 68:143-155). Adenovirus type 5 strain dl324 or other adenovirus strains (e.g., Ad2, Ad Suitable adenoviral vectors derived from Ad3, Ad7, etc. are known to those skilled in the art. Yet another viral vector system useful for delivering genes is the adeno-associated virus (AAV). AAV). For example, Flotte et al. (1992) Am J Respir Cell Mol Biol 7:349-356 ; Samulski et al. (1989) J Virol 63:3822-3828; and McLaughlin et al. (1989) J See Virol 62:1963-1973.

[0073] In some embodiments, the compositions provided herein are in unit dosage form, Such unit dosage forms may be suitable for self-administration. May be provided in a vial, cartridge, pre-filled syringe or disposable pen Dosing devices such as those described in U.S. Pat. No. 6,302,855 The sensor can also be used, for example, with the injection systems described herein.

[0074] Suitable compositions of the compositions described herein that can treat or prevent disorders in a subject The dosage will depend, for example, on the age, sex, and weight of the subject to be treated, as well as the particular The amount of inhibitor compound can depend on a variety of factors, including, for example, the amount of the antibody described herein. Different doses of a composition containing the antibody may reduce fibrosis compared to doses of different formulations of the antibody. Other factors that may affect the dose administered to a subject may be required to treat a subject with Factors include, for example, the type or severity of the disorder. Other factors include, for example, the Other medical disorders affecting the subject concurrently or in the past, the subject's general health, the subject's inheritance, Genetic predisposition, diet, time of administration, excretion rate, drug combinations, and any other factors administered to the subject. Additionally, specific dosages and concentrations for any particular subject may be included. It is also understood that the dosage and treatment regimen may be adjusted based on the judgment of the treating physician. sea ​​bream.

[0075] The compositions described herein may be administered as a fixed dose or in milligrams per kilogram ( In some embodiments, the dose may also be administered in a 1 mg / kg (100 mg / kg) dose. Reduce or prevent the production of antibodies or other host immune responses against one or more antigen-binding molecules. Exemplary Doses of Antibodies, Such as the Compositions Described Herein For example, the range is 0.0001 to 100 mg / kg, 0.01 to 5 mg / kg, etc. mg / kg, 1~1000mg / kg, 1~100mg / kg, 0.5~50mg / kg , 0.1-100mg / kg, 0.5-25mg / kg, 1-20mg / kg, and 1 For example, doses may range from 0.1 mg / kg to 10 mg / kg of body weight. .3mg / kg, 0.5mg / kg, 1.0mg / kg, 2.0mg / kg, 3.0mg / kg, 4.0mg / kg, 5.0mg / kg, 10mg / kg or 20mg / kg body weight, or in the range of 1 to 20 mg / kg body weight. An exemplary treatment regimen is 1 week Once a year, once every two weeks, once every three weeks, once every four weeks, once a month, once every three months, Or, administer once every 3 to 6 months, or initially administer at short intervals (e.g., once a week). Once a month to once every 3 weeks), followed by extended intervals (e.g., once a month to once every 3-6 weeks) The drug is administered once a month.

[0076] The pharmaceutical solution may comprise a therapeutically effective amount of the compositions described herein. Such an effective amount may be: In part, the effect of the administered composition, or, if multiple agents are used, the composition and Easily determined by one of ordinary skill in the art based on the combined effect with one or more additional active agents The therapeutically effective amount of the compositions described herein may also be determined based on the individual's disease state, age, sex, and body weight, as well as a composition (and one) that elicits a desired response in the individual. or a plurality of additional active agents), e.g., the ability of the active agent to modify at least one state parameter. The effectiveness of the treatment may vary depending on factors such as the improvement of at least one symptom of a fibrotic disorder. For example, a therapeutically effective amount of the compositions described herein may be used to treat a particular disorder, and / or to treat a condition in the art. inhibiting any one of the symptoms of the specific disorders known in or described herein (including severe A therapeutically effective amount can also reduce the severity or eliminate the occurrence of and / or prevent the disease. , an amount in which any toxic or detrimental effects of the composition outweigh the therapeutically beneficial effects.

[0077] Suitable human doses of any of the compositions described herein may be determined, for example, in a Phase I dose escalation study. Further evaluation can be performed in, for example, van Gurp et al. (2008) Am J Transplantation 8(8):1711-1718; Hanouska et al. (2007) Clin Cancer Res 13(2, part 1):523-531; and Hetherington et al. (2006) Antimicrobial Agents and Chemotherapy 50(10): 349 See 9-3500.

[0078] The toxicity and therapeutic efficacy of the compositions can be assessed in cell cultures or experimental animals (e.g., as described herein). fibrotic disorders) by known pharmaceutical procedures in animal models of These procedures can be used to 50 (a dose lethal to 50% of the population) and YobiED 50 The dose that is therapeutically effective in 50% of the population can be determined. The dose ratio for therapeutic effect is the therapeutic index, and the LD 50 / ED 50 It can be expressed as a ratio of Compositions described herein that exhibit a therapeutic index are preferred. Although it is possible to design a delivery system that targets such compounds to the site of the affected tissue, Care should be taken to minimize potential damage to normal cells, thereby reducing side effects. It is.

[0079] Data obtained from cell culture assays and animal studies are needed to support the development of a novel drug for use in humans. Those skilled in the art will recognize that a range of doses can be used in formulating the compositions described herein. Suitable doses of the compositions described above generally provide an ED with little or no toxicity. 50 A set containing The dosage will vary depending on the dosage form used and the route of administration utilized. For the compositions described herein, a therapeutically effective amount is initially Doses can be estimated from cell culture assays. Doses can be determined in animal models and in cell culture. IC 50 (i.e., the concentration of antibody that achieves half-maximal inhibition of symptoms) Such information can be used to formulate the drug to achieve a concentration range suitable for human use. Plasma levels can be measured, for example, by rapid In some embodiments, the amount of ATP can be measured by liquid chromatography. For example, if local administration (e.g., to the eye or joint) is desired, cell culture or animal models may be used. Determination of the dose required to achieve a therapeutically effective concentration within the local area It is possible.

[0080] All publications, patent applications, patents, and other references mentioned herein are the property of their respective owners. Further, the materials, methods, and examples are merely illustrative. Unless otherwise defined, the terms used herein are illustrative and not limiting. All technical and scientific terms used are commonly understood by one of ordinary skill in the art to which this invention belongs. Methods and materials similar or equivalent to those described herein Although any of the above may be used in the practice or testing of the present invention, suitable methods and materials are described herein. is stated.

[0081] The present disclosure is further illustrated by the following examples, which are provided for illustrative purposes only. They should not be construed as limiting the scope or content of the disclosure in any way. It is not something that can be done. [Example]

[0082] method Generation of novel antibodies against α11β1

[0083] Rat immunization Wistar rats were immunized with recombinant human α11β1 protein. Immunoassays for target human and mouse proteins were performed using an ELISA assay. The immune response was tested. Animals that generated a good immune response were then subjected to cell fusion (by electrofusion). All fused cells were plated in a 96-well plate, and the supernatant was purified by soluble The antibodies were screened against human and mouse α11β1 by ELISA. The clones were counterscreened against human α1β1, α2β1, and α10β1. It specifically binds to human and mouse α11β1 and binds to α1β1, α2β1, and α1 Clones that did not bind to 0β1 were selected, subcloned, expanded, and cryopreserved. Purified antibodies were then produced from selected clones, and the heavy and light chain variable domains were isolated. Sequences were obtained from each purified antibody.

[0084] Rabbit immunization A cell-based monoclonal antibody platform was used to immunize rabbits. Rabbits were immunized with recombinant human α11β1 protein. Splenocytes from the immunized rabbits were selected. and then selected against human β1 to reduce the number of β1-specific B cell clones. The selected splenocytes were cultured for approximately one week, and the culture supernatant was screened for binding to human α11β1. The top results were sequenced and then probed with rabbit antibodies using a HEK cell line. The organism was recombinantly produced.

[0085] Mouse immunization Ten mice from five different strains were analyzed using human α11β1, mouse α11β1, and Immunized with an appropriate mixture of tolerance-breaking proteins. Plasma titers were measured against human and mouse α11 The β1 mixture was evaluated by ELISA. Popliteal lymph nodes, inguinal lymph nodes, and Iliac lymph nodes were harvested and ELISA-positive anti-human / mouse α11β1 hybridomas were used. The cells were grown and subjected to secondary screening against human and mouse α11β1, a control HIS protein. and counterscreened against human α11β1, α2β1, and α10β1. The supernatant IgG concentration was sufficient for functional screening. Selected hybridomas that met the criteria were cloned and analyzed to identify human and mouse α11β1 Clonal hybridomas against the The IgG was purified. The variable regions of the heavy and light chains of the selected hybridomas were then sequenced. It has been decided.

[0086] Phage library display Phage library display was used to generate a fully human anti-α11β1 antibody. The fully human anti-α11β1 antibody is a phage-displayed single-chain fragment variable (scFv) antigen-binding It was discovered using purified human and Three rounds of selection were performed against the mouse α11β1 antigen and against α10β1. Deselection was performed to enrich for α11 subunit-specific antibodies. The appropriate population is subcloned into a bacterial soluble expression vector, recombinant antibody expression is induced, and E. coli is cultured. Supernatants were screened for binding using an LISA assay. The antibody bearing the nucleotide sequence was sequenced and subsequently converted from scFv to IgG.

[0087] ELISA 0.25 μg / mL of target antigen (recombinant human or mouse α11β1) was incubated overnight at 4°C. The plates were washed (with 0.1% Tween -20 in PBS) and blocked (2% BSA and 0.05% PBS containing Tween-20), various antibody concentrations, and incubated at room temperature for 1 hour. The plates were then washed and incubated with biotinylated anti-rabbit / macro antibodies in buffer at a 1:1000 dilution. The plates were incubated with mouse / human IgG and incubated at room temperature for 1 hour. After washing, streptavidin-HRP was added at 1:200 in dilution buffer and incubated at room temperature for 1 The plate was incubated for 1 hour. Ultra TMB ELISA substrate solution was added. The plates were incubated on a plate shaker for 5 minutes. Stop solution was added to each well. The reaction was stopped by 500 rpm and the plates were read at 450 nm.

[0088] FACS Antibody binding to CHO-K1 200,000 cells (wild-type CHO-K1 cells or CHO expressing human α11) Incubate the cells (i.e., 100-K1 cells) with the desired concentration of each antibody in FACS buffer for 30 min at 4°C. The cells were then washed with FACS buffer and incubated at 1:10 for 30 min at 4°C. The cells were then washed and incubated with secondary antibody at 100% dilution in PB for 20 min at room temperature. The cells were fixed in 1% PFA in PBS. After washing again, the cells were analyzed by cytometry using FACS buffer. It was read in.

[0089] Antibody binding to HPF / MF Human lung fibroblasts (ScienCell) were cultured in complete fibroblast growth medium (ScienCell). The cells were cultured in a T-150 flask at 4°C (37°F) until they reached 80% confluence. The cells were washed and harvested using Accutase. Cells were cultured at 7,500 cells / cm in complete FGF. m 2 The cells were seeded in a T-150 flask with 1000 kJ / ml and cultured for 72 hours. Then, the cells were washed and incubated in serum-low The cells were starved for 24 hours in reduced medium. After starvation, cells were incubated with TGFβ-1 (R&D System The cells were treated with 1000μL of ... The cells were seeded onto a nickel-bottom plate. Heat-inactivated fetal bovine serum (Gibco) was used to culture the cells. The cells were then blocked at 4°C for 30 minutes. The cells were then incubated at 4°C for 30 minutes as indicated in each figure. The cells were incubated with a human anti-α11 antibody (Creative Biosciences, oLabs) was included as a positive control and the appropriate IgG isotype as a negative control. The cells were washed twice and incubated at 4°C for 30 min with the IgG class specific anti-α11 antibody being tested. After incubation with PE-conjugated secondary antibody, cells were washed twice and fixed with 1% PFA for 30 minutes. The cells were then bound to FACS Verse (Benton Dickson) for each antibody. Data were acquired by gating on single cells and calculating the geometric mean of the PE channel for each sample. The fluorescence intensity (gMFI) was determined.

[0090] Surface Plasmon Resonance (SPR) The affinity of the antibody to human α11β1 was determined by surface plasmon resonance (SPR) assay. Affinity was measured using a Biacore T200 instrument at pH 7.6 and 25°C. The anti-HIS antibody was attached to the SPR sensor surface using EDC / NHS covalent binding. HIS-tagged human α11β1 was captured on the sensor surface and subjected to single-cycle ELISA. A kinetic assay was used. Increasing concentrations of α11β1 were applied to the sensor-bound α11β1. The test antibodies were injected serially. Dissociation was monitored for 1000 seconds. Data were double-reference subtracted using duplicate blank injected sensor surfaces. The Langmuir model of 1 was fitted to the data to estimate the kinetic association and dissociation constants. The affinity of the interaction (equilibrium dissociation constant) was calculated by dividing the kinetic dissociation constant by the kinetic binding constant. During the injection cycle, α11β1 and the bound antibody were analyzed by 1:1 at pH 1.5. It was cleared by injection of 0 mM glycine.

[0091] Cell adhesion inhibition 0.6×10 6 Cells / mL were incubated with various concentrations of each antibody for 20 minutes at 37°C. The cells were coated overnight at room temperature with 100 ng / mL type I collagen or PBS. The E-Plate VIEW 96 PET plate was incubated in 3% BSA for 1 hour at room temperature. After washing the plate with PBS, the mixture of cells and antibodies was added to the wells. and the plate was placed in the xCelligence instrument. Cell attachment was monitored for 6 hours. The time point of maximum cell adhesion was used for comparison with the control.

[0092] Fibroblast-to-myofibroblast transition (FMT) Human lung fibroblasts (ScienCell) were cultured in complete fibroblast growth medium (ScienCell). The cells were cultured in a 500 ml PBS (1000 ml) until they reached 80% confluence. Cells were plated in tissue culture-treated 96-well plates and incubated for 12 h at 4°C for 1 h at 4°C. After 24 hours, cells were washed and resuspended in serum-reduced medium. After starvation, the cells were incubated in TG medium with or without anti-α11 antibody. Fβ-1 (R&D Systems). Polyclonal rabbit anti-human α Eleven antibodies were used as positive controls. Appropriate IgG isotype controls were also included. After 48 hours, cells were harvested, fixed, permeabilized, and incubated with AlexaFluor 488-conjugated antibodies. Cells were stained with α-smooth muscle actin (αSMA) (Invitrogen). Verse (Benton Dickson) and determine the expression level of αSMA Data were gated on single cells and the geometric mean of the FITC channel was calculated for each sample. The gMFI of each sample was analyzed by determining the mean mean fluorescence intensity (gMFI). Normalized to control and expressed as % inhibition.

[0093] Collagen gel contraction assay The 24-well plates were blocked with 2% BSA in PBS overnight at 37°C. Before use in the assay, the plates were washed three times with PBS. HO cell lines were harvested and cultured in ExpiCHO Expression Medium (Gibco™ Catalog No. A29 10002) to 1.25 x 10 6 The collagen gel solution was resuspended in 100 ml of CHO cells. 3 mg / mL stock collagen type I (Gibco™ Collagen) in medium containing Gen I Rat Protein, Tail (Cat. No. A1048301) The solution was prepared by diluting to 100 mg / mL. Sodium hydroxide was added to this solution to adjust the pH. After neutralization, 400 μL of collagen solution was added to each well of a 24-well plate. For wells containing antibodies in the antibody gel solution, 2.5 × 10 CHO cells were 6 Individual adjustment The antibody was prepared in ExpiCHO medium at a final concentration of 2x. After combining 1:1, stock collagen type 1 was added. The gel was incubated at 37°C for 60 minutes. Antibodies were added to the ExpiCHO medium and layered on top of the polymerized gel (400 μL Gels were incubated at 37°C for 6 days, after which gel contraction was quantified. Analyze the images of each well using geJ and calculate gel shrinkage as a percentage of the initial gel area. It was decided as follows.

[0094] Tumor xenograft model Fifty-six female CB-17 SCID mice were transfected with A549 cells (5 × 10 6 Cells / Ma The tumor volume was approximately 100 mm 3 When this is reached, each animal is divided into 8 groups. Mice were randomly divided into seven groups of 10 mice each. Mice were then treated with either an isotype control or The novel mAbs 79E3E3, 16E10 and 9G04 (2 mg / kg and 20 mg / kg) administered intraperitoneally every 3 days for a total of 7 doses, or docetaxel at 10 mg / kg The tumor volume and body weight were measured twice a week between the two measurements. At 2-3 day intervals, the following conditions were met: weight loss of 20% or more; tumors that interfere with normal physiological functions such as migration; ulcerative tumors; 2000mm3 of The clinical observations of any of the following are observed: tumor size exceeding 100% and weakness, paralysis, seizures, and bleeding. Recorded until discovered.

[0095] Thinly sliced ​​liver slices (PCLS) Thinly sliced ​​liver slices (PCLS) were prepared from the excised liver tissue and cut before the start of the experiment. A post-stress period was allowed to elapse for 24 hours. (Group 1), or 100 μg / mL of control antibody (Groups 2 and 3 - mouse IgG2a or rabbit IgG), or TGF-β1 (3 ng / mL) and PD PCLS were cultured with a combination of GF-ββ (50 ng / mL) (groups 2 to 10). As a control, 10 μM Alk5i (group 4) was administered in the presence or absence of 5 to 10 μM Alk5i. In the group, a novel inhibitor (16E1) was administered at two increasing doses (10 and 100 μg / mL). 0, 79E3E3, and 9G05). Each group contained n = 6 human PCLS prepared from a single human liver. Refresh the PCLS culture medium containing all stimuli and compounds and collect it at 24-h intervals. The cell culture supernatant (n = 2 / 3 paired wells) was collected every 24 hours and the soluble production was measured. All PCLS were harvested at 96 hours and then flash frozen for quantification.

[0096] Markers of liver injury (lactate dehydrogenase (LDH) and aspartate transactivation) Tissue culture levels of aminase (AST) and hepatocyte function / viability (albumin) Albumin secretion was quantified in all PCLS at all time points. Collagen in cell culture supernatants was quantified by ELISA as a marker of integrity and function. The levels of gen1a1, IL-6, hyaluronic acid, and Timp-1 were measured using R&D Duo Quantification was performed using a set ELISA kit.

[0097] Total RNA extraction from PCLS was performed for all samples. A Qiagen i kit was used for RNA extraction. RNA was reverse transcribed into cDNA and qP Used in CR, Col1a1, αSMA, TIMP-1, TGF-β1, IL-6 , and β-actin / GAPDH transcription levels were measured.

[0098] [Example 1] Generation of a novel monoclonal antibody against α11β1 and determination of binding affinity Antibody screening was carried out by immunizing rats and rabbits with recombinant human α11β1 and detecting human and mouse α11β1. The study was carried out by immunizing mice with both human α11 and human α11β1 antibodies. β1 monoclonal antibodies were generated (24 rabbit, 7 rat, 20 mouse). The sequences of the heavy and light chain variable regions of the mouse and rat antibodies were determined, and those of the rabbit antibodies were determined. The complete heavy and light chain sequences were determined for the antibody.

[0099] E showing exemplary binding of selected mouse monoclonal antibodies to recombinant human α11β1 The LISA results are shown in Figure 2A. Three of these mAbs were also shown in Figure 2B. It also bound to mouse α11β1.

[0100] Data will also be collected to determine whether the antibody of interest binds to the I domain of α11β1. We used the I domain of α11β1 produced in-house. E3, 8H8E9, and 6E5C11, respectively, as measured by ELISA. The mouse antibodies 10-F23, 10-L15, 7-O8, and 6- A12, 9-G05, and 9-E16, and rabbit antibodies 7-H12 and 2-D3 was also tested for binding to the α11β1 I domain produced in-house. and Figure 3B shows graphs depicting binding data from exemplary mAbs.

[0101] α11β1 belongs to the collagen receptor family and has relatively high homology with them. Therefore, the novel antibodies of the present invention are directed to α1β1, α2β1 and / or α10β1. Table 2 includes the results of cross-reactivity with other receptors. nothing.

[0102] [Table 2]

[0103] Integrins are large transmembrane receptors that exist in a variety of conformational configurations, and therefore novel Experiments were performed to confirm that the antibody also binds to cell-expressed α11β1. The CHO-K1 cell line, which endogenously expresses the β1 subunit of α11, stably expresses human α11. The gene was modified to express CHO-K1 hu α11.

[0104] Figures 4A and 4B show that α11β1 binds to human α11β1 (as tested by ELISA) and CH Selected rat mAbs also showed the ability to bind to α11β1 expressed on the surface of O-K1 cells. Figures 11A and 11B show the surface expression of CHO cells and mouse mAb. Selected rabbit, rat, and mouse mAbs that showed binding ability to α11β1 b, and human mAb. Furthermore, Figure 14 shows the α11β expressed on the surface of CHO cells. 1. However, as shown in Table 1, ELISA showed that it bound to α11β1, but not to cell-expressed α11 Some mAbs did not bind to β1.

[0105] Combined EC 50 The fluorescence cell sorting was performed on CHO-K1 hu α11β1 cells. The results are shown in Table 3. The six m Four of the Abs were EC 50 Although the results were in the low nanomolar range (8-P20, 8-G 15, 8-J17, and 8-l14), whereas the remaining two mAbs were less potent (9 -G05 and 9-E16; both I-domain binders).

[0106] [Table 3]

[0107] As shown in Figure 9, antibodies 16E10, 79E3E3, 9G05, and 1994_01 C07 was tested for its affinity to human α11β1 by surface plasmon resonance (SPR). The KDs were 48 pM, 10 pM, 2.85 nM, and 0.77 nM, respectively. Interestingly, 16E10 and 1994_01_C07 have the I-dot structure of α11β1. Neither the main nor the headpiece domain bound to the ligand. It does not bind to the binding domain but acts allosterically by inhibiting α11β1 function. These results suggest that 9G05 and 79E3E3 may function as inhibitors of the I domain. can bind to the ligand-binding domain, thereby directly inhibiting the ligand-binding site The α11β1 headpiece and α11β1 I-domain as measured by SPR The binding affinities of the antibodies to the IgG4-dependent ATPases are shown in Figures 10A and 10B, respectively.

[0108] We also tested binding to physiologically relevant primary human cell types: human lung fibroblasts (HLF) Fibroblast-to-myofibroblast transition (FMT) using PF) and TGFβ treatment HPFs were induced to give rise to myofibroblasts (MFs). HPFs do not express α11β1, but Furthermore, HPFs express other collagen-binding receptors. HPF expresses the receptors α1β1 and α2β1, which means that HPF is a target for antibody cross-reactivity. This means that the reactivity of selected mAbs can be tested by HPF and Binding to MF was evaluated, and as shown in Figures 5, 12, and 13, 9- With the exception of E16, the antibodies tested did not bind to HPF but bound strongly to MF, which revealed some HPF binding, indicating off-target binding.

[0109] [Example 2] Biological activity of a novel monoclonal antibody against α11β1 Myofibroblasts secrete fibrous matrix, which prevents MF accumulation. The ablation and / or reduction of fibrosis is an important step in treating and / or preventing fibrosis. This was achieved by inhibiting the transition from fibroblasts to myofibroblasts using an anti-α11β1 antibody. This can be achieved by inhibiting ligand binding. Blocking the binding of α11β1 to type I collagen is a desirable feature of anti-α11β1 antibodies. Unlike many other receptors, Integrins mediate both "outside-in" (canonical ligand-mediated) and "outside-in" signaling. Therefore, inside-out signaling can be performed. affecting the structure of α11β1 in a manner that inhibits signal transduction and FMT, The antibody binds to α11β1 in a manner that does not affect its ability to bind to type I collagen. It is possible that α11β1 binds to α11β1. Therefore, blocking ligand binding Both blocking and non-blocking mAbs were included in these studies.

[0110] The CHO-K1 hu α11 cell line was used to investigate the α11β1-mediated response to type I collagen. The ability of mAbs to block binding was assessed. As shown in Figure 6A, the three mAbs tested Two of the rat mAbs inhibited CHO-K binding to type I collagen-coated plates. 1 hu α11 cell adhesion was significantly inhibited. Under the "no treatment" condition, the cells were transferred to type I colony. In the "no coating" condition, the cells were plated on collagen but no antibody was added. Cells were seeded onto BSA-coated wells without collagen type I. Statistics were performed using a single-agent method. Analysis of variance was performed using a multi-way analysis followed by Dunnett's multiple comparison test. The binder 79E3E3 blocks cell adhesion with an IC50 of 9.4 nM However, 40G10H11 strongly inhibited cell adhesion, whereas other collagens Although it cross-reacts with receptors (α1β1, α2β1, α10β1), it is an I-domain binder. It has not been confirmed that 24E4G6 binds to the I domain and not to collagen. When rabbit mAbs were tested, 8 out of 9 mAbs did not inhibit cell adhesion to the antibody. Three mAbs strongly and significantly inhibited cell adhesion, and none of them were I domain binders. Therefore, it was confirmed that these mAbs bind to the α11β1 domain. It is possible that integrins bind to the ATP-dependent ATPases, keeping them in a low- or intermediate-affinity state. Figure 1 shows the activity of selected mouse mAbs. Three of the six mAbs significantly blocked cell adhesion. Two of these mAbs were I-domain binders (9-G05 and 9- However, 8-G15 is a strong blocker of cell adhesion. However, binding to the I domain was not confirmed. 8-I14 did not block cell adhesion to type I collagen. The mAbs were analyzed for their ability to inhibit the binding of human CHO-α11 cells to type I collagen. As shown in Figures 15A and 15B, all human Abs were significantly higher than the control. In comparison, 1994-01-C07 inhibited cell adhesion, with an IC50 of 3.3 nM. As shown in these data (and outlined in Table 1), some anti-α1 1β1mAb binds strongly to human α11β1 as tested by ELISA and FACS Although it was confirmed that these antibodies block the ligand interaction, Furthermore, the I domain (the ligand-binding domain of α11β1) Binding to α11β1 is required to block the interaction between α11β1 and type I collagen. It was confirmed that this was not the case.

[0111] In addition to the above-mentioned binding ability, anti-α11β1 antibody inhibits the fibroblast-to-myofibroblast transition ( It is important to inhibit FMT. Myofibroblasts produce collagen extracellular matrix. Heterogeneous cells exist in various states of activation, with the primary function being the production and contraction of extracellular matrix (ECM). FMT is now recognized as a cell population that responds to changes in the mechanical environment of the repairing tissue. This is a multi-step event regulated by TGFβ, a potent factor enhancing this process. Alpha smooth muscle actin (αSMA) is one of the factors that contribute to the differentiation of fibroblasts into myofibroblasts. It is one of the major markers overexpressed during migration. The presence of αSMA is a key marker for intracellular fibroblast growth. Through a feedback loop, it promotes fibroblast contraction and induces myofibroblast activation. αSMA is a major molecular marker of myofibroblasts, and therefore, TGFβ-induced FM We tested the ability of a novel anti-α11β1 mAb to inhibit αSMA expression in T.

[0112] As shown in Figure 7A, two rat mAbs (40G10H11 and 24E4G6) , significantly inhibited αSMA expression compared to the control, but neither antibody was an I-domain binder. Statistics were determined by one-way analysis of variance followed by Dunnett's Furthermore, only 40G10H11 was found to be associated with type I collagen. Interestingly, 79E3E3 mAb inhibited cell adhesion of the I domain binder. It strongly inhibited cell adhesion to collagen, but suppressed αSMA expression (myofibroblast formation). As shown in Figure 7B, the two The Sagi mAb significantly inhibited αSMA expression compared to the control, but both antibodies inhibited the I domain. 16E10 was not confirmed to be an inbinder. It was confirmed that αSMA upregulation was inhibited (inhibition rate of 16G It was confirmed that 7 inhibits FMT but has no effect on cell adhesion to collagen. As shown in Figure 7C, five of the six murine mAbs tested significantly reduced α- Three FMT inhibitors (9-G05, 8-G15, 9-E) significantly inhibited the expression of SMA. 16) were also found to reduce cell adhesion to collagen, and two of them ( The mouse antibodies 8-J17 and 8- Only I14 inhibited FMT and had an effect on ligand binding.

[0113] [Example 3] Ability of selected antibodies to inhibit cell-mediated contraction of collagen gels Cell-mediated contraction of CD collagen I gels has previously been shown to be α11β1-mediated. This is a process that has been previously described, and more recent studies have shown that α11β1-mediated downstream signaling It was revealed that the ATP-dependent ATPase activity is essential for gel contraction to occur. The ability of the α-glucan-1-phosphate dehydrogenase (ADP) to inhibit α-glucan-mediated 3D gel contraction was tested because this ability is exemplary This is because it is directly related to the functionality of the antibody.

[0114] As shown in Figure 8, rat 79E3E3, mouse 9E16, 9G05, and 8I14 Rabbit 16E10, and human 1994_01_C07, 2004_04_B03, 20 All antibodies, 04_04_C12, and 1994_01_D12, were CHO-hu α Notably, CHO-hu α11 cells inhibited α11-mediated collagen gel contraction. As shown in the UT (untreated) condition, the cells were able to form collagen without the addition of TGFβ. Without treatment, cells were embedded in the collagen gel. No antibody was added. Statistics were calculated by one-way ANOVA followed by Dunnett's Multiple comparison tests were used. Each treatment condition was compared with the no treatment condition. Asterisks indicate statistical significance. The figures indicate statistical significance, and "ns" indicates that the difference was not statistically significant.

[0115] [Example 4] Assessment of the effects of selected antibodies on tumor xenograft growth Previous studies have shown that A549 cell xenotransplantation in α11 knockout SCID mice It has been shown that the growth of explants is significantly hindered compared to wild-type mice. In the present example, we investigated whether mAb-mediated inhibition of α11β1 function suppresses xenograft growth. As shown in Figure 16 and Table 4, a study was conducted to determine whether mouse CAFs expressed α11 Blocking β1 prevented the growth of xenografts in SCID mice. Specifically, 79E3E3, an effectorless mAb that cross-reacts with mouse α11β1, , which significantly suppressed tumor growth compared to the isotype control, but did not bind to murine α11β1. The non-targeting mAb 16E10 did not significantly inhibit tumor growth. Inhibition of β1 did not affect tumor growth, as 16E10 did not show any effect. There wasn't.

[0116] [Table 4]

[0117] [Example 5] Effect of anti-α11β1 antibody on human thin-sectioned liver slices (PCLS) Thinly sectioned liver slices (PCLS) from human liver tissue are physiological and structural representatives of tissue architecture. To evaluate the efficacy and clinical status of therapeutic targets by testing them in human PCLS. The relevance of the drug can be assessed in in vivo rodent models and in vitro. The limitations of 2D cell culture methods can be overcome. Tissue bioreactor technology is Viability and functionality of PCLS from human liver tissue were assessed for at least 6 days in vitro. Maintain.

[0118] As shown in Figures 17A-17C, all anti-α11-β1 antibodies tested inhibited soluble fibrinogen. Pro-fibrogenic markers (COL1A1, hyaluronan, TIMP1) were increased in a dose-dependent manner and The effect was partially inhibited at the highest dose tested. No toxicity was observed after treatment with either antibody. There were no elevations in ALT, AST, or albumin (data not shown). (Not at all).

[0119] Exemplary Sequences DNA sequence of anti-α11β1 monoclonal antibody Rat mAb sequence signal sequence-FR1-CDR1-FR2-CDR2-FR3-CDR3-FR4 79E3E3 heavy chain variable region signal sequence-FR1-CDR1-FR2-CDR2-FR3-CDR3-FR4

[0120] [ka]

[0121] 79E3E3 light chain variable region

[0122] [ka]

[0123] 24E4G6 heavy chain variable region

[0124] [ka]

[0125] 24E4G6 light chain variable region

[0126] [ka]

[0127] 8H8E9 heavy chain variable region

[0128] [ka]

[0129] 8H8E9 light chain variable region

[0130] [ka]

[0131] 6E5C11 heavy chain variable region

[0132] [ka]

[0133] 6E5C11 light chain variable region

[0134] [ka]

[0135] 7D8B10 heavy chain variable region

[0136] [ka]

[0137] 7D8B10 light chain variable region

[0138] [ka]

[0139] 18E10F10 heavy chain variable region

[0140] [ka]

[0141] 18E10F10 light chain variable region

[0142] [ka]

[0143] 40G10H11 heavy chain variable region

[0144] [ka]

[0145] 40G10H11 light chain variable region

[0146] [ka]

[0147] Mouse mAb sequence 9-G05 Heavy chain variable region

[0148] [ka]

[0149] 9-G05 Light chain variable region

[0150] [ka]

[0151] 8-P20 heavy chain variable region

[0152] [ka]

[0153] 8-P20 light chain variable region

[0154] [ka]

[0155] 8-G15 heavy chain variable region

[0156] [ka]

[0157] 8-G15 light chain variable region

[0158] [ka]

[0159] 8-I14 heavy chain variable region

[0160] [ka]

[0161] 8-I14 light chain variable region

[0162] [ka]

[0163] 9-E16 heavy chain variable region

[0164] [ka]

[0165] 9-E16 light chain variable region

[0166] [ka]

[0167] 8-J17 heavy chain variable region

[0168] [ka]

[0169] 8-J17 light chain variable region

[0170] [ka]

[0171] 6-O12 heavy chain variable region

[0172] [ka]

[0173] 6-O12 light chain variable region

[0174] [ka]

[0175] 10-L15 heavy chain variable region

[0176] [ka]

[0177] 10-L15 light chain variable region

[0178] [ka]

[0179] 7-H14 heavy chain variable region

[0180] [ka]

[0181] 7-H14 light chain variable region

[0182] [ka]

[0183] 6-B21 heavy chain variable region

[0184] [ka]

[0185] 6-B21 light chain variable region

[0186] [ka]

[0187] 10-F23 heavy chain variable region

[0188] [ka]

[0189] 10-F23 light chain variable region

[0190] [ka]

[0191] 6-A12 heavy chain variable region

[0192] [ka]

[0193] 6-A12 light chain variable region

[0194] [ka]

[0195] 6-M8 heavy chain variable region

[0196] [ka]

[0197] 6-M8 light chain variable region

[0198] [ka]

[0199] 2-A3 heavy chain variable region

[0200] [ka]

[0201] 2-A3 light chain variable region

[0202] [ka]

[0203] 6-O17 heavy chain variable region

[0204] [ka]

[0205] 6-O17 light chain variable region

[0206] [ka]

[0207] 3-G5 heavy chain variable region

[0208] [ka]

[0209] 3-G5 light chain variable region

[0210] [ka]

[0211] 6-A15 heavy chain variable region

[0212] [ka]

[0213] 6-A15 light chain variable region

[0214] [ka]

[0215] 10-K10 heavy chain variable region

[0216] [ka]

[0217] 10-K10 light chain variable region

[0218] [ka]

[0219] 6-P20 heavy chain variable region

[0220] [ka]

[0221] 6-P20 light chain variable region

[0222] [ka]

[0223] 7-O8 heavy chain variable region

[0224] [ka]

[0225] 7-O8 light chain variable region

[0226] [ka]

[0227] Rabbit mAb sequence A11B1_16G7 heavy chain

[0228] [ka]

[0229] A11B1_16G7 light chain

[0230] [ka]

[0231] A11B1_16E10 heavy chain

[0232] [ka]

[0233] A11B1_16E10 light chain

[0234] [ka]

[0235] A11B1_15G10 heavy chain

[0236] [ka]

[0237] A11B1_15G10 light chain

[0238] [ka]

[0239] A11B1_14H1 heavy chain

[0240] [ka]

[0241] A11B1_14H1 light chain

[0242] [ka]

[0243] A11B1_13G4 heavy chain

[0244] [ka]

[0245] A11B1_13G4 light chain

[0246] [ka]

[0247] A11B1_13C3 heavy chain

[0248] [ka]

[0249] A11B1_13C3 light chain

[0250] [ka]

[0251] A11B1_12F2 heavy chain

[0252] [ka]

[0253] A11B1_12F2 light chain

[0254] [ka]

[0255] A11B1_11D10 heavy chain

[0256] [ka]

[0257] A11B1_11D10 light chain

[0258] [ka]

[0259] A11B1_10F9 heavy chain

[0260] [ka]

[0261] A11B1_10F9 light chain

[0262] [ka]

[0263] A11B1_7H12 heavy chain

[0264] [ka]

[0265] A11B1_7H12 light chain

[0266] [ka]

[0267] A11B1_7G12 heavy chain

[0268] [ka]

[0269] A11B1_7G12 light chain

[0270] [ka]

[0271] A11B1_6G4 heavy chain

[0272] [ka]

[0273] A11B1_6G4 light chain

[0274] [ka]

[0275] A11B1_6F9 heavy chain

[0276] [ka]

[0277] A11B1_6F9 light chain

[0278] [ka]

[0279] A11B1_6C7 heavy chain

[0280] [ka]

[0281] A11B1_6C7 light chain

[0282] [ka]

[0283] A11B1_6B6 heavy chain

[0284] [ka]

[0285] A11B1_6B6 light chain

[0286] [ka]

[0287] A11B1_5F7 heavy chain

[0288] [ka]

[0289] A11B1_5F7 light chain

[0290] [ka]

[0291] A11B1_5D7 heavy chain

[0292] [ka]

[0293] A11B1_5D7 light chain

[0294] [ka]

[0295] A11B1_5A7 heavy chain

[0296] [ka]

[0297] A11B1_5A7 light chain

[0298] [ka]

[0299] A11B1_4E1 heavy chain

[0300] [ka]

[0301] A11B1_4E1 light chain

[0302] [ka]

[0303] A11B1_3H9 heavy chain

[0304] [ka]

[0305] A11B1_3H9 light chain

[0306] [ka]

[0307] A11B1_3G2 heavy chain

[0308] [ka]

[0309] A11B1_3G2 light chain

[0310] [ka]

[0311] A11B1_3B1 heavy chain

[0312] [ka]

[0313] A11B1_3B1 light chain

[0314] [ka]

[0315] A11B1_2D3 heavy chain

[0316] [ka]

[0317] A11B1_2D3 light chain

[0318] [ka]

[0319] A11B1_2A7 heavy chain

[0320] [ka]

[0321] A11B1_2A7 light chain

[0322] [ka]

[0323] Protein sequence of anti-α11β1 monoclonal antibody Rat mAb sequence signal peptide-FR1-CDR1-FR2-CDR2-FR3-CDR3-FR4 79E3E3 heavy chain variable region

[0324] [ka]

[0325] 79E3E3 light chain variable region

[0326] [ka]

[0327] 24E4G6 heavy chain variable region

[0328] [ka]

[0329] 24E4G6 light chain variable region

[0330] [ka]

[0331] 8H8E9 heavy chain variable region

[0332] [ka]

[0333] 8H8E9 light chain variable region

[0334] [ka]

[0335] 6E5C11 heavy chain variable region

[0336] [ka]

[0337] 6E5C11 light chain variable region

[0338] [ka]

[0339] 7D8B10 heavy chain variable region

[0340] [ka]

[0341] 7D8B10 light chain variable region

[0342] [ka]

[0343] 18E10F10 heavy chain variable region

[0344] [ka]

[0345] 18E10F10 light chain variable region

[0346] [ka]

[0347] 40G10H11 heavy chain variable region

[0348] [ka]

[0349] 40G10H11 light chain variable region

[0350] [ka]

[0351] Mouse mAb sequence FR1-CDR1-FR2-CDR2-FR3-CDR3-FR4 9-G05 Heavy chain variable region

[0352] [ka]

[0353] 9-G05 Light chain variable region

[0354] [ka]

[0355] 8-P20 heavy chain variable region

[0356] [ka]

[0357] 8-P20 light chain variable region

[0358] [ka]

[0359] 8-G15 heavy chain variable region

[0360] [ka]

[0361] 8-G15 light chain variable region

[0362] [ka]

[0363] 8-I14 heavy chain variable region

[0364] [ka]

[0365] 8-I14 light chain variable region

[0366] [ka]

[0367] 9-E16 heavy chain variable region

[0368] [ka]

[0369] 9-E16 light chain variable region

[0370] [ka]

[0371] 8-J17 heavy chain variable region

[0372] [ka]

[0373] 8-J17 light chain variable region

[0374] [ka]

[0375] 9-B11 heavy chain variable region

[0376] [ka]

[0377] 9-B11 light chain variable region

[0378] [ka]

[0379] For SEQ ID NOs: 117-144, CDR3 is shown in bold and underlined.

[0380] 6-O12 heavy chain variable region

[0381] [ka]

[0382] 6-O12 light chain variable region

[0383] [ka]

[0384] 10-L15 heavy chain variable region

[0385] [ka]

[0386] 10-L15 light chain variable region

[0387] [ka]

[0388] 7-H14 heavy chain variable region

[0389] [ka]

[0390] 7-H14 light chain variable region

[0391] [ka]

[0392] 6-B21 heavy chain variable region

[0393] [ka]

[0394] 6-B21 light chain variable region

[0395] [ka]

[0396] 10-F23 heavy chain variable region

[0397] [ka]

[0398] 10-F23 light chain variable region

[0399] [ka]

[0400] 6-A12 heavy chain variable region

[0401] [ka]

[0402] 6-A12 light chain variable region

[0403] [ka]

[0404] 6-M8 heavy chain variable region

[0405] [ka]

[0406] 6-M8 light chain variable region

[0407] [ka]

[0408] 2-A3 heavy chain variable region

[0409] [ka]

[0410] 2-A3 light chain variable region

[0411] [ka]

[0412] 6-O17 heavy chain variable region

[0413] [ka]

[0414] 6-O17 light chain variable region

[0415] [ka]

[0416] 3-G5 heavy chain variable region

[0417] [ka]

[0418] 3-G5 light chain variable region

[0419] [ka]

[0420] 6-A15 heavy chain variable region

[0421] [ka]

[0422] 6-A15 light chain variable region

[0423] [ka]

[0424] 10-K10 heavy chain variable region

[0425] [ka]

[0426] 10-K10 light chain variable region

[0427] [ka]

[0428] 6-P20 heavy chain variable region

[0429] [ka]

[0430] 6-P20 light chain variable region

[0431] [ka]

[0432] 7-O8 heavy chain variable region

[0433] [ka]

[0434] 7-O8 light chain variable region

[0435] [ka]

[0436] Rabbit mAb sequence A11B1_16G7 heavy chain

[0437] [ka]

[0438] A11B1_16G7 light chain

[0439] [ka]

[0440] A11B1_16E10 heavy chain

[0441] [ka]

[0442] A11B1_16E10 light chain

[0443] [ka]

[0444] A11B1_15G10 heavy chain

[0445] [ka]

[0446] A11B1_15G10 light chain

[0447] [ka]

[0448] A11B1_14H1 heavy chain

[0449] [ka]

[0450] A11B1_14H1 light chain

[0451] [ka]

[0452] A11B1_13G4 heavy chain

[0453] [ka]

[0454] A11B1_13G4 light chain

[0455] [ka]

[0456] A11B1_13C3 heavy chain

[0457] [ka]

[0458] A11B1_13C3 light chain

[0459] [ka]

[0460] A11B1_12F2 heavy chain

[0461] [ka]

[0462] A11B1_12F2 light chain

[0463] [ka]

[0464] A11B1_11D10 heavy chain

[0465] [ka]

[0466] A11B1_11D10 light chain

[0467] [ka]

[0468] A11B1_10F9 heavy chain

[0469] [ka]

[0470] A11B1_10F9 light chain

[0471] [ka]

[0472] A11B1_7H12 heavy chain

[0473] [ka]

[0474] A11B1_7H12 light chain

[0475] [ka]

[0476] A11B1_7G12 heavy chain

[0477] [ka]

[0478] A11B1_7G12 light chain

[0479] [ka]

[0480] A11B1_6G4 heavy chain

[0481] [ka]

[0482] A11B1_6G4 light chain

[0483] [ka]

[0484] A11B1_6F9 heavy chain

[0485] [ka]

[0486] A11B1_6F9 light chain

[0487] [ka]

[0488] A11B1_6C7 heavy chain

[0489] [ka]

[0490] A11B1_6C7 light chain

[0491] [ka]

[0492] A11B1_6B6 heavy chain

[0493] [ka]

[0494] A11B1_6B6 light chain

[0495] [ka]

[0496] A11B1_5F7 heavy chain

[0497] [ka]

[0498] A11B1_5F7 light chain

[0499] [ka]

[0500] A11B1_5D7 heavy chain

[0501] [ka]

[0502] A11B1_5D7 light chain

[0503] [ka]

[0504] A11B1_5A7 heavy chain

[0505] [ka]

[0506] A11B1_5A7 light chain

[0507] [ka]

[0508] A11B1_4E1 heavy chain

[0509] [ka]

[0510] A11B1_4E1 light chain

[0511] [ka]

[0512] A11B1_3H9 heavy chain

[0513] [ka]

[0514] A11B1_3H9 light chain

[0515] [ka]

[0516] A11B1_3G2 heavy chain

[0517] [ka]

[0518] A11B1_3G2 light chain

[0519] [ka]

[0520] A11B1_3B1 heavy chain

[0521] [ka]

[0522] A11B1_3B1 light chain

[0523] [ka]

[0524] A11B1_2D3 heavy chain

[0525] [ka]

[0526] A11B1_2D3 light chain

[0527] [ka]

[0528] A11B1_2A7 heavy chain

[0529] [ka]

[0530] A11B1_2A7 light chain

[0531] [ka]

[0532] Human mAb sequence Heavy and light chain variable region sequences 2004_04_B03

[0533] [Table 5]

[0534] 2004_05_A06

[0535]

Table 6

[0536] 2004_04_C12

[0537]

Table 7

[0538] 2002_02_B07

[0539]

Table 8

[0540] 2004_05_B04

[0541]

Table 9

[0542] 2003_03_E12

[0543]

Table 10

[0544] 1994_01_C07

[0545]

Table 11

[0546] 1995_01_G07

[0547]

Table 12

[0548] 1995_01_G05

[0549]

Table 13

[0550] 2004_03_G10

[0551]

Table 14

[0552] 2002_02_B05

[0553]

Table 15

[0554] 2003_03_F05

[0555] Table 16

[0556] 1994_01_A07

[0557] Table 17

[0558] 1994_01_A09

[0559] Table 18

[0560] 1994_01_D12

[0561] Table 19

[0562] 1995_01_F05

[0563] Table 20

[0564] 1995_01_F09

[0565] Table 21

[0566] 1996_01_H07

[0567] Table 22

[0568] 1997_02_B01

[0569] Table 23

[0570] 2002_02_E01

[0571] Table 24

[0572] 2002_02_G11

[0573] Table 25

[0574] 2003_03_A09

[0575] Table 26

[0576] 2004_04_D03

[0577] [Table 27]

[0578] 2004_04_F01

[0579] [Table 28]

[0580] 2005_05_E05

[0581] [Table 29]

[0582] 1994_01_D04

[0583] [Table 30]

[0584] 1997_02_B03

[0585] [Table 31]

[0586] Humanized mAb sequence Humanized 79E3E3 heavy chain variable region

[0587] [ka]

[0588] Humanized 79E3E3 light chain variable region

[0589] [ka]

[0590] Humanized 9-G05 heavy chain VH_1 Leader sequence-VH-hIgG1CH-stop codon*

[0591] [ka]

[0592] Humanized 9-G05 heavy chain VH_2 Leader sequence-VH-hIgG1CH-stop codon*

[0593] [ka]

[0594] Humanized 9-G05 heavy chain VH_3 Leader sequence-VH-hIgG1CH-stop codon*

[0595] [ka]

[0596] Humanized 9-G05 heavy chain VH_4 Leader sequence-VH-hIgG1CH-stop codon*

[0597] [ka]

[0598] Humanized 9-G05 light chain VL_1 Leader sequence-VL-hIgKCL-stop codon*

[0599] [ka]

[0600] Humanized 9-G05 light chain VL_2 Leader sequence-VL-hIgKCL-stop codon*

[0601] [ka]

[0602] 16E10 heavy chain variable region

[0603] [ka]

[0604] Humanized 16E10 heavy chain variable region VH_1

[0605] [ka]

[0606] Humanized 16E10 heavy chain variable region VH_2

[0607] [ka]

[0608] Humanized 16E10 heavy chain variable region VH_3

[0609] [ka]

[0610] Humanized 16E10 heavy chain variable region VH_4

[0611] [ka]

[0612] Humanized 16E10 heavy chain variable region VH_5

[0613] [ka]

[0614] Humanized 16E10 heavy chain variable region VH_6

[0615] [ka]

[0616] 16E10 light chain variable region

[0617] [ka]

[0618] Humanized 16E10 light chain variable region VL_1

[0619] [ka]

[0620] Humanized 16E10 light chain variable region VL_2

[0621] [ka]

[0622] Humanized 16E10 light chain variable region VL_3

[0623] [ka]

[0624] Humanized 16E10 light chain variable region VL_4

[0625] [ka]

[0626] Humanized 16E10 light chain variable region VL_5

[0627] [ka]

[0628] Humanized 16E10 light chain variable region VL_6

[0629] [ka]

[0630] equivalent Although the present invention has been described in detail, the foregoing description is intended to illustrate the scope of the present invention. and is not intended to limit the scope of the invention, which is set forth in the appended claims. It should be understood that the scope of the present invention is defined by the following: It is within the scope of the claims.

Claims

1. an anti-integrin antibody comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 103 to 435; An anti-alpha 11 beta 1 (α11β1) antibody or an antigen-binding fragment thereof.

2. SEQ ID NOs: 103 to 207, 209, 211, 213, 216, 218, 220, and 223 、２２５、２２８、２３３、２３４、２３６、２４０、２４１、２４５、２４７、２５３ 、２５５、２５７、２５９、２６１、２６５、２６７、２６９、２７１、２７５、２７７ 、２７９、２８１、２８３、２８７、２８９、２９１、２９３、２９６、３００、３０４ 、３０６、３０８、３１０、３１２、３１４、３１６、３１８、３２０、３２２、３２４ 、３２５、３２７、３２９、３３４、３３６、３３８、３４０、３４２、３４４、３４８ 、３５１、３５３、３５５、３５８、３６０、３６１、３６４、３６６、３６８、３６９ 、３７４、３７６、３７７、３７９、３８０、３８１、３８３、３８４、３８５、３８７ 、３８９、３９２、３９３、３９６、３９８、４００、４０２、４０５、４０８、４１１ or an anti-α11β1 antibody comprising a CDR sequence included in any one of 413 to 435. The body or an antigen-binding fragment thereof.

3. a CDR1 encompassed by any one of SEQ ID NOs: 103-206 or 413-435; An anti-α11β1 antibody or antigen-binding fragment thereof, comprising CDR2 and CDR3.

4. Selected from the group consisting of SEQ ID NOs: 103-114, 207-311, and 312-435 The anti-α11β1 antibody or antigen-binding fragment thereof according to claim 1, comprising an amino acid sequence as defined by Piece.

5. SEQ ID NOs: 103 to 114, 207, 209, 211, 213, 216, 218, 220 、２２３、２２５、２２８、２３３、２３４、２３６、２４０、２４１、２４５、２４７ 、２５３、２５５、２５７、２５９、２６１、２６５、２６７、２６９、２７１、２７５ 、２７７、２７９、２８１、２８３、２８７、２８９、２９１、２９３、２９６、３００ 、３０４、３０６、３０８、３１０、３１２、３１４、３１６、３１８、３２０、３２２ 、３２４、３２５、３２７、３２９、３３４、３３６、３３８、３４０、３４２、３４４ 、３４８、３５１、３５３、３５５、３５８、３６０、３６１、３６４、３６６、３６８ 、３６９、３７４、３７６、３７７、３７９、３８０、３８１、３８３、３８４、３８５ 、３８７、３８９、３９２、３９３、３９６、３９８、４００、４０２、４０５、４０８ 411, or 413-435.

3. An anti-α11β1 antibody or an antigen-binding fragment thereof according to 2.

6. one encompassed by any one of SEQ ID NOs: 103-114 or 413-434; or The anti-α11β1 antibody or antigen-binding fragment thereof according to claim 2, comprising multiple CDR sequences. 。

7. a CDR1 encompassed by any one of SEQ ID NOs: 103-114 or 413-434; The anti-α11β1 antibody or its antigen binding domain according to claim 3, comprising CDR2 and CDR3. Synthetic fragment.

8. The antibody according to any one of claims 1 to 7, which is a monoclonal antibody or an antigen-binding fragment thereof. The anti-α11β1 antibody or antigen-binding fragment thereof described above.

9. The antibody according to any one of claims 1 to 8, which is a humanized antibody or an antigen-binding fragment thereof. An α11β1 antibody or an antigen-binding fragment thereof.

10. The present invention relates to a method for reducing the interaction between α11β1 and collagen in human α11β1-expressing cells. The anti-α11β1 antibody or antigen-binding fragment thereof according to any one of claims 1 to 9.

11. An antibody or antigen-binding fragment thereof that competes with the antibody or antigen-binding fragment thereof according to any one of claims 1 to 10. An α11β1 antibody or an antigen-binding fragment thereof.

12. A nucleic acid encoding the antibody or antigen-binding fragment thereof according to any one of claims 1 to 11. A nucleic acid, including a sequence of nucleic acids.

13. Claim 11: The nucleic acid sequence comprises a sequence selected from the group consisting of SEQ ID NOs: 1 to 102 The nucleic acid according to

14. A vector comprising the nucleic acid of claim 12 or 13.

15. A host cell comprising the nucleic acid of claim 12 or 13 or the vector of claim 14. Cell.

16. 15. The host cell of claim 14 under conditions suitable for expression of the antibody or antigen-binding fragment thereof. A method for producing an antibody or an antigen-binding fragment thereof, comprising culturing the antibody or antigen-binding fragment thereof.

17. A method of treating a subject having or at risk of having a fibrotic disorder, comprising administering to a subject a therapeutically effective amount of 12. A method for treating a subject with an effective amount of the antibody or antigen-binding fragment thereof according to any one of claims 1 to 11. The method of claim 1, further comprising administering to

18. The fibrotic disorder may be idiopathic pulmonary fibrosis (IPF), chronic kidney disease, diabetic cardiomyopathy, primary cirrhosis, or Primary biliary cholangitis (PSC), primary biliary cirrhosis (PBC), nonalcoholic fatty liver disease ( NAFLD / NASH), Crohn's disease, ulcerative colitis, or systemic sclerosis, Item 18. The method according to item 17.

19. 1. A method of treating a subject having or at risk of cancer, comprising administering a therapeutically effective amount of 12. A method for treating a subject comprising administering to said subject an antibody or antigen-binding fragment thereof according to any one of claims 1 to 11. The method includes providing.

20. The cancer is head and neck squamous cell carcinoma, pancreatic ductal adenocarcinoma, non-small cell lung cancer, adrenocortical carcinoma, acute myeloid leukemia, Blood disease, bladder urothelial cancer, invasive breast cancer, cervical squamous cell carcinoma, bile duct cancer, colorectal adenocarcinoma, Large B-cell lymphoma, esophageal adenocarcinoma, glioblastoma multiforme, hepatocellular carcinoma, lung adenocarcinoma , lung squamous cell carcinoma, skin melanoma, mesothelioma, ovarian serous cystadenocarcinoma, pheochromocytoma and paraganglioma tumor, prostate adenocarcinoma, sarcoma, gastric adenocarcinoma, testicular germ cell tumor, thymoma, thyroid cancer, endometrial carcinoma of the uterine corpus carcinoma, uterine carcinosarcoma, uveal melanoma, renal clear cell carcinoma, renal carcinosarcoma, renal papillary cell carcinoma, or 20. The method of claim 19, wherein the hydroxyl group is a hydroxyl group or a combination thereof.

21. further comprising administering to said subject an effective amount of a chemotherapeutic or oncolytic therapeutic agent.

21. The method according to claim 19 or 20.

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