Integrin alpha 10 antibody

Abstract

The present invention relates to novel integrin alpha O antibodies and their use in medicine. 25 humanized antibodies (abs) derived from known mouse mAb365 (hybridoma deposited under accession number DSM ACC2583) are disclosed. Those that specifically bind to integrin alpha10 beta1 were selected. Five of the 25 humanized variants were selected as lead candidates (designated TAR-Ab8, TAR-Ab9, TAR-Abl3, TAR-Abl4, and TAR-Ab23) because all five antibodies showed improved thermal stability compared to a chimeric antibody designated AR-Ab0. Only TAR-Ab23 shows the additional functional property of having a higher binding affinity than mAb365.

Description

[Technical field]

[0001] The present invention relates to novel integrin alpha10 antibodies. [Background technology]

[0002] Integrins constitute a major family of cell adhesion receptors and are essential for a wide range of physiological processes, including the formation and maintenance of tissue architecture, cell migration, proliferation, and differentiation. Integrins also play an important role in tumor growth and metastasis. Integrins are assembled from 18 different alpha and 8 different beta subunits, which consist of one alpha and one beta subunit and form 24 functionally distinct integrin heterodimers in mammals. Integrins have a large extracellular region consisting of a ligand-binding globular head constructed from both alpha and beta subunits and two legs connected to a single transmembrane helix and a short cytoplasmic domain (Hohenster 2014). Some alpha subunits, including integrin alpha10, have an I-domain (collagen-binding region) inserted into the globular head.

[0003] Integrin alpha 10 beta 1 Integrin alpha10 (gene name ITGA10) belongs to the collagen-binding integrin subfamily consisting of alpha1, alpha2, alpha10, and alpha11 (Gullberg and Lundgren-Askerlund 2002). Sequence analysis shows that alpha10 has the highest identity (43%) with alpha11, 33% with alpha1, and 31% with alpha2. Integrin alpha10 beta1 is expressed primarily on chondrocytes in articular cartilage, cartilage supporting the vertebral column, trachea, and bronchi (Camper et al. 2001), as well as on specialized fibrous tissues such as the endosteum (cells lining the space between the bone marrow) and periosteum (cells lining the outside of the bone), the fascia of skeletal muscle and tendons, and on some cells within the ossification groove of Ranvier that may represent mesenchymal stem cells known to reside in these tissues (Camper et al. 2001; Gullberg and Lundgren-Akerlund 2002; Varas L et al. 2007; Lundgren-Akerlund and Aszodi 2014). Integrin alpha10 beta1 has also been found in isolated mesenchymal stem cells (Varas L et al. 2007; Uvebrant et al. 2019). The interaction between integrin alpha10beta1 and collagen, as well as other integrin-extracellular matrix protein interactions, mediates specific signaling between cells and the surrounding extracellular matrix (Frith et al. 2012, Campbell et al. 2011, Heino 2000, Bondreau and Jones 1999, Hering 1999). Knockout studies in mice have demonstrated that integrin alpha10beta1 is important for growth plate formation and long bone growth during skeletal development (Bengtsson T et al. 2005). More recently, analysis of an inbred dog line exhibiting a truncated mutation in ITGA10 demonstrated complete canine chondrodysplasia (Kyyostilai et al. 2013).

[0004] Integrin alpha 10 beta 1 antibodies for cancer treatment Interactions between integrin receptors on tumor cells and extracellular matrix molecules have been shown to play a key role in tumor cell functions such as cell migration, invasion, proliferation, and survival (Kechagia, JZ et al., 2019; Moreno-Layseca, P et al., 2019; Malric, L et al., 2017). Although information on the role of integrin α10β1 in cancer is still limited, recent studies have shown that integrin α10β1 is important for tumor growth and metastasis (Thoren et al. 2019; Masoumi et al., 2021.; Ellert-Miklaszewska et al., 2020). Furthermore, integrin α10β1 represents a promising target for directed antibody-based cancer therapy due to its high cell surface expression in certain cancers such as TNBC and glioblastoma, and very low expression in most normal tissues. Accordingly, function-blocking integrin alpha10 beta1 antibodies have been shown to reduce cancer cell migration, growth, and proliferation in preclinical cancer models of tumors of the CNS (WO2016 / 133449), breast cancer, particularly triple-negative breast cancer, prostate cancer, pancreatic cancer, and lung cancer (WO2020 / 212416). Validation of cancer targets has been performed with mouse antibodies targeting human integrin alpha10 beta1, but these are too immunogenic for clinical use due to the generation of human anti-mouse antibodies. However, immunogenicity can be reduced through humanization of mouse monoclonal antibodies. Furthermore, the human constant region allows Fc-mediated effector functions and extends serum half-life. However, the binding affinity of humanized antibodies is often reduced compared to the original mouse antibody (Hideaki Sanada et al., 2018; Scientific Report; Riechmann L, 1988; Nature; Jones PT et al. 1986; Nature; Foote J et al., 1992, J. Mol. Biol).

[0005] The goal of this study is to generate a humanized version of a function-blocking murine antibody, mAb365, for potential future use as a therapeutic agent for the treatment of cancer. Summary of the Invention

[0006] The inventors of the present invention have developed a novel humanized integrin alpha10 antibody that has unexpected and advantageous characteristics compared to murine mAb365. The antibody is a humanized monoclonal antibody that specifically binds to the extracellular I domain of the integrin alpha10 subunit of integrin alpha10 beta1, and has the ability to inhibit the adhesion, proliferation, and migration of cancer cells, as well as to reduce tumor growth and metastasis.

[0007] Thus, in one aspect, the present invention relates to an antibody or antigen-binding fragment thereof having binding specificity for integrin alpha10, said antibody or antigen-binding fragment comprising: A light chain variable region, a) a CDR-L1 comprising or consisting of the amino acid sequence of SEQ ID NO: 1; b) a CDR-L2 comprising or consisting of an amino acid sequence selected from the group consisting of SEQ ID NO: 2 and SEQ ID NO: 4; and c) a light chain variable region comprising a CDR-L3 comprising or consisting of the amino acid sequence of SEQ ID NO:5; and / or A heavy chain variable region, a) a CDR-H1 comprising or consisting of an amino acid sequence selected from the group consisting of SEQ ID NO: 7, SEQ ID NO: 6 and SEQ ID NO: 8; b) a CDR-H2 comprising or consisting of the amino acid sequence of SEQ ID NO: 9; and c) a heavy chain variable region comprising a CDR-H3 comprising or consisting of the amino acid sequence of SEQ ID NO:10.

[0008] In another aspect, the invention relates to a polynucleotide encoding an antibody or antigen-binding fragment thereof as defined herein, or its component polypeptide chains.

[0009] In yet another aspect, the present invention relates to a vector comprising a polynucleotide as defined herein.

[0010] In yet another aspect, the present invention relates to a recombinant host cell comprising a polynucleotide as defined herein or a vector as defined herein.

[0011] In yet another aspect, the present invention relates to a pharmaceutical composition comprising an antibody or antigen-binding fragment thereof as defined herein, a polynucleotide as defined herein, a vector as defined herein, and / or a recombinant host cell as defined herein, wherein the composition further comprises a pharma- ceutically acceptable buffer, diluent, carrier, or excipient.

[0012] In yet another aspect, the invention relates to a method for producing an antibody or antigen-binding fragment thereof as defined herein, the method comprising culturing a host cell as defined herein comprising a polynucleotide as defined herein or a vector as defined herein under conditions allowing expression of the encoded antibody or antigen-binding fragment thereof.

[0013] In yet another aspect, the present invention relates to an antibody or antigen-binding fragment thereof as defined herein, a polynucleotide as defined herein, a vector as defined herein, a recombinant host cell as defined herein, and / or a composition as defined herein for use in medicine.

[0014] In yet another aspect, the present invention relates to an antibody or antigen-binding fragment thereof as defined herein, a polynucleotide as defined herein, a vector as defined herein, a recombinant host cell as defined herein, and / or a composition as defined herein for use in the prevention, treatment, amelioration, detection and / or diagnosis of a disease or disorder susceptible to treatment with an inhibitor of integrin alpha10 beta1 and / or which disease or disorder is associated with cells expressing integrin alpha10 beta1.

[0015] In yet another aspect, the present invention relates to an antibody or antigen-binding fragment thereof as defined herein, a polynucleotide as defined herein, a vector as defined herein, a recombinant host cell as defined herein, and / or a composition as defined herein for use in inhibiting cell migration, cell proliferation, cell growth, cell survival, and / or cell adhesion, wherein the target cell may be a cancer cell and / or a cancer-associated cell expressing integrin alpha10 beta1.

[0016] In yet another aspect, the present invention relates to an in vitro method for the detection of cells expressing integrin alpha10 beta1 in a subject, the method comprising: (a) providing a sample of cells from the subject to be tested, such as a tissue biopsy or blood sample; (b) optionally extracting and / or purifying cells present in the sample; (c) contacting an antibody or antigen-binding fragment thereof as defined herein with cells present in the sample; (d) determining whether the antibody or antigen-binding fragment thereof binds to the cell; Binding of the antibody or antigen-binding fragment thereof to the cells indicates the presence of a disease or disorder associated with cells expressing integrin alpha10 beta1 in the tissue of the subject.

[0017] In yet another aspect, the present invention relates to an in vitro method for identifying a patient having a disease or disorder associated with cells expressing integrin alpha10 beta1 that would benefit from treatment with an antibody or antigen-binding fragment thereof as defined herein, the method comprising: a) providing a sample, such as a tissue biopsy or blood sample, from a patient to be tested; b) optionally extracting and / or purifying cells present in the sample; c) contacting a cell with an antibody or antigen-binding fragment thereof as defined herein; d) determining whether the antibody or antigen-binding fragment thereof binds to the integrin alpha 10 domain or a fragment thereof; Binding of the antibody or antigen-binding fragment thereof to the integrin alphal0 domain or a fragment thereof is indicative of a patient who will benefit from treatment with an antibody or antigen-binding fragment thereof as defined herein.

[0018] In yet another aspect, the present invention provides a method for treating a patient having a disease or disorder associated with cells expressing integrin alpha10 beta1, the method comprising: a) selecting a patient identified as having a disease or disorder associated with cells expressing integrin alpha10 beta1 as defined herein; b) administering to the patient a therapeutic agent effective to treat the disease or disorder.

[0019] In yet another aspect, the present invention relates to a method for the detection of cells expressing integrin alpha10 beta1, the method comprising: a) contacting an antibody or antigen-binding fragment thereof as defined herein with a cell to be analyzed for expression of integrin alpha10 beta1; b) determining whether the antibody or antigen-binding fragment thereof binds to the cell; Binding of the antibody or antigen-binding fragment thereof to the cells indicates the presence of a disease or disorder associated with cells expressing integrin alpha10 beta1 in the tissue of the subject.

[0020] In yet another aspect, the present invention provides a method for in vivo imaging of integrin alpha10 beta1 expression in a mammal, the method comprising: a) providing a mammal; b) providing an antibody or antigen-binding fragment thereof as defined herein; c) administering an antibody or antigen-binding fragment thereof as defined herein to a mammal such that the antibody or fragment thereof is capable of binding to the extracellular domain of integrin alpha10 beta1 of cells in said mammal; d) optionally adding a second, labeled antibody or fragment thereof to the sample, said second antibody or fragment thereof binding to the antibody or fragment thereof of c); e) detecting an antibody or antigen-binding fragment thereof as defined herein in c) of said cells, or optionally detecting a second, labeled antibody or fragment thereof in d) bound to the antibody or fragment thereof; f) imaging the detected antibody or fragment thereof, thereby imaging the expression of integrin alpha10 beta1 on mammalian cells in vivo. [Brief description of the drawings]

[0021] [Figure 1] Humanized integrin alpha10 antibody variants inhibit proliferation of C2C12 alpha10 cells. C2C12 alpha10 cells were seeded on 96-well plates coated with collagen type I and treated with five humanized integrin alpha10 antibody variants and human isotype control antibody Th301 (5 μg / ml each). Cell proliferation analyzed by BrdU incorporation is shown relative to proliferation of untreated cells (NT), which was set at 100%.

[0022] [Diagram 2] Humanized integrin alpha10 antibody variants inhibit adhesion of C2C12 alpha10 cells. Cells were pre-incubated with five humanized integrin alpha10 antibody variants and human isotype control Th301 (5 μg / ml each). Cells were allowed to adhere to plates coated with type I collage. Bar graphs show data as mean ± SE of three independent cell adhesion experiments. Cell adhesion is shown relative to adhesion of untreated cells (NT), which was set at 100%.

[0023] [Diagram 3] Humanized integrin alpha10 antibody variants inhibit migration of C2C12 alpha10 cells. Spheroids composed of C2C12 alpha10 cells were embedded in collagen I gel and treated with five humanized integrin alpha10 antibody variants and human isotype control Th301 (10 μg / ml each). Migration of cells from the spheroids was analyzed after 24 hours. Relative migration was quantified as migration area difference. Data are presented as mean ± SE. Cell migration is shown relative to migration of untreated cells (NT), which was set at 100%.

[0024] [Figure 4] High cell binding affinity of TAR-Ab23 compared to mAb365. The figure shows the binding of humanized integrin alpha10 antibody TAR-Ab23 and mouse integrin alpha10 antibody mAb365 to C2C12alpha10 cells (A), triple negative breast cancer cells BT549 (B) and patient-derived glioblastoma U3046MG cells (C) at different concentrations. A comparison of the cell binding ability of the two antibodies at low antibody concentrations (0.1 nM) is presented in figure D. Cells were incubated with the antibodies at the indicated concentrations (nM) followed by incubation with the secondary antibody. Immunolabeled cells were analyzed by flow cytometry.

[0025] [Diagram 5]Higher binding ability of TAR-Ab23 compared to mAb365 in a competition assay. The figure shows the percentage of mean fluorescence intensity (MFI) in a flow cytometry competition assay analyzing the cell binding ability of humanized integrin alpha10 antibody TAR-Ab23 and mouse integrin alpha10 antibody mAb365 to C2C12 alpha10 cells and BT549 cancer cells. Antibodies were added simultaneously at a final concentration of 100 nM. The MFI from single antibody staining was set as 100%.

[0026] [Figure 6] TAR-Ab23 has improved blocking effect on triple-negative breast cancer adhesion, proliferation, and migration compared to mAb365. (A) Adhesion: BT549 cells were treated with 5 μg / ml of human integrin alpha 10 antibody TAR-Ab23 or mouse integrin alpha 10 antibody mAb365 (each at 5 μg / ml) and then allowed to adhere to collagen IV. Cell adhesion is shown relative to adhesion of untreated cells (NT), which was set at 100%. Data represent the mean + / - SD of triplicate measurements. (B) Proliferation: BT549 cancer cells were mixed with 5 μg / ml of TAR-Ab23, mAb365) or control antibodies human Th301 and mouse mIgG2a and plated on collagen IV-coated dishes. After 48 hours, cell proliferation was analyzed by BrdU incorporation. Data are shown relative to proliferation of the respective isotype control antibodies, which was set at 100%. (C) Migration: Spheroids composed of BT549 cells were embedded in collagen I gel and treated with 10 μg / mL of TARG10, mAb365, or control antibodies Th301 and mIgG2a. Cell migration was analyzed after 24 h and quantified as the difference in migration area. Data are presented as the mean ± SE of three independent experiments.

[0027] [Figure 7]TAR-Ab23 inhibits metastasis of TNBC cells. TNBC cells, MDA-MB-231, were labeled with Luc / GFP positive cells and then injected intravenously (2.0×106 cells) into NUDE-NMRI mice. Mice were then treated with 5 mg / kg TAR-Ab23 or PBS as a control. Treatment (twice a week) began 3 days after tumor cell injection and continued for 10 weeks. The graph shows a summary of distant metastasis to organs including lung and liver at the end date. Metastasis was measured using bioluminescence imaging analysis (total flux (P / S)). Results are from 5 mice per group.

[0028] [Figure 8] In vitro ADCC mediated by TAR-Ab23. (A) Induction of ADCC by the Jurkat-Lucia / NFAT-CD16 effector cell line using Tar-Ab23 on A204 as target cells. (B) Induction of ADCC by the Jurkat-Lucia / NFAT-CD16 effector cell line using Tar-Ab23 low fucose on A204 as target cells. The effector / target cell ratio was 2:1 in all cases. Data are presented as relative light units (RLU).

[0029] definition As used herein, the singular forms "a," "an," and "the" include plural references unless the context clearly indicates otherwise. Thus, for example, a reference to an "antibody" includes a plurality of such antibodies. Similarly, an "anti-integrin alpha 10 antibody" can also refer to an "anti-integrin alpha 10 antibody," such as, for example, the antibody variants described in Examples 1-9.

[0030] As used herein, the term "some embodiments" may include one or more than one embodiments.

[0031] As used herein, "integrin alpha10" or "integrin alpha10 subunit" or "integrin alpha10 polypeptide" refers to the alpha10 subunit of the heterodimeric protein integrin alpha10 beta1. This designation does not exclude the presence of a beta1 subunit bound to the alpha10 subunit, thus forming an integrin alpha10 beta1 heterodimer. "Alpha" and "α", as well as "alpha10" and "alpha10" are equivalent terms. As used herein, "integrin alpha10" can also refer to polynucleotide transcripts encoding the alpha10 subunit of the heterodimeric protein integrin alpha10 beta1, and fragments thereof.

[0032] As used herein, "anti-integrin alpha 10 antibody" or "integrin alpha 10 antibody" or "anti-integrin alpha 10 subunit antibody" refers to an antibody capable of recognizing and binding to at least the alpha 10 integrin of the heterodimeric protein integrin alpha 10 beta 1. These antibodies may be antibodies that recognize an epitope of the heterodimeric protein integrin alpha 10 beta 1, where the epitope includes amino acid residues of both the alpha 10 and beta 1 integrin polypeptides.

[0033] As used herein, an "antibody or antigen-binding fragment of the invention" may be referred to as a "polypeptide of the invention" or an "antibody polypeptide, or antigen-binding fragment thereof" since antibodies and fragments thereof are polypeptides.

[0034] As used herein, the term "antibody or antigen-binding fragment thereof" includes substantially intact antibodies, as well as antibody fragments and derivatives. An intact antibody may be considered an antibody comprising a variable light region, a variable heavy region, a constant light region, and a constant heavy region. In addition, it includes chimeric antibodies, humanized antibodies, isolated human antibodies, single chain antibodies, bispecific antibodies, antibody heavy chains, antibody light chains, homodimers and heterodimers of antibody heavy and / or light chains, as well as antigen-binding fragments and derivatives thereof. Suitable antigen-binding fragments and derivatives include, but are not limited to, Fv fragments (e.g., single chain Fvs and disulfide-linked Fvs), Fab-like fragments (e.g., Fab fragments, Fab' fragments, and F(ab)2 fragments), single variable domains (e.g., VH and VL domains), and domain antibodies (dAbs, including single and dual formats [i.e., dAb-linker-dAb]). The potential advantages of using antibody fragments rather than whole antibodies are several-fold. The small size of the fragments may result in improved pharmacological properties, such as increased penetration through solid tissues. Furthermore, antigen-binding fragments, such as Fab, Fv, ScFv and dAb antibody fragments, can be expressed in and secreted from E. coli, allowing for the facile production of large amounts of the fragments.

[0035] As used herein, the term "antibody" or "antigen-binding fragment of the invention" is also intended to encompass antibody mimetics (e.g., non-antibody scaffold structures that have a high degree of stability and allow for the introduction of dispersion at specific locations). Those skilled in the art of biochemistry will be familiar with many such molecules, as discussed in Gebauer & Skerra, 2009, Curr Opin Chem Biol 13(3):245-255, the disclosure of which is incorporated herein by reference. Exemplary antibody mimetics include affibodies (also called trinectins, Nygren, 2008, FEBSJ, 275, 2668-2676); CTLDs (also called tetranectins, Innovations Pharmac, Technol. (2006), 27-30); adnectins (also called monobodies, Meth. Mol. Biol., 352 (2007), 95-109); anticalins (Drug Discovery Today (2005), 10, 23-33); DARPins (ankyrins; Nat. Biotechnol. (2004), 22, 575-582); avimers (Nat. Biotechnol. (2005), 23, 1556-1561); microbodies (FEBS J, (2007), 274, 86-95); peptide aptamers (Expert, Opin, Biol. Ther. (2005), 5, 783-797); Kunitz domains (J. Pharmacol, Exp, Ther. (2006) 318, 803-809); affilins (Trends, Biotechnol. (2005), 23, 514-522); and affimers (Avacta Life Sciences, Wetherby, UK).

[0036] As used herein, the term "antigen-binding fragment" refers to a fragment of an antibody that retains the ability to specifically bind to an antigen. Examples of antibody fragments of the present invention include Fab fragments, Fab' fragments, F(ab')2 fragments, and Fv fragments, such as single chain variable fragments (scFv) and single domain antibodies.

[0037] As used herein, the term "amino acid" includes the standard 20 genetically encoded amino acids and their corresponding "D" stereoisomers (as compared to the natural "L" forms), omega amino acids and other naturally occurring amino acids, unconventional amino acids (e.g., α,α-disubstituted amino acids, N-alkyl amino acids, etc.), and chemically derivatized amino acids as described herein. When an amino acid such as "alanine" or "Ala" or "A" is specifically recited, the term refers to both l-alanine and d-alanine unless otherwise indicated. Other unconventional amino acids may also be suitable components of the polypeptides (antibodies or antigen-binding fragments thereof) of the invention, so long as the desired functional properties are retained by the antibody or antigen-binding fragment. For the amino acid sequences shown, each encoded amino acid residue is represented, where appropriate, by a one-letter designation corresponding to the trivial name of the conventional amino acid.

[0038] As used herein, "expression vector" or "vector" refers to a DNA construct comprising a DNA sequence operably linked to a suitable control sequence capable of effecting expression of the DNA in a suitable host. Such control sequences may include, for example, a promoter for effecting transcription, an optional operator sequence for controlling such transcription, a sequence encoding a suitable mRNA ribosome binding site, and a sequence controlling the termination of transcription and translation. A vector may be, for example, a plasmid, a phage, or simply a potential genome insert. Once transformed into a suitable host, the vector may, for example, replicate and function independently of the host genome, or in some cases, be integrated into the genome itself. Expression vectors are designed, for example, as described in Li et al. (construction strategies for developing expression vectors for recombinant monoclonal antibody production in CHO cells, Mol Biol Rep. 2018 Dec; 45(6): 2907-2912).

[0039] As used herein, "subject" refers to mammals such as rodents, cats, dogs, horses, and primates. Preferably, a subject according to the present invention is a human.

[0040] As used herein, "sample" encompasses any subject and various sample types obtained from any subject. Examples of samples useful in the methods of the present disclosure include, but are not limited to, a subject, a liquid tissue sample such as blood, or a solid tissue sample such as a biopsy or tissue culture or cells derived therefrom and their progeny. For example, a biological sample includes cells obtained from a tissue sample collected from a subject. Thus, samples encompass clinical samples, cells in culture, cell supernatants, cell lysates, and tissue samples, such as tissue samples from breast tissue, lung tissue, prostate tissue, pancreatic tissue, bone tissue, cartilage tissue, adipose tissue, muscle tissue, and connective tissue.

[0041] "Cancer" as used herein refers to any malignant and / or invasive growth or tumor caused by abnormal cell growth. As used herein, "cancer" refers to a tumor named after the type of cells that form the tumor. A cancer or tumor is composed of tumor cells or cancer cells. A cancer or tumor also includes the cancer or tumor microenvironment, which may also include MSCs, fibroblasts, endothelial cells, pericytes, adipocytes, immune cells, tumor-associated macrophages (TAMs). Part of a cancer or tumor may be a stromal cell, e.g., a connective tissue cell, such as a fibroblast. Examples of solid tumors include, but are not limited to, sarcomas and carcinomas. The term "cancer" includes, but is not limited to, primary cancers that occur at a particular site in the body, metastatic cancers that have spread to other parts of the body from where it began, recurrences from the original primary cancer, cancers after remission, and second primary cancers, which are new primary cancers in humans with a history of a previous cancer of a different type than the latter.

[0042] As used herein, "detection," "detect" and "detecting" include qualitative and / or quantitative detection (measuring levels) with or without reference to a control, and more specifically refer to identifying the presence, absence, or amount of a given target that is a target of integrin alpha10 subunit.

[0043] As used herein, "inhibition" means that the presence of the antibody of the present invention inhibits, in whole or in part, the binding of a ligand to the receptor and / or the neutralization of the signal elicited by the receptor upon ligand binding. This includes, for example, downstream signaling that affects cellular behavior and processes. "Inhibition" and "blocking" are used as equivalent terms herein. It should be understood that this inhibition is compared to the situation in the presence of an inhibitor (in the present case, an antibody or antigen-binding fragment thereof having binding specificity for integrin alpha10 beta1) compared to the situation in the absence of the inhibitor. Those skilled in the art will understand that the degree of inhibition can be determined by methods well known in the art depending on the signaling event or pathway or activity being measured.

[0044] As used herein, "inhibition of signal transduction" is defined as a reduction in the signal transduction event, or a reduction in the activity, confirmation, or production of, for example, a receptor or biological pathway or molecular activity. Note that the phrase "inhibit (for example) integrin alpha10 beta1 signal transduction" is used interchangeably with the phrase "inhibit (for example) integrin alpha10 beta1 signal transduction."

[0045] As used herein, "ADCC activity" or "antibody-dependent cellular cytotoxicity" refers to the activity of damaging target cells (e.g., tumor cells) by activating effector cells through binding of the Fc region of an antibody to Fc receptors present on the surface of effector cells such as killer cells, natural killer cells, and activated macrophages. The activity of the antibody of the present invention includes ADCC activity. ADCC activity measurement and anti-tumor experiments can be performed using any assay known in the art.

[0046] As used herein, "pharmaceutical acceptable" refers to a non-toxic material that does not reduce the effectiveness of the integrin alpha10 beta1 binding activity of the antibody or antigen-binding fragment of the present invention. Pharmaceutically acceptable buffers, carriers, diluents, or excipients are well known in the art.

[0047] "Treatment" includes both therapeutic and prophylactic or preventive treatment of a patient. The terms "preventive" or "prophylactic" are used to encompass the use of an antibody or antigen-binding fragment thereof, or a formulation thereof, described herein, to either prevent or reduce the likelihood of a neoplastic disorder or disorder, including preventing or reducing the spread, dissemination, or metastasis of neoplastic cells in a patient or subject. The term "prophylactic" also encompasses the use of an antibody or antigen-binding fragment thereof, or a formulation thereof, described herein, to prevent the recurrence of a neoplastic disease or disorder in a patient who has previously been treated for either of these. "Treatment" and "palliative" are used interchangeably herein.

[0048] By "diagnosis" we include the detection of cells associated with a neoplastic disease or disorder either in vivo (i.e., within a patient's body) or ex vivo (i.e., in a tissue or cell sample removed from a patient's body).

[0049] By "disorders associated with cells expressing integrin alpha10 beta1", we include diseases or disorders in which pathological cells that directly or indirectly cause the disorder express integrin alpha10 beta1 on the cell surface. It will be understood that cells expressing integrin alpha10 beta1 can be immune cells, fibroblasts or cells of connective tissue such as tumor cells (cancer cells), e.g., tumor cells themselves. Furthermore, such cells include pathological stem cells (i.e., cancer stem cells, or CSCs) and progenitor cells that are directly or indirectly involved in the development of neoplastic diseases or disorders in individuals. Examples of CSCs are disclosed in Visvader & Lindeman, 2008, Nat Rev Cancer 8:755-768, the disclosure of which is incorporated herein by reference.

[0050] Alternatively, or in addition, cells expressing integrin alpha10beta1 may be indirectly associated with a neoplastic disease or disorder, for example they may mediate cellular processes necessary for survival.

[0051] "Buffer" as used herein refers to an aqueous solution containing an acid-base mixture for the purpose of stabilizing pH. Examples of buffers are trizma, bicine, tricine, MOPS, MOPSO, MOBS, tris, Hepes, HEPBS, MES, phosphate, carbonate, acetate, citrate, glycolate, lactate, borate, ACES, ADA, tartrate, AMP, AMPD, AMPSO, BES, CABS, cacodylate, CHES, DIPSO, EPPS, ethanolamine, glycine, HEPPSO, imidazole, imidazole lactate, PIPES, SSC, SSPE, POPSO, TAPS, TABS, TAPSO and TES.

[0052] As used herein, "diluent" refers to an aqueous or non-aqueous solution for the purpose of diluting the antibody or antigen-binding fragment in a pharmaceutical formulation. The diluent may be one or more of saline, water, polyethylene glycol, propylene glycol, ethanol, or oil (such as safflower oil, corn oil, peanut oil, cottonseed oil, or sesame oil).

[0053] As used herein, "adjuvant" refers to any substance that, when mixed with an administered antigen, increases or otherwise modulates the immune response to the antigen. Suitable adjuvants are well known to those skilled in the art.

[0054] As used herein, "carrier" refers to a scaffold structure, such as a polypeptide or polysaccharide, to which an antigen can be associated. A carrier can exist independently of an adjuvant. Suitable carriers are well known by those skilled in the art.

[0055] As used herein, "therapeutically effective amount" or "effective amount" or "therapeutically effective" refers to an amount that produces a therapeutic effect for a given condition and administration regimen. It is a predetermined amount of an active agent calculated to produce a desired therapeutic effect in association with the necessary additives and diluents, i.e., carriers or administration vehicles. Moreover, it is intended to mean an amount sufficient to reduce, and most preferably prevent, a clinically significant deficiency in the activity, function and response of the host. Alternatively, a therapeutically effective amount is sufficient to improve a clinically significant condition in the host. As will be appreciated by those skilled in the art, the amount of a compound may vary depending on its specific activity. A suitable dosage may include a predetermined amount of an active composition calculated to produce a desired therapeutic effect in association with the necessary diluents. In the method of making and using the composition of the present invention, a therapeutically effective amount of the active ingredient is provided. A therapeutically effective amount can be determined by a physician or veterinarian of ordinary skill based on the characteristics of the patient, such as age, weight, sex, condition, comorbidities, other diseases, etc., as is well known in the art. The administration of a pharma- ceutically effective dose can be carried out by a single administration in the form of an individual or several smaller dose units, or by multiple administrations at specific intervals of subdivided doses, or the dose can be provided as a continuous infusion over an extended period of time. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0056] Integrin alpha 10 antibody In one embodiment, the present invention relates to an antibody or antigen-binding fragment thereof that has binding specificity for integrin alpha10 beta1, the antibody or antigen-binding fragment being: A light chain variable region, a) a CDR-L1 comprising or consisting of the amino acid sequence of SEQ ID NO: 1; b) a CDR-L2 comprising or consisting of an amino acid sequence selected from the group consisting of SEQ ID NO: 2 and SEQ ID NO: 4; and c) a light chain variable region comprising a CDR-L3 comprising or consisting of the amino acid sequence of SEQ ID NO:5; and / or A heavy chain variable region, a) a CDR-H1 comprising or consisting of an amino acid sequence selected from the group consisting of SEQ ID NO: 7, SEQ ID NO: 6 and SEQ ID NO: 8; b) a CDR-H2 comprising or consisting of the amino acid sequence of SEQ ID NO: 9; and c) a heavy chain variable region comprising a CDR-H3 comprising or consisting of the amino acid sequence of SEQ ID NO:10.

[0057] In some embodiments, the antibody or antigen-binding fragment thereof comprises: A light chain variable region, a) CDR-L1 consisting of the amino acid sequence of SEQ ID NO: 1; b) a CDR-L2 consisting of an amino acid sequence selected from the group consisting of SEQ ID NO: 2 and SEQ ID NO: 4; and c) a light chain variable region comprising a CDR-L3 consisting of the amino acid sequence of SEQ ID NO:5; and / or A heavy chain variable region, a) CDR-H1 consisting of an amino acid sequence selected from the group consisting of SEQ ID NO: 7, SEQ ID NO: 6 and SEQ ID NO: 8; b) CDR-H2 consisting of the amino acid sequence of SEQ ID NO: 9, and c) A heavy chain variable region comprising a CDR-H3 consisting of the amino acid sequence of SEQ ID NO:10.

[0058] The antibody or antigen-binding fragment of the present invention has specificity for the alpha10 subunit in integrin alpha10 beta1. By "specificity" is meant that the antibody or antigen-binding fragment is capable of binding to integrin alpha10 beta1 in vivo, i.e. under physiological conditions in which integrin alpha10 beta1 exists in the human body. Preferably, the antibody or antigen-binding fragment does not bind or has only weak binding to other proteins in vivo. Alternatively, it means that the antibody or antigen-binding fragment is capable of binding to integrin alpha10 beta1 ex vivo or in vitro. Such binding specificity can be confirmed by methods well known in the art, such as ELISA, immunohistochemistry, immunoprecipitation, Western blot, and flow cytometry, using transfected cells expressing integrin alpha10 beta1. Advantageously, the antibody or antigen-binding fragment is capable of selectively binding to integrin alpha10 beta1, i.e. it binds to integrin alpha10 beta1 at least 10 times stronger than any other protein.

[0059] In some embodiments, the antibody or antigen-binding fragment thereof that has binding specificity for integrin alpha10 beta1 is humanized.

[0060] In some embodiments, the antibody or antigen-binding fragment thereof that has binding specificity for integrin alpha10 is a human antibody.

[0061] In some embodiments, the antibody or antigen-binding fragment thereof that has binding specificity for integrin alpha10 is deimmunized.

[0062] In some embodiments, the antibody or antigen-binding fragment thereof that has binding specificity for integrin alpha10 is humanized and deimmunized.

[0063] In some embodiments, the antibody or antigen-binding fragment that has binding specificity for integrin alpha10 binds to human integrin alpha10 beta1.

[0064] In some embodiments, the antibody or antigen-binding fragment thereof that has binding specificity for integrin alpha10 binds to a non-human integrin alpha10 beta1, for example, cynomolgus monkey.

[0065] In some embodiments, the antibody or antigen-binding fragment thereof that has binding specificity for integrin alpha10 binds to integrin alpha10 beta1 expressed on the surface of a cell.

[0066] In some embodiments, the antibody or antigen-binding fragment thereof that has binding specificity for integrin alpha10 binds to an epitope on the extracellular domain I of subunit alpha10 of integrin alpha10 beta1.

[0067] In some embodiments, the antibody or antigen-binding fragment thereof that has binding specificity for integrin alpha10 beta1 binds to the extracellular domain I of subunit alpha10 of integrin alpha10 beta1.

[0068] In some embodiments, as described above, the antibody or antigen-binding fragment of the present invention comprises or consists of an antibody mimetic selected from the group comprising or consisting of affibodies, tetranectins (CTLDs), adnectins (monobodies), anticalins, DARPins (ankyrins), avimers, iMabs, microbodies, peptide aptamers, Kunitz domains, and affilins.

[0069] Those skilled in the art will further appreciate that the present invention also encompasses modified versions of antibodies and antigen-binding fragments thereof, whether existing now or in the future, such as those modified by the covalent attachment of polyethylene glycol or another suitable polymer (see below).

[0070] Integrin alpha 10 polypeptide Integrins are heterodimers consisting of alpha and beta polypeptides. Integrin alpha10 beta1 heterodimers can be detected by integrin alpha10 specific antibodies, as well as integrin alpha10 binding peptides and proteins.

[0071] In some embodiments, the integrin alpha10 polypeptide is part of an integrin alpha10 beta1 heterodimer.

[0072] In some embodiments, the integrin alphal0 polypeptide is expressed on the surface of the cell.

[0073] Integrin alpha10 beta1 was first identified in 1998 as a collagen type II binding receptor on chondrocytes (Camper et al., 1998). In vitro studies have demonstrated binding to other collagen subtypes and laminin (Lundgren-Akerlund Book chapter and Thoren et al). Immunohistochemical analysis in developing and adult tissues demonstrated restricted localization to cartilage-containing tissues and some fibrous tissues (Camper et al. 1998, Camper et al., 2001). Knockout mice lacking this marker have disorganized growth plates, reduced collagen in the matrix, and shortened long bones, further supporting the structural importance of the cell (Bengtsson et al., 2005). The amino acid sequence, variants, isoforms, and sequence annotation can be found in Uniprot accession number O75578 (ITA10_HUMAN).

[0074] Integrin alpha10 beta1 receptor, upon binding to extracellular ligands, transmits intracellular signals that promote cell adhesion, migration, survival, proliferation, tumor growth and metastasis. Inhibition of the receptor thus prevents adhesion, migration, survival, proliferation, tumor growth and metastasis. This may be important for the treatment of many proliferative diseases, such as cancer and inflammatory diseases.

[0075] In some embodiments, the integrin alpha10 is a naturally occurring variant of an integrin alpha10 polypeptide, an isoform of an integrin alpha10 polypeptide, or a splice variant of an integrin alpha10 polypeptide.

[0076] Integrin alpha10 can also be detected at the nucleotide level, for example, by analyzing a sample for the presence of mRNA transcripts which, upon translation, generate the integrin alpha10 antigen as defined herein above.

[0077] CDR The antibodies of the present invention are defined by their characteristic complementarity determining region (CDR) sequences. There are several approaches to define the CDR sequences of an antibody. The CDRs of the antibodies of the present invention are defined using the definition according to Kabat.

[0078] Those skilled in the art will understand that the set of six CDRs (CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2, CDR-H3) may be defined according to Kabat.

[0079] Furthermore, those skilled in the art will understand that it is possible to define the CDRs of the antibodies of the present invention by other approaches, for example by the CDR definitions according to Chothia (Al-Lazikani et al., (1997) JMB 273, 927-948), Martin (Enhanced Chotia), Gelfand, or Honneger. Further approaches exist, such as the AbM definition (a combination of the Kabat and Chothia definitions used by Oxford Molecular's AbM antibody modeling software) or the contact definition (based on the analysis of crystal structures). See, e.g., Kabat et al. (Sequences of Proteins of Immunological Interest, 1987 and 1991, NIH, Bethesda, Md.), Lefranc et al. (IMGT unique numbering for immunoglobulin and T cell receptor constant domains and Ig superfamily C-like domains, Dev Comp Immunol. 2005;29(3):185-203) and Dondelinger et al. (Understanding the Significance and Implications of Antibody Numbering and Antigen-Binding Surface / Residue Definition, Front, Immunol., 16 October 2018).

[0080] Those skilled in the art can use well-known information to enumerate other CDR naming conventions or approaches (such as Chothia) when given the Kabat CDRs presented herein. Thus, all CDR naming conventions or approaches are encompassed.

[0081] In certain cases, it may be useful to define CDR according to one numbering system, such as Kabat.In many cases, these CDR sequences are short (e.g., shorter than the approach of combining numbering systems), and therefore provide important core sequences for binding.In other cases, it may be useful to use, for example, a combination of IMGT sequence and Kabat CDR sequence.

[0082] However, one of skill in the art will understand that low levels of mutations within the CDR sequences (typically only one or two amino acids) can be tolerated without loss of specificity of the antibody or antigen-binding fragment for integrin alpha10.

[0083] In some embodiments, the antibody or antigen-binding fragment thereof that has binding specificity for integrin alpha 10 comprises a light chain variable region comprising: a) a CDR-L1 comprising or consisting of the amino acid sequence of SEQ ID NO: 1; b) a CDR-L2 comprising or consisting of an amino acid sequence selected from the group consisting of SEQ ID NO:2; and c) a light chain variable region comprising a CDR-L3 comprising or consisting of the amino acid sequence of SEQ ID NO:5; And, A heavy chain variable region, a) a CDR-H1 comprising or consisting of an amino acid sequence selected from the group consisting of SEQ ID NO: 7; b) a CDR-H2 comprising or consisting of the amino acid sequence of SEQ ID NO: 9; and c) a heavy chain variable region comprising a CDR-H3 comprising or consisting of the amino acid sequence of SEQ ID NO: 10; Or, it includes an amino acid sequence having at least 70% sequence identity, for example at least 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:5, SEQ ID NO:7, or SEQ ID NO:9, SEQ ID NO:10.

[0084] In some embodiments, an antibody or antigen-binding fragment thereof with binding specificity for integrin alpha 10 comprises the above-described CDRs (comprising or consisting of an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-10), in which any one of the amino acids in the CDR is modified for another amino acid, provided that no more than two amino acids are so modified, e.g., one amino acid.

[0085] Light chain variable region In some embodiments, the antibody or antigen-binding fragment thereof that has binding specificity for integrin alpha 10 is a) an amino acid sequence of SEQ ID NO: 13 or an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 13, such as at least 90%, 95%, 98%, or 99% sequence identity; or b) comprising a light chain variable region comprising, or consisting of, an amino acid sequence selected from the group consisting of SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:14, and SEQ ID NO:15, or an amino acid sequence having at least 85% sequence identity to SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:14, and SEQ ID NO:15, such as at least 90%, 95%, 98%, or 99% sequence identity to any one of SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:14, and SEQ ID NO:15.

[0086] In some embodiments, an antibody or antigen-binding fragment thereof with binding specificity for integrin alpha10 comprises a light chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO:13, or an amino acid sequence having at least 85% sequence identity, e.g., at least 90%, 95%, 98% or 99% sequence identity to SEQ ID NO:13.

[0087] Percentage of identity (or sequence identity) can be determined, for example, by the LALIGN program at the Expasy site (http: / / www.ch.embnet.org / software / LALIGN_format.html) using the following parameters: global alignment option, score matrix BLOSUM62, opening gap penalty -14, extension gap penalty -4. Alternatively, the percentage of sequence identity between two polypeptides, such as portions of an antibody, may be determined using a suitable computer program, for example the GAP program of the University of Wisconsin Genetic Computing Group, it being understood that the percentage of identity is calculated for the polypeptides whose sequences are optimally aligned.

[0088] The alignment may alternatively be performed using the Clustal W program. The parameters used may be as follows: -Fast pairwise alignment parameters: K tuple (word) size; 1, window size; 5, gap penalty; 3, number of upper diagonals; 5. Scoring method: x percent. -Multiple alignment parameters: Gap open penalty; 10, Gap extension penalty; 0.05. -Score matrix: BLOSUM.

[0089] Alternatively, the BESTFIT program may be used to determine local sequence alignments.

[0090] One skilled in the art would consider further modifications to the light chain variable region described above, for example to further optimize the antibody or antigen-binding fragment. Typically, one skilled in the art would consider modifying amino acids within the framework regions, i.e., outside the epitope-binding CDR regions, as is done during humanization and deimmunization procedures, thereby not typically modifying the CDR regions.

[0091] In some embodiments, an antibody or antigen-binding fragment thereof with binding specificity for integrin alpha10 comprises a light chain variable region as described above, wherein any one of the amino acids in the framework regions of the light chain variable region is altered to another amino acid, provided that no more than five amino acids are altered, such as four amino acids, no more than three amino acids, e.g., no more than two amino acids, or no more than one amino acid.

[0092] Heavy chain variable region In some embodiments, the antibody or antigen-binding fragment thereof that has binding specificity for integrin alpha 10 is a) an amino acid sequence of SEQ ID NO: 19 or an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 19, such as at least 90%, 95%, 98%, or 99% sequence identity; or b) comprising a heavy chain variable region comprising, or consisting of, an amino acid sequence selected from the group consisting of SEQ ID NO:16, SEQ ID NO:17, SEQ ID NO:18, and SEQ ID NO:20, or an amino acid sequence having at least 85% sequence identity to SEQ ID NO:16, SEQ ID NO:17, SEQ ID NO:18, and SEQ ID NO:20, such as at least 90%, 95%, 98%, or 99% sequence identity to any one of SEQ ID NO:16, SEQ ID NO:17, SEQ ID NO:18, and SEQ ID NO:20.

[0093] In some embodiments, an antibody or antigen-binding fragment thereof with binding specificity for integrin alpha10 comprises a heavy chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO:19 or an amino acid sequence having at least 85% sequence identity to SEQ ID NO:19, e.g., at least 90%, 95%, 98% or 99% sequence identity.

[0094] One skilled in the art would consider further modifications to the heavy chain variable regions described above, for example to further optimize the antibody or antigen-binding fragment. Typically, one skilled in the art would consider modifying amino acids within the framework regions, i.e., outside the epitope-binding CDR regions, and thereby not modifying the CDR regions, as is done during humanization and deimmunization procedures.

[0095] In some embodiments, an antibody or antigen-binding fragment thereof with binding specificity for integrin alpha10 beta1 comprises a heavy chain variable region as described above, wherein any one of the amino acids in the framework regions of the heavy chain variable region is altered to another amino acid, provided that no more than five amino acids are altered, such as four amino acids, no more than three amino acids, e.g., no more than two amino acids, or no more than one amino acid.

[0096] Variable light chain and variable heavy chain combinations It will be appreciated by those skilled in the art that any of the light chain variable region variants described above can be combined with any of the heavy chain variable region variants described above.

[0097] In some embodiments, an antibody or antigen-binding fragment thereof that has binding specificity for integrin alpha 10 comprises a light chain variable region that comprises, or consists of, SEQ ID NO: 13, and a heavy chain variable region that comprises, or consists of SEQ ID NO: 19; Or, it comprises an amino acid sequence having at least 70% sequence identity to SEQ ID NO:13 or SEQ ID NO:19, such as at least 80%, 85%, 90%, 95%, 98%, or 99% sequence identity.

[0098] In some embodiments, the antibody or antigen-binding fragment thereof that has binding specificity for integrin alpha 10 is a) a light chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO: 11, and a heavy chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO: 16; b) a light chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO: 11, and a heavy chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO: 17; c) a light chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO: 11, and a heavy chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO: 18; d) a light chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO: 11, and a heavy chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO: 19; e) a light chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO: 11, and a heavy chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO: 20; f) a light chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO: 12, and a heavy chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO: 16; g) a light chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO: 12, and a heavy chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO: 17; h) a light chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO: 12, and a heavy chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO: 18; i) a light chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO: 12, and a heavy chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO: 19; j) a light chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO: 12, and a heavy chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO: 20; k) a light chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO: 13, and a heavy chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO: 16; l) a light chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO: 13, and a heavy chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO: 17; m) a light chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO: 13, and a heavy chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO: 18; n) a light chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO: 13, and a heavy chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO: 19; o) a light chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO: 13, and a heavy chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO: 20; p) a light chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO: 14, and a heavy chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO: 16; q) a light chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO: 14, and a heavy chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO: 17; r) a light chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO: 14, and a heavy chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO: 18; s) a light chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO: 14, and a heavy chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO: 19; t) a light chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO: 14, and a heavy chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO: 20; u) a light chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO: 15, and a heavy chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO: 16; v) a light chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO: 15, and a heavy chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO: 17; w) a light chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO: 15, and a heavy chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO: 18; x) a light chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO: 15, and a heavy chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO: 19; y) a light chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO: 15, and a heavy chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO: 20; Alternatively, it comprises an amino acid sequence having at least 70% sequence identity, for example at least 80%, 85%, 90%, 95%, 98%, or 99% sequence identity to any one of SEQ ID NOs: 11 to 20.

[0099] These combinations of light and heavy chain variable regions are, for example, part of the humanized antibody variants described in Examples 1 and 2.

[0100] In some embodiments, an antibody or antigen-binding fragment thereof with binding specificity for integrin alpha10 comprises a heavy chain variable region as described above, in which any one of the amino acids in the framework regions of the light chain variable region and / or the heavy chain variable region is altered to another amino acid, provided that no more than five amino acids, such as four amino acids, no more than three amino acids, e.g., no more than two amino acids, or no more than one amino acid, are altered.

[0101] Constant regions and Fc portions A person skilled in the art will appreciate that any light constant region known in the art can be combined with any of the above light variable region variants, and any heavy constant region known in the art can be combined with any of the above heavy variable region variants to form a complete antibody.

[0102] In some embodiments, the antibody or antigen-binding fragment thereof that has binding specificity for integrin alphal0 comprises a light chain constant region or a portion thereof.

[0103] In some embodiments, the antibody or antigen-binding fragment thereof that has binding specificity for integrin alpha10 comprises a light chain constant region that is a kappa light chain or a lambda light chain.

[0104] In some embodiments, the antibody or antigen-binding fragment thereof that has binding specificity for integrin alphal0 comprises a heavy chain constant region or a portion thereof.

[0105] In some embodiments, the antibody or antigen-binding fragment thereof that has binding specificity for integrin alphal0 comprises a heavy chain constant region selected from the group consisting of α, δ, γ, ε, and μ.

[0106] In some embodiments, the antibody or antigen-binding fragment thereof that has binding specificity for integrin alpha10 comprises a heavy chain constant region that is a human immunoglobulin isotype selected from the group consisting of IgA, IgD, IgG, IgE, and IgM.

[0107] In some embodiments, the antibody or antigen-binding fragment thereof that has binding specificity for integrin alphal0 comprises a heavy chain constant region that is an IgG human immunoglobulin isotype.

[0108] In some embodiments, the antibody or antigen-binding fragment thereof that has binding specificity for integrin alpha10 is an IgG antibody.

[0109] In some embodiments, the antibody or antigen-binding fragment thereof that has binding specificity for integrin alpha10 comprises a heavy chain constant region that is a human immunoglobulin subclass selected from the group consisting of IgG1, IgG2, IgG3, and IgG4.

[0110] In some embodiments, an antibody or antigen-binding fragment thereof with binding specificity for integrin alpha10 comprises or consists of a light chain constant region and / or a heavy chain constant region, in which any one of the amino acids in said light chain constant region and / or said heavy chain constant region is altered to another amino acid, provided that no more than five amino acids are altered, such as four amino acids, no more than three amino acids, e.g., no more than two amino acids, or no more than one amino acid.

[0111] In some embodiments, the antibody or antigen-binding fragment of the invention comprises the CH1, CH2 and / or CH3 regions of an IgG heavy chain (such as an IgG1, IgG2, IgG3 or IgG4 heavy chain). Thus, the antibody or antigen-binding fragment may comprise part or all of the constant region from an IgG1 heavy chain. For example, the antibody or antigen-binding fragment may be a Fab fragment comprising the CH1 and CL constant regions in combination with any of the heavy and light chain variable regions, respectively, defined above.

[0112] Similarly, the above-defined antibody or antigen-binding fragment of the present invention may further comprise a light chain constant region or a part thereof. For example, the antibody or antigen-binding fragment may comprise a CL region from a kappa or lambda light chain.

[0113] In some embodiments, the antibody or antigen-binding fragment thereof that has binding specificity for integrin alpha10 beta1 is any one of the light chain variable regions listed above, and / or any one of the heavy chain variable regions listed above, and / or - any one of the light chain constant regions listed above, and / or - comprising or consisting of any one of the heavy chain constant regions above.

[0114] In some embodiments, the antibody or antigen-binding fragment thereof that has binding specificity for integrin alphal0 comprises an Fc region. The Fc region may also be referred to as an Fc domain.

[0115] In some embodiments, the antibody or antigen-binding fragment thereof that has binding specificity for integrin alpha10 comprises a native Fc region.

[0116] In some embodiments, the antibody or antigen-binding fragment thereof that has binding specificity for integrin alpha10 comprises a non-native Fc region.

[0117] In some embodiments, the antibody or antigen-binding fragment thereof that has binding specificity for integrin alphal0 comprises an Fc region that has an altered, eg, mutated, IgG constant region.

[0118] The Fc region may be naturally occurring (eg, part of an endogenously produced antibody) or may be artificial (eg, containing one or more point mutations relative to a naturally occurring Fc region).

[0119] As is well documented in the art, the Fc region of an antibody mediates its serum half-life and effector functions, such as complement-dependent cytotoxicity (CDC), antibody-dependent cellular cytotoxicity (ADCC), and antibody-dependent cellular phagocytosis (ADCP).

[0120] Engineering the Fc region of therapeutic monoclonal antibodies or Fc fusion proteins allows the generation of molecules that are more suited to their required pharmacological activity (Strohl, 2009, Curr Opin Biotechnol 20(6):685-91, the disclosure of which is incorporated herein by reference). Improved Fc binding can also be obtained by applying specific culture conditions, e.g., low fucose conditions.

[0121] Inhibition of biological activity In some embodiments, the integrin alphal0 antibody of the invention is an antibody capable of inhibiting the biological and functional activity of integrin alphal0beta1.

[0122] In some embodiments, an antibody or antigen-binding fragment thereof having binding specificity for integrin alpha 10 can inhibit signaling of integrin alpha 10 beta 1. One skilled in the art will appreciate that inhibition occurs when the antibody or antigen-binding fragment binds to the antibody epitope on integrin alpha 10. Integrin alpha 10 beta 1 is involved in both outside-in and inside-out signaling, and both signaling can be inhibited by using the antibodies of the present disclosure.

[0123] In some embodiments, an antibody or antigen-binding fragment thereof that has binding specificity for integrin alpha10 is capable of inhibiting signal transduction that occurs upon binding to integrin alpha10.

[0124] In some embodiments, an antibody or antigen-binding fragment thereof having binding specificity for integrin alpha10 can inhibit cell survival, cell adhesion, cell proliferation, cell growth, cell migration, or any combination thereof, of cells expressing integrin alpha10 beta 1. One skilled in the art will appreciate that inhibition occurs when the antibody or antigen-binding fragment binds to an epitope on integrin alpha10.

[0125] In some embodiments, an antibody or antigen-binding fragment thereof that has binding specificity for integrin alpha10 is capable of inhibiting cell survival of cells expressing integrin alpha10 beta1.

[0126] In some embodiments, an antibody or antigen-binding fragment thereof that has binding specificity for integrin alpha10 is capable of inhibiting cell adhesion of cells expressing integrin alpha10 beta1.

[0127] In some embodiments, an antibody or antigen-binding fragment thereof that has binding specificity for integrin alpha10 is capable of inhibiting cell proliferation of cells expressing integrin alpha10 beta1.

[0128] In some embodiments, an antibody or antigen-binding fragment thereof that has binding specificity for integrin alpha10 is capable of inhibiting cell growth of cells that express integrin alpha10 beta1.

[0129] In some embodiments, an antibody or antigen-binding fragment thereof that has binding specificity for integrin alpha10 is capable of inhibiting cell migration of cells expressing integrin alpha10 beta1.

[0130] In some embodiments, an antibody or antigen-binding fragment thereof having binding specificity for integrin alpha10 can inhibit cell metastasis of cells expressing integrin alpha10 beta 1. One skilled in the art will appreciate that inhibition occurs when the antibody or antigen-binding fragment binds to an epitope on integrin alpha10.

[0131] In some embodiments, an antibody or antigen-binding fragment thereof having binding specificity for integrin alpha10 can inhibit tumor growth of tumors comprising cells that express integrin alpha10 in the tumor itself and / or in the tumor microenvironment.

[0132] Those skilled in the art will appreciate that the binding of an antibody or antigen-binding fragment thereof to integrin alpha10 beta1 can affect integrin alpha10 beta1 in various ways, for example at the direct molecular level and / or at the conformational level. For example, the binding of an antibody or antigen-binding fragment thereof to integrin alpha10 beta1 can block ligand binding and thus alter signal transduction of integrin alpha10 beta1. It can also result in internalization of the antibody or antigen-binding fragment thereof.

[0133] One skilled in the art will appreciate that not all integrin alphal0 binding antibodies that bind to any one epitope of integrin alphal0 beta 1 may inhibit cell survival, cell adhesion, cell proliferation, cell growth, cell migration, metastasis, tumor growth, integrin alphal0 beta 1 signaling and / or any combination thereof. It is an accomplishment of the present invention to provide such antibodies.

[0134] Those skilled in the art will understand that there are various degrees of inhibition of biological activities, such as cell survival, cell adhesion, cell proliferation, cell growth, cell migration, metastasis, tumor growth, integrin alpha10 beta1 signaling, and / or any combination thereof. Inhibition can be complete or essentially complete. Inhibition can also be partial, e.g., reduced, e.g., non-complete. "Essentially complete" should be understood as "complete" in consideration of the uncertainty of assessing completeness associated with the methodology used to measure inhibition of biological activity.

[0135] For example, biological activity such as cell survival, cell adhesion, cell proliferation, cell growth, cell migration, tumor growth, metastasis, integrin alpha10 beta1 signaling, and / or any combination thereof may be inhibited by at least 10%, 20%, 30%, 50%, 60%, 75%, 80%, 85%, 90%, 95%, 98%, 99% or more compared to biological activity in the absence of an antibody or antigen-binding fragment of the invention.

[0136] In some embodiments, inhibition of biological activity, such as cell survival, cell adhesion, cell proliferation, cell growth, cell migration, tumor growth, metastasis, integrin alpha10 beta1 signaling, and / or any combination thereof, is 10-100% compared to biological activity in the absence of an antibody or antigen-binding fragment of the invention. More preferably, inhibition of biological activity is 25-100%. Even more preferably, inhibition of biological activity is 50-100%.

[0137] The extent of inhibition of biological activities such as cell survival, cell adhesion, cell proliferation, cell growth, cell migration, tumor growth, metastasis, integrin alpha10 beta1 signaling and / or any combination thereof by the antibodies or antigen-binding fragments of the invention can be determined using methods well known in the art, such as the methods used in Examples 7 and 8.

[0138] In a preferred embodiment, an antibody or antigen-binding fragment thereof that has binding specificity for integrin alpha10 is capable of essentially completely inhibiting metastasis.

[0139] The antibodies may also inhibit metastasis to some extent, ie, partially rather than essentially completely.

[0140] In some embodiments, an antibody or antigen-binding fragment thereof that has binding specificity for integrin alpha10 beta1 can partially inhibit metastasis.

[0141] Those skilled in the art will understand that integrin alpha10 antibodies, apart from the above-specified properties, may exhibit one or more additional functions. These functions may be conveyed by the constant region or Fc region of the antibody. Examples of these functions are antibody-dependent cell-mediated cytotoxicity (ADCC) and / or antibody-dependent cellular phagocytosis (ADCP) and / or complement-dependent cytotoxicity (CDC), thereby resulting in the killing of target cells, such as integrin alpha10 beta1-expressing tumor cells.

[0142] In some embodiments, an antibody or antigen-binding fragment thereof that has binding specificity for integrin alpha10 is capable of inducing ADCC of cells expressing integrin alpha10 beta1.

[0143] In some embodiments the antibody or antigen-binding fragment thereof that has binding specificity for integrin alpha10 is a low fucose variant and is capable of inducing ADCC of cells expressing integrin alpha10 beta1.

[0144] In some embodiments, an antibody or antigen-binding fragment thereof that has binding specificity for integrin alpha10 is capable of inducing ADCP of cells expressing integrin alpha10 beta1.

[0145] In some embodiments, an antibody or antigen-binding fragment thereof that has binding specificity for integrin alpha10 is capable of inducing CDC of cells expressing integrin alpha10 beta1.

[0146] As described in detail in the description below, antibodies can be subject to modification to optimize particular properties.

[0147] Antibody Modifications In some embodiments, the antibody or antigen-binding fragment thereof that has binding specificity for integrin alphal0 further comprises a moiety to increase the in vivo half-life of the agent.

[0148] In some embodiments, the moiety for increasing the in vivo half-life of an antibody or antigen-binding fragment thereof having binding specificity for integrin alpha10 is selected from the group consisting of polyethylene glycol (PEG), human serum albumin, glycosylation groups, fatty acids, and dextran.

[0149] In some embodiments, the antibody or antigen-binding fragment thereof that has binding specificity for integrin alpha10 is pegylated.

[0150] In some embodiments, an antibody or antigen-binding fragment thereof that has binding specificity for an integrin alphal0 fragment is directly or indirectly covalently linked to a functional moiety, such as a cytotoxic moiety or a detectable moiety.

[0151] In some embodiments, the antibody or antigen-binding fragment thereof that has binding specificity for integrin alpha10 comprises a cytotoxic moiety.

[0152] In some embodiments, the cytotoxic moiety comprises or consists of a radioisotope.

[0153] In some embodiments, the radioisotope is selected from the group consisting of beta emitters, auger emitters, conversion electron emitters, alpha emitters, and low photon energy emitters.

[0154] In some embodiments, the radioisotope has a locally absorbed energy emission pattern that results in a high dose of absorbance in the vicinity of the agent.

[0155] In some embodiments, the radioisotope is 90 Y, 32 P, 186 Re / 186 Re; 166 Ho, 76 As / 77 As, 153 Sm; 131 I, 177 Lu, 67 Cu, 161 mid-range beta emitters such as Tb; 45 Ca, 35 S, or 14 low energy beta emitters such as C; 51 Cr, 67 Ga, 99 Tc m , 111 In, 123 I, 125 I, 201 Transformation or Auger emitters such as Tl; 212 Bi, 213 Bi, 223 Ac, and 221 At is selected from the group consisting of alpha emitters such as At.

[0156] In some embodiments, the antibody or antigen-binding fragment thereof that has binding specificity for integrin alpha10 beta1 comprises a cytotoxic moiety that comprises or consists of a cytotoxic agent.

[0157] In some embodiments, the cytotoxic drug is selected from the group consisting of cytostatic drugs; antiandrogens; cortisone and its derivatives; phosphates; testosterone-5-α-reductase inhibitors; boron additives; cytokines; thapsigargin and its metabolites; toxins (such as saporin or calicheamicin); chemotherapeutic drugs (such as antimetabolites); or any other cytotoxic drug useful in the treatment of neoplastic disorders.

[0158] In some embodiments, the neoplastic disease or disorder is any of the neoplastic diseases or disorders defined in detail in the "Clinical Conditions" section herein.

[0159] In some embodiments, the cytotoxic moiety is selected from the group consisting of a toxin, a chemotherapeutic agent, and a radioactive agent, or a combination thereof.

[0160] In some embodiments, the toxin is selected from the group selected from a microtubule toxin, a DNA toxin, and a transcription toxin.

[0161] In some embodiments, the microtubule toxin is selected from the group consisting of an auristatin toxin, a maytansinoid toxin, a tubulysin toxin, and an eribulin toxin.

[0162] In some embodiments, the cytotoxic moiety is a DNA toxin selected from the group consisting of DNA minor groove binders, DNA minor groove binding alkylating agents, DNA alkylating agents, and DNA cleaving agents. For example, in some embodiments, the cytotoxic moiety is a DNA toxin selected from the group consisting of pyrrolobenzodiazepines (PBDs), duocarmycins, duocarmycin analogs, indolino-benzodiazepines, calicheamicins, irinotecans, and exatecan derivatives.

[0163] In some embodiments, the transcription toxin is an RNA polymerase II inhibitor.

[0164] In some embodiments, the transcriptional toxin is selected from the group consisting of doxorubicin, a doxorubicin derivative, and amanitin.

[0165] In some embodiments, the antibody or antigen-binding fragment thereof that has binding specificity for integrin alpha10 comprises a doxorubicin derivative that is 3'-deamino-3"-4'-anhydro-[2"(S)-methoxy-3"(R)-hydroxy-4"-morpholinyl]doxorubicin.

[0166] In some embodiments, the cytotoxic moiety is a chemotherapeutic agent. For example, in some embodiments, the chemotherapeutic agent can be an alkylating agent, an antimetabolite, an anti-microtubule inhibitor, a topoisomerase inhibitor, or a cytotoxic antibiotic. For example, in some embodiments, the chemotherapeutic agent can be selected from the group consisting of anthracyclines, taxanes, and platinum agents. For example, in some embodiments, the chemotherapeutic agent can be selected from the group consisting of cisplatin, paclitaxel, albumin-bound paclitaxel, docetaxel, cyclophosphamide, epirubicin, doxorubicin, carboplatin, gemcitabine, bleomycin, fluorouracil, cyclophosphamide, vinorelbine, capecitabine, ixabepilone, and ixabepilone, or combinations thereof.

[0167] In some embodiments, the transcriptional toxin is selected from the group consisting of shiga and shiga-like toxins, type I ribosome-inactivating proteins, type II ribosome-inactivating proteins, and saporin, or combinations thereof.

[0168] In some embodiments, the type I ribosome-inactivating protein is trichosanthin and / or raffin.

[0169] In some embodiments, the type II ribosome-inactivating protein is ricin, agglutinin, and / or abrin.

[0170] In some embodiments, the cytotoxic drug is suitable for use in activation therapy, such as photon activation therapy, neutron activation therapy, neutron induced Auger electron therapy, synchrotron radiation therapy, or low energy X-ray photon activation therapy.

[0171] In some embodiments, the antibody or antigen-binding fragment thereof that has binding specificity for integrin alphal0 comprises a detectable moiety.

[0172] In some embodiments, the detectable moiety is selected from the group consisting of a fluorophore, an enzyme, and a radioactive tracer or radioisotope.

[0173] In some embodiments, the detectable moiety comprises or consists of a radioisotope.

[0174] In some embodiments, the radioisotope is 99m Tc, 111 In, 67 Ga, 68Ga , 72 As, 89 Zr, 123 I, and 201 Tl.

[0175] In some embodiments, the antibody or antigen-binding fragment thereof that has binding specificity for integrin alpha 10 is 86 Y / 90 Y or 12 4I / 211 It contains pairs of detectable and cytotoxic radioisotopes such as At.

[0176] In some embodiments, an antibody or antigen-binding fragment thereof having binding specificity for integrin alpha10 comprises a radioisotope that can act simultaneously as a detectable moiety and as a cytotoxic moiety in a multimodal manner.

[0177] In some embodiments, the detectable moiety comprises or consists of a paramagnetic isotope.

[0178] In some embodiments, the paramagnetic isotope is 157 Gd, 55 Mn, 162 Dy, 52 Cr, and 56 Fe.

[0179] In some embodiments, the detectable moiety is detectable by imaging techniques such as SPECT, PET, MRI, light or ultrasound imaging.

[0180] In some embodiments, the cytotoxic moiety and / or the detectable moiety are indirectly attached to the antibody or antigen-binding fragment thereof via a linking moiety.

[0181] In some embodiments, the linking moiety is a chelator.

[0182] In some embodiments, the chelating agent is selected from the group consisting of derivatives of 1,4,7,10-tetraazacyclododecane-1,4,7,10,tetraacetic acid (DOTA), deferoxamine (DFO), derivatives of diethylenetriaminepentaacetic acid abid (DTPA), derivatives of S-2-(4-isothiocyanatobenzyl)-1,4,7-triazacyclononane-1,4,7-triacetic acid (NOTA), and derivatives of 1,4,8,11-tetraazacyclodocedane-1,4,8,11-tetraacetic acid (TETA).

[0183] In some embodiments, the antibody or antigen-binding fragment thereof that has binding specificity for integrin alphal0 does not comprise a cytotoxic or detectable moiety.

[0184] Antibody production Another aspect of the invention relates to a polynucleotide encoding the antibody or antigen-binding fragment of the first aspect of the invention, or its component polypeptide chains.

[0185] "Polynucleotide" includes DNA (eg, genomic or complementary DNA) and mRNA molecules, which may be single-stranded or double-stranded.

[0186] In some embodiments, the polynucleotide is an isolated polynucleotide.

[0187] In some embodiments, the polynucleotide is a cDNA molecule.

[0188] One of skill in the art will appreciate that a polynucleotide can be codon optimized for expression of an antibody or antigen-binding fragment thereof in a particular host cell, for example, for expression in a human cell (see, e.g., Angov, 2011, Biotechnol. J. 6(6):650-659, the disclosure of which is incorporated herein by reference).

[0189] In some embodiments, a polynucleotide encoding an antibody or antigen-binding fragment of the invention encodes an antibody light chain or a variable region thereof.

[0190] In some embodiments, a polynucleotide encoding an antibody or antigen-binding fragment of the invention encodes an antibody heavy chain or a variable region thereof.

[0191] Another aspect of the invention relates to a vector comprising a polynucleotide according to another aspect of the invention.

[0192] In some embodiments, the vector is an expression vector.

[0193] The term "expression vector" is defined as a DNA molecule, e.g., linear or circular, that contains a polynucleotide encoding a polypeptide of the invention (an antibody or antigen-binding fragment thereof) or is operably linked to additional nucleotides provided for its expression. The terms "plasmid," "expression vector," and "vector" are used interchangeably, as the plasmid is currently the most commonly used form of vector. However, the invention is intended to include other forms of expression vectors that serve equivalent functions.

[0194] Another aspect of the invention relates to a recombinant host cell comprising a polynucleotide according to another aspect of the invention or a vector according to another aspect of the invention.

[0195] In some embodiments, the recombinant host cell is a bacterial cell.

[0196] In some embodiments, the recombinant host cell is a yeast cell.

[0197] In some embodiments, the recombinant host cell is a mammalian cell.

[0198] In some embodiments, the recombinant host cell is a human cell.

[0199] In another aspect, the invention relates to a method for producing an antibody or antigen-binding fragment thereof according to another aspect of the invention, the method comprising culturing a host cell according to another aspect of the invention, a polynucleotide according to another aspect of the invention, or a vector of the third aspect of the invention under conditions permitting expression of the encoded antibody or antigen-binding fragment thereof.

[0200] Detection of integrin alpha 10 Another aspect of the invention relates to an in vivo method for the detection of cells expressing integrin alpha10 beta1 in a subject, the method comprising: (a) providing a sample of cells from the subject to be tested, such as a tissue biopsy or blood sample; (b) optionally extracting and / or purifying cells present in the sample; (c) contacting the antibody or antigen-binding fragment of the first aspect of the invention with cells present in the sample; (d) determining whether the antibody or antigen-binding fragment thereof binds to the cell; Binding of the antibody or antigen-binding fragment thereof to the cells indicates the presence of a disease or disorder associated with cells expressing integrin alpha10 beta1 in the tissue of the subject.

[0201] In some embodiments, the antibody is covalently linked to a detectable moiety, and the detectable moiety is selected from the group consisting of a fluorophore, an enzyme, and a radioactive tracer or a radioisotope. The integrin alpha 10 antigen can also be detected by detecting a peptide, protein, or polypeptide other than the integrin alpha 10 polypeptide, which can specifically bind to the integrin alpha 10 antigen. In some embodiments, the peptide, protein, or polypeptide is linked to an enzyme, a fluorophore, or a radioactive tracer. The radioactive tracer can be selected from, for example, a positron emitter or a gamma emitter. The conjugation of the antibody to a detectable moiety can facilitate and improve the detection of the antibody, which in turn can facilitate the detection of integrin alpha 10-expressing cells in a sample, and thus the diagnosis of cancer.

[0202] In some embodiments, the antibodies of the present disclosure can be used to detect integrin alpha10 in vitro, such as by in vivo antibody-based detection techniques described herein and / or known to those of skill in the art, or even in vivo and / or in situ, on cells, in tissues, in blood of samples obtained from a mammal.

[0203] Those skilled in the art can select standard laboratory setups for the detection of integrin alphal0 antibodies depending on the context and physical state of the sample.

[0204] In some embodiments, one of skill in the art would perform the detection step using flow cytometry, such as fluorescence activated cell sorting (FACS).

[0205] Exemplary immunological methods well known in the art include, but are not limited to, Western blot, enzyme-linked immunosorbent assay (ELISA), radioimmunoassay (RIA), immunohistochemistry (IHC), immunofluorescence assay (IF), and fluorescent in situ hybridization (FISH).

[0206] Detection of integrin alpha10 can be accomplished using methods well known in the art of detection and imaging, such as conventional fluorescence microscopy, confocal microscopy, two-photon microscopy, clinical imaging such as stimulated emission depletion (STED), and the like.

[0207] In some embodiments, the detectable moiety is selected from the group consisting of a fluorophore, an enzyme, or a radioactive tracer.

[0208] Typical methods for detecting cell surface antigens in vivo are well known in the art and include, but are not limited to, fluorescence imaging, positron emission tomography, X-ray computed tomography (CT), magnetic resonance imaging (MRI) and functional magnetic resonance imaging (fMRI), ultrasound, and single photon emission computed tomography (SPECT).In particular, cell surface antigens can be imaged in vivo using immunolabeling with a radioactive tracer bound to an antibody or other specific binding protein.

[0209] In some embodiments, antibodies used for in vivo imaging are antibody fragments, such as Fab fragments, and single chain antibodies, due to their small size and lack of effector functions.

[0210] Pharmaceutical Compositions and Administration Thereof Another aspect of the invention relates to a pharmaceutical composition comprising an effective amount of: An antibody or antigen-binding fragment according to another aspect of the present invention. A polynucleotide according to another aspect of the present invention, A vector according to another aspect of the invention, and / or A host cell according to another aspect of the present invention, In a pharmaceutical composition, the composition further comprises a pharma- ceutically acceptable buffer, diluent, carrier, or excipient.

[0211] In some embodiments, the present disclosure relates to a composition, such as a pharmaceutical composition, comprising: The composition comprises an antibody that specifically binds to an integrin alphal0 polypeptide or a fragment thereof, and is for use in the diagnosis and / or treatment of a form of cancer as defined herein.

[0212] In some embodiments, the present disclosure relates to a composition, such as a pharmaceutical composition, The composition is for use in the diagnosis and / or treatment of a cancer form selected from the group consisting of breast cancer, brain cancer, cancer of the CNS, lung cancer, prostate cancer, pancreatic cancer, skin cancer, lymphoma and sarcoma, or a metastasis of any one of said cancer forms. Detailed definitions of cancer forms that can be treated and / or diagnosed using the antibodies and compositions of the present disclosure are found elsewhere herein, for example in the "Clinical Conditions" section.

[0213] In some embodiments, compositions for use in the diagnosis and / or treatment of cancer forms defined herein comprise a pharma- ceutical effective amount, such as a pharmacologically effective concentration, of an antibody that specifically binds to an integrin alphal0 polypeptide or a fragment thereof.

[0214] The pharmacologically effective concentration referred to herein is typically the concentration of integrin alphal0 antibody that induces a desired response in an individual receiving the pharmaceutical composition.

[0215] In some embodiments, compositions for use in the diagnosis and / or treatment of cancer forms defined herein comprise a pharmacologically effective concentration of an antibody that specifically binds to an integrin alphal0 polypeptide or a fragment thereof, wherein the antibody or fragment thereof is conjugated to an additional moiety.

[0216] For example, the additional moiety can be a detectable moiety. An antibody that specifically binds to an integrin alpha 10 polypeptide or a fragment thereof and is conjugated to a detectable moiety can be useful for detecting the expression of integrin alpha 10 on a cell and thus determining whether said cell is a malignant cell and / or a tumor-associated cell.

[0217] For example, the additional moiety can be a cytotoxic moiety. Antibodies that specifically bind to an integrin alphal0 polypeptide or a fragment thereof and are conjugated to a cytotoxic moiety, for example, antibody drug conjugates (ADCs) comprising an antibody that specifically binds to an integrin alphal0 polypeptide or a fragment thereof, can be particularly useful for targeting a particular cytotoxic moiety and / or drug to cells that express integrin alphal0 and are malignant and / or tumor-associated cells.

[0218] ADCs can be generated using well-established conjugation techniques, for example, targeting antibodies can be conjugated to DNA intercalator and topoisomerase inhibitor type payloads by mild reduction of inter- and / or intra-chain disulfide bonds.

[0219] For example, the additional moiety can include a biological response modifier. A biological response modifier is a substance that modifies the immune response by enhancing or suppressing the immune response. The biological response modifier can be endogenous, such as a moiety normally produced naturally in the body, or can be exogenous.

[0220] In some embodiments, the additional moiety may include a biological response modifier, such as a cytokine, lymphokine, interferon, or a combination thereof.

[0221] The composition for use in the present disclosure may be a pharmaceutical composition suitable for parenteral administration. Such compositions preferably include aqueous and non-aqueous sterile injection solutions that may contain wetting or emulsifying agents, antioxidants, pH buffers, bacteriostatic compounds, and solutes that render the formulation isotonic with solid liquids, preferably blood, and aqueous and non-aqueous sterile suspensions that may contain suspending agents and thickening agents. The pharmaceutical compositions may be provided in unit-dose or multi-dose containers, such as sealed ampoules and vials, and may be stored in a freeze-dried state, requiring only the addition of a sterile liquid carrier immediately prior to use, for injection immediately prior to use.

[0222] In some embodiments, compositions for use of the present disclosure further comprise at least one of a pharma- ceutically acceptable buffer, diluent, carrier, or excipient.

[0223] Preferably, the composition of the present invention comprises one or more suitable pharmaceutical excipients for use with cells, tissues or organisms, such as pharmaceutical excipients suitable for administration to individuals, which may be non-sterile or sterile.Such excipients may include, but are not limited to, saline, buffered saline, dextrose, water, glycerol, ethanol, and combinations of these excipients in various amounts.The formulation should be suitable for the mode of administration.

[0224] Preferably, the pharmaceutical compositions of the present invention are prepared in an injectable form, either as liquid solutions or suspensions; solid forms suitable for dissolution or suspension in liquid prior to injection are also within the scope of the present invention. The preparation may also be emulsified or encapsulated in liposomes.

[0225] Integrin alphal0 polypeptide antibodies, or polynucleotides that specifically bind to a polynucleotide transcript encoding an integrin alphal0 polypeptide, may be administered alone or in combination with other compounds, in any order, either simultaneously or sequentially.

[0226] Administration can be parenteral, for example, via injection or infusion. Parenteral injection can be, for example, intraventricular, intrathecal, intratumoral, intravenous, intramuscular, intradermal, or subcutaneous injection. Preferably, the administration is parenteral, via injection or infusion.

[0227] Those skilled in the art will appreciate that additional compounds, including chelating agents such as EDTA, citrate, EGTA, or glutathione, may also be included in the pharmaceutical composition.

[0228] Pharmaceutical compositions can be prepared by methods well known in the art that are sufficiently storage stable and suitable for administration to humans and animals, for example, pharmaceutical compositions can be lyophilized, for example, by freeze drying, spray drying, spray cooling, or by using particle formation from supercritical particle formation.

[0229] The excipient may be one or more of carbohydrates, polymers, lipids, and minerals. Examples of carbohydrates include lactose, glucose, sucrose, mannitol, and cyclodextrin, which are added to the composition, for example, to facilitate lyophilization. Examples of polymers are starch, cellulose ether, cellulose carboxymethylcellulose, hydroxypropylmethylcellulose, hydroxyethylcellulose, ethylhydroxyethylcellulose, alginates, carrageenans, hyaluronic acid and its derivatives, polyacrylic acid, polysulfonates, polyethylene glycol / polyethylene oxide, polyethylene oxide / polypropylene oxide copolymers, polyvinyl alcohol / polyvinyl acetate with different hydrolysis degrees, and polyvinylpyrrolidone, all with different molecular weights, which are added to the composition, for example, to adjust viscosity, bioadhesion, or to protect lipids from chemical and proteolytic degradation. Examples of lipids are fatty acids, phospholipids, mono-, di- and triglycerides, ceramides, sphingolipids and glycolipids of all different acyl chain lengths and degrees of saturation, egg lecithin, soy lecithin, hydrogenated egg and soy lecithin, which are added to the composition for reasons similar to those of the polymers. Examples of minerals are talc, magnesium oxide, zinc oxide, and titanium oxide, which are added to the composition to obtain benefits such as reduced liquid accumulation or favorable pigment properties.

[0230] The antibodies or antigen-binding fragments of the present invention can be formulated into any type of pharmaceutical composition known in the art suitable for their administration.

[0231] In some embodiments, the pharmaceutical composition of the present invention may be in the form of liposomes, in which the antibody or antigen-binding fragment is combined with an amphiphilic agent, such as lipids, which exist in aggregated form as micelles, insoluble monolayers, and liquid crystals, in addition to other pharma- ceutically acceptable carriers. Suitable lipids for liposomal formulations include, but are not limited to, monoglycerides, diglycerides, sulfatides, lysolecithin, phospholipids, saponins, bile acids, and the like. Suitable lipids also include the above lipids modified with poly(ethylene glycol) at the polar head group to extend bloodstream circulation time. Preparation of such liposomal formulations can be found, for example, in US 4,235,871, the disclosure of which is incorporated herein by reference.

[0232] The pharmaceutical composition of the present invention can also be in the form of biodegradable microspheres. Aliphatic polyesters such as poly(lactic acid) (PLA), poly(glycolic acid) (PGA), copolymers of PLA and PGA (PLGA), or poly(caprolactone) (PCL), and polyanhydrides are widely used as biodegradable polymers for the manufacture of microspheres. Preparations of such microspheres can be found in US5,851,451 and EP0213303, the disclosures of which are incorporated herein by reference.

[0233] In some embodiments, the pharmaceutical composition of the present invention is provided in the form of a polymer gel, and polymers such as starch, cellulose ether, cellulose carboxymethylcellulose, hydroxypropylmethylcellulose, hydroxyethylcellulose, ethylhydroxyethylcellulose, alginates, carrageenans, hyaluronic acid and its derivatives, polyacrylic acid, polysulfonates, polyethylene glycol / polyethylene oxide, polyethylene oxide / polypropylene oxide copolymers, polyvinyl alcohol / polyvinyl acetate with different hydrolysis degrees, and polyvinylpyrrolidone are used to give viscosity to the solution containing the formulation.These polymers may also include gelatin or collagen.

[0234] Alternatively, the antibody or antigen-binding fragment may be simply dissolved in saline, water, polyethylene glycol, propylene glycol, ethanol or oils (such as safflower oil, corn oil, peanut oil, cottonseed oil, or sesame oil), tragacanth gum, and / or various buffers.

[0235] It will be understood that the pharmaceutical compositions of the present invention may contain ions and a defined pH to enhance the action of the active antibody or antigen-binding fragment. Furthermore, the compositions may be subjected to conventional pharmaceutical operations such as sterilization and / or may contain conventional auxiliary agents such as preservatives, stabilizers, wetting agents, emulsifiers, buffers, fillers, etc.

[0236] The pharmaceutical compositions according to the invention can be administered by any suitable route known to those skilled in the art. Possible routes of administration thus include parenteral (intravenous, subcutaneous and intramuscular), topical, intraocular, nasal, pulmonary, buccal, buccal, parenteral, vaginal and rectal. Administration via implants is also possible.

[0237] In a preferred embodiment, pharmaceutical compositions can be administered parenterally, for example, intravenously, intraventricularly, intraarticularly, intraperitoneally, intrathecally, intraventricularly, intrasternally, intracranially, intramuscularly, or subcutaneously, or by infusion techniques.They are conveniently used in the form of a sterile aqueous solution, which may contain other substances, for example, sufficient salts or glucose to make the solution isotonic with blood.The aqueous solution should be appropriately buffered as necessary.The preparation of suitable parenteral formulations under sterile conditions can be easily achieved using standard pharmaceutical techniques well known to those skilled in the art.

[0238] Preparations suitable for parenteral administration include aqueous and non-aqueous sterile injection solutions that may contain antioxidants, buffers, bacteriostats, and solutes that render the preparation isotonic with the blood of the intended recipient, as well as aqueous and non-aqueous sterile suspensions that may contain suspending agents and thickening agents.The preparations may be presented in unit-dose or multi-dose containers, such as sealed ampoules and vials, and may be stored in a freeze-dried (lyophilized) condition, requiring only the addition of a sterile liquid carrier, such as water, for injection immediately prior to use.Extemporaneous injection solutions and suspensions may be prepared from sterile powders, granules, and tablets of the kind previously described.

[0239] Thus, the pharmaceutical compositions of the present invention are particularly suitable for parenteral, eg, intravenous, administration.

[0240] The pharmaceutical compositions are administered to a patient in pharmacologically effective doses.

[0241] As used herein in the context of diagnostic uses of the antibodies or antigen-binding fragments of the invention, a "pharmacologically effective amount" or a "pharmacologically effective concentration" or an "effective concentration" or an "effective amount" or a "diagnostically effective" refers to an amount that provides a detectable signal for diagnosis (e.g., for in vivo imaging purposes).

[0242] Those skilled in the art will appreciate that the pharmaceutical compositions of the present invention may be administered alone or in combination with other therapeutic agents used in the treatment of neoplastic disorders or diseases.

[0243] In some embodiments, the composition is adapted for parenteral delivery.

[0244] In some embodiments, the composition is adapted for intravenous delivery.

[0245] In some embodiments, the composition is adapted for local delivery.

[0246] In some embodiments, the composition is adapted for subcutaneous delivery.

[0247] In some embodiments, the composition is adapted for intramuscular delivery.

[0248] Those skilled in the art will further appreciate that the antibodies or antigen-binding fragments and pharmaceutical compositions or formulations of the invention have utility in both the medical and veterinary fields. Thus, the methods of the invention can be used to treat both humans and non-human animals, such as horses, dogs, and cats. Preferably, however, the patient is a human.

[0249] Indications and uses of antibodies Another aspect of the present invention is a method for producing a An antibody or antigen-binding fragment according to another aspect of the present invention. A polynucleotide according to another aspect of the present invention, A vector according to another aspect of the present invention. A host cell according to another aspect of the invention, and / or The composition according to another aspect of the present invention, Regarding use in medicine.

[0250] Another aspect of the present invention is a method for producing a An antibody or antigen-binding fragment according to another aspect of the present invention. A polynucleotide according to another aspect of the present invention, A vector according to another aspect of the present invention. A host cell according to another aspect of the invention, and / or The composition according to another aspect of the present invention, For use in the prevention and / or treatment and / or alleviation and / or detection and / or diagnosis of a disease or disorder susceptible to treatment with an inhibitor of integrin alpha10 beta1, which disease or disorder is associated with cells expressing integrin alpha10 beta1.

[0251] Depending on whether it is therapeutically desirable to kill target cells expressing integrin alpha10 beta1, for example in the case of tumor cells, an antibody or antigen-binding fragment according to the first aspect of the invention capable of inducing ADCC and / or ADCP and / or CDC may be used. For example, if the target cells expressing integrin alpha10 beta1 are cancer cells, it may be advantageous for the antibody or antigen-binding fragment to be capable of inducing ADCC in order to eliminate such cells.

[0252] In one embodiment, the antibodies or antigen-binding fragments of the disclosure activate an ADCC response.

[0253] However, it will be understood that therapeutic benefit may still be achieved using antibodies or antigen-binding fragments that lack ADCC activity.

[0254] In one aspect of the invention, the present invention comprises: An antibody or antigen-binding fragment according to another aspect of the present invention. A polynucleotide according to another aspect of the present invention, A vector according to another aspect of the present invention. A host cell according to another aspect of the invention, and / or The composition according to another aspect of the present invention, The present invention relates to use in the prevention and / or treatment and / or alleviation and / or detection and / or diagnosis of diseases or disorders which are susceptible to treatment with inhibitors against integrin alpha10 beta1.

[0255] In an aspect of the invention, the present invention provides an antibody or antigen-binding fragment according to another aspect of the invention, A polynucleotide according to another aspect of the present invention, A vector according to another aspect of the present invention. A host cell according to another aspect of the invention, and / or The composition according to another aspect of the present invention, For use in the prevention and / or treatment and / or alleviation and / or detection and / or diagnosis of a disease or disorder, said disease or disorder being associated with cells expressing integrin alpha10 beta1.

[0256] In some embodiments, the disease or disorder is associated with integrin alpha10beta1 signaling.

[0257] With respect to the therapeutic and prophylactic aspects of the invention, those skilled in the art will appreciate that binding of an antibody or antigen-binding fragment thereof to integrin alpha 10 present on the surface of cells associated with a neoplastic disease or disorder may result in modulation (i.e., increase or decrease) of the biological activity of integrin alpha 10 beta 1. However, such a modulating effect is not required, for example, an antibody or antigen-binding fragment thereof of the invention may induce a therapeutic and prophylactic effect simply by binding to integrin alpha 10 on the surface of cells associated with a disease or disorder, which may in turn trigger the immune system to induce processes such as cell death (e.g., by ADCC and / or by the presence within the agent of a cytotoxic / radioactive moiety).

[0258] In some embodiments the antibody or antigen-binding fragment thereof that has binding specificity for integrin alpha10 inhibits the biological activity of integrin alpha10 beta1.

[0259] "Integrin alphal0 beta1 biological activity" includes any interaction or signaling event involving integrin alphal0 beta1 on cells associated with a neoplastic disease or disorder. Some biological activities associated with cells associated with a neoplastic disease or disorder that may be inhibited by the antibodies, antigens, or fragments thereof of the present disclosure include cell adhesion, cell proliferation, cell migration, cell survival, tumor growth and / or metastasis, integrin alphal0 beta1 signaling, and / or any combination thereof. These are described herein in the "Inhibition of Biological Activity" section.

[0260] Another aspect of the invention is an antibody or antigen-binding fragment according to another aspect of the invention, A polynucleotide according to another aspect of the present invention, A vector according to another aspect of the present invention. A host cell according to another aspect of the invention, and / or The composition according to another aspect of the present invention, For use in inhibiting cell survival, cell migration, cell proliferation, cell growth and / or cell adhesion, tumor growth, and metastasis, the target cells express integrin alpha10 beta1.

[0261] Another aspect of the invention relates to a method for in vivo imaging of integrin alpha10 beta1 expression in a mammal, the method comprising: a) providing a mammal; b) providing an antibody or antigen-binding fragment thereof disclosed herein; c) administering the antibody or antigen-binding fragment thereof to a mammal such that the antibody or fragment thereof is capable of binding to the extracellular domain of integrin alpha10 beta1 of cells in the mammal; d) optionally adding a second, labeled antibody or fragment thereof to the sample, said second antibody or fragment thereof binding to the antibody or fragment thereof of c); e) detecting the antibody or antigen-binding fragment thereof on said cells in c) or, optionally, detecting a second, labeled antibody or fragment thereof in d) bound to the antibody or fragment thereof; generating an image of the detected antibody or fragment thereof, thereby imaging the expression of integrin alpha10 beta1 on mammalian cells in vivo.

[0262] In vitro diagnostic assays based on the antibodies or antigen-binding fragments thereof disclosed herein can be used to detect the presence of integrin alpha 10 or fragments thereof on the surface of extracellular vesicles such as exosomes that have left a tumor site, and such assays can also be used to detect the presence of integrin alpha 10 or fragments thereof that are no longer bound to cells or vesicles, for example, in the case of shedding of cells such as tumor cells. For example, the antibodies or antigen-binding fragments thereof disclosed herein can be used to detect the presence of integrin alpha 10 on the surface of extracellular vesicles such as exosomes, as well as fragments of integrin alpha 10 found in blood and / or plasma.

[0263] Therefore, another aspect of the present invention relates to an in vitro method for detecting integrin alphal0 or a fragment thereof in a sample obtained from a mammal, said method comprising the steps of: a) providing a sample obtained from a mammal; b) providing an antibody or antigen-binding fragment thereof disclosed herein; c) contacting the antibody or antigen-binding fragment thereof with the sample such that the antibody or fragment thereof is capable of binding to the extracellular domain of integrin alphal0, or a fragment thereof, in the sample; d) optionally adding a second, labeled antibody or fragment thereof to the sample, said second antibody or fragment thereof binding to the antibody or fragment thereof of c); e) detecting the antibody or antigen-binding fragment thereof in c) or, optionally, detecting a second, labeled antibody or fragment thereof in d) bound to the antibody or fragment thereof; Thereby, it is determined whether integrin alpha 10 or a fragment thereof is found in a sample obtained from the mammal.

[0264] For example, the sample can be a mammalian bodily fluid, such as blood, plasma, cerebrospinal fluid, urine, etc.

[0265] In some embodiments, the sample comprises cells, extracellular vesicles and / or fragments thereof.

[0266] Clinical status In some embodiments, the cancer type being treated and / or prevented and / or detected and / or diagnosed and / or classified and / or prognosed and / or metastasis is prevented is selected from the group consisting of breast cancer, brain cancer, cancer of the central nervous system (CNS), lung cancer, prostate cancer, pancreatic cancer, skin cancer, lymphoma, and sarcoma.

[0267] In some embodiments, the breast cancer is a triple-negative breast cancer morphology, the triple-negative breast cancer being selected from the group consisting of Basal-like 1 breast cancer, Basal-like 2 breast cancer, claudin-low breast cancer, metastatic breast cancer (MBC), interferon-rich breast cancer, immunomodulatory breast cancer, mesenchymal breast cancer, mesenchymal stem-like breast cancer, luminal androgen receptor breast cancer, and unstable breast cancer.

[0268] In some embodiments, the breast cancer is a triple-negative breast cancer morphology and has morphological characteristics of invasive ductal carcinoma.

[0269] In some embodiments, the breast cancer is a triple-negative breast cancer morphology and has morphological characteristics of Basal-like triple-negative breast cancer.

[0270] In some embodiments, the breast cancer is a basal-like breast cancer morphology, having morphological characteristics of invasive ductal carcinoma.

[0271] In some embodiments, the cancer is prostate cancer, wherein the prostate cancer is small cell (neuroendocrine) carcinoma (SCNC) or castration-resistant prostate cancer (CRPC).

[0272] In some embodiments, the cancer is lung cancer, wherein the lung cancer is squamous cell carcinoma, lung adenocarcinoma, small cell lung cancer, or large cell lung cancer.

[0273] In some embodiments, the cancer is pancreatic cancer, which is an exocrine or endocrine tumor.

[0274] In some embodiments, the pancreatic cancer is an exocrine tumor selected from the group consisting of ductal adenocarcinoma, acinar cell carcinoma, adenosquamous carcinoma, intraductal papillary mucinous neoplasm (IPMN), and pancreatic intraepithelial neoplasia.

[0275] In some embodiments, the pancreatic cancer is selected from the group consisting of neuroendocrine tumors, gastrinomas, glucagonomas, insulinomas, somatostatinomas, VIP adenomas, and nonfunctioning islet cell tumors. In some embodiments, the cancer is pancreatic cancer, and the pancreatic cancer is a neuroendocrine tumor.

[0276] In some embodiments, the cancer is pancreatic cancer, and the pancreatic cancer is grade I, grade II, or grade III pancreatic cancer.

[0277] In some embodiments, the sarcoma is selected from the group consisting of chondrosarcoma, osteosarcoma, undifferentiated pleomorphic sarcoma, myxosarcoma, dedifferentiated liposarcoma, atypical lipomatous tumor, myxoinflammatory fibroblastoma, low-grade fibromyosarcoma, sclerosing epithelial fibrosarcoma, pseudomyogenic chondrosarcoma, and sclerosing myeloid tumor.

[0278] In some embodiments, the brain cancer and / or CNS cancer is: a) Neuroepithelial tumors selected from the following: i) Astrocytic tumors selected from the following: Pilocytic astrocytoma, pyromyxoid astrocytoma, pituitary giant cell astrocytoma, pleomorphic xanthoastrocytoma, diffuse astrocytoma, anaplastic astrocytoma, glioblastoma, giant cell glioblastoma, gliosarcoma, cerebral gliomatosis, ii) oligodendroglial tumors selected from oligodendroglioma and anaplastic oligodendroglioma; and iii) oligodendroglioma and anaplastic oligodendroglioma; iv) ependymoma, myxopapillary ependymoma, ependymoma, anaplastic ependymoma, and v) choroid plexus tumors selected from choroid plexus papillomas; Atypical choroid plexus papillomas and choroid plexus tumors vi) other neuroepithelial tumors selected from astroblastoma, choroid plexus glioma of the third ventricle, and angiocentric glioma; vii) Neuronal and mixed neuroglial tumors selected from cerebellar glioma (Lhermitte-Duclos), lytic childhood astrocytoma / glioma, pigmented neuroepithelial tumor, glioma, glioma, anaplastic glioma, central neurocytoma, extraventricular neurocytoma, cerebellar lipocytoma; Papillary glioma, rosette-forming glioma of the fourth ventricle, and paraganglioma, viii) Tumors of the pineal region selected from pineocytoma, moderately differentiated pineal parenchymal tumor, pineoblastoma, and papillary tumor of the pineal region; ix) Embryonal tumors selected from medulloblastoma, medulloblastoma with widespread nodularity, anaplastic medulloblastoma, CNS primitive neuroectodermal tumor, CNS neuroblastoma, and atypical teratoid / rhabdoid tumor; b) Tumors of the cranial nerves and paraspinal nerves selected from the following: i) schwannoma, ii) neurofibroma, iii) perineurioma, and iv) malignant peripheral nerve sheath tumor (MPNST), c) Meningeal tumors selected from the following: i) a tumor of meningeal cells selected from meningioma, atypical meningioma, and anaplastic meningioma; ii) lipoma, angiolipoma, hibernoma, liposarcoma, solitary fibrous tumor, fibrosarcoma, malignant fibrous histiocytoma, leiomyoma, leiomyosarcoma, rhabdomyoma, rhabdomyosarcoma, chondroma, chondrosarcoma, osteoma, osteosarcoma, osteochondroma, plasma, epithelioid hemangioendothelioma, plasmathelioma, undifferentiated pyloric epithelioma, and mesenchymal tumors selected from angiosarcoma, Kaposi's sarcoma, E-wing sarcoma-PNET; iii) primary melanocytic lesions selected from diffuse melanocytosis, melanocytoma, malignant melanoma, meningeal melanomatosis; iv) other neoplasms involving the meninges such as hemangioblastoma; d) Hematopoietic tumors selected from the following: i) malignant lymphoma, phytosarcoma, and ii) granulocytic sarcoma, e) Tumors in the outcrop area selected from the following: i) craniopharyngioma, ii) granular cell tumor, iii) pituitary cell tumor, and iv) adenohypophysial spindle cell tumor.

[0279] The above list of CNS tumors is based on the more detailed, more complete and more up-to-date CNS tumor classification found in Louis et al. (2021) The WHO Classification of Tumours of the Nervous System: a summary. Neuro-Oncology, Volume 23, Issue 8, August 2021, Pages 1231-1251.

[0280] In some embodiments, the brain cancer and / or CNS cancer is a glioma.

[0281] In some embodiments, the brain cancer and / or CNS cancer is grade II, III, or IV glioma.

[0282] In some embodiments, the brain cancer and / or CNS cancer is astrocytoma, for example, grade II astrocytoma, grade III astrocytoma, or grade IV astrocytoma.

[0283] In some embodiments, the glioma is a glioblastoma.

[0284] In some embodiments, the glioma is a primary glioblastoma.

[0285] In some embodiments, the glioma is a secondary glioblastoma.

[0286] In some embodiments, the brain cancer and / or CNS cancer is medulloblastoma.

[0287] In some embodiments, the brain cancer and / or CNS cancer is neuroblastoma.

[0288] In some embodiments, the brain cancer and / or CNS cancer is selected from the group consisting of astrocytoma, anaplastic astrocytoma, cerebral hemangiocytoma, meningioma, hemangiomatous hemangioma, atypical meningioma, fibroblastic meningioma, meningoepithelioma, secretory meningioma, oligoastrocytoma, anaplastic oligoastrocytoma, oligodendroglioma, and anaplastic oligodendroglioma.

[0289] In some embodiments, the brain cancer and / or CNS cancer is selected from the group consisting of astrocytic tumors, oligodendroglial tumors, ependymal cell tumors, mixed gliomas, neuroepithelial tumors of unknown etiology, tumors of the choroid plexus, mixed neuronal and neuronal-glial tumors, pineal parenchymal tumors and tumors containing neuroblastic or glioblastoma elements (germinomas), ependymomas, astrocytomas, oligodendroglioma, oligoastrocytomas, neuroepithelial tumors, and mixed neuronal and neuronal-glial tumors.

[0290] In some embodiments, the cancer is a metastasis, for example, a metastasis of any one of breast cancer, brain cancer, cancer of the CNS, lung cancer, prostate cancer, pancreatic cancer, skin cancer, lymphoma, and sarcoma.

[0291] In some embodiments, the cancer is a metastasis of any one of triple-negative breast cancer and inflammatory breast cancer.

[0292] In some embodiments, the cancer is a metastasis of a triple-negative breast cancer selected from the group consisting of Basal-like 1 breast cancer, Basal-like 2 breast cancer, claudin-low breast cancer, metastatic breast cancer (MBC), interferon-rich breast cancer, immunomodulatory breast cancer, mesenchymal breast cancer, mesenchymal stem-like breast cancer, luminal androgen receptor breast cancer, and unstable breast cancer.

[0293] In some embodiments, the cancer is a metastasis of lung cancer. In some embodiments, the cancer form is a metastasis of lung squamous cell carcinoma, lung adenocarcinoma, small cell lung carcinoma, or large cell lung carcinoma.

[0294] In some embodiments, the cancer is a metastasis of pancreatic cancer.

[0295] In some embodiments, the cancer is a metastasis of prostate cancer.

[0296] In some embodiments, the cancer is a metastasis of a sarcoma.

[0297] In some embodiments, the cancer is a brain cancer metastasis and / or a CNS cancer metastasis.

[0298] In some embodiments, the cancer is a brain cancer and / or a metastasis of a CNS cancer selected from the group consisting of astrocytoma, anaplastic astrocytoma, cerebral hemangiocytoma, meningioma, hemangiomatous hemangioma, atypical meningioma, fibroblastic meningioma, meningoepithelioma, secretory meningioma, oligoastrocytoma, anaplastic oligoastrocytoma, oligodendroglioma, and anaplastic oligodendroglioma.

[0299] Cancer Treatment Those skilled in the art will recognize that the antibodies or antigen-binding fragments thereof having binding specificity for integrin alpha10 disclosed by some embodiments of the present invention can be used to treat cancer.

[0300] In some embodiments, the antibody or antigen-binding fragment thereof has binding specificity for integrin alpha10 and thus binds to malignant cells and / or tumor-associated cells, e.g., cancer-associated fibroblasts (CAFs), stromal cells, stem cells and / or stem-like cells, and / or cells expressing integrin alpha10 beta1, e.g., tumor-associated macrophages (TAMs), immune cells, endothelial cells.

[0301] In some embodiments, the antibody or antigen-binding fragment thereof with binding specificity for integrin alpha10 is for use in the treatment of cancer, wherein treatment is initiated upon detection of integrin alpha10 polypeptide in cancer cells within a tumor of a subject.

[0302] In some embodiments, an antibody or antigen-binding fragment thereof having binding specificity for integrin alpha10 is administered to an individual in need thereof in combination with radiation therapy and / or surgical removal of the cancer, such as prior to radiation therapy and / or surgical removal, or after radiation therapy and / or surgical removal of the cancer.

[0303] Another aspect of the invention relates to an antibody or antigen-binding fragment of the first aspect of the invention, or a stem or progenitor cell thereof, for use in inducing cell death and / or inhibiting the growth and / or proliferation of pathological cells associated with a neoplastic disease in a subject, which cells express integrin alpha10 beta1.

[0304] Another aspect of the present invention is a method for producing a An antibody or antigen-binding fragment according to another aspect of the present invention. A polynucleotide according to another aspect of the present invention, A vector according to another aspect of the present invention. A host cell according to another aspect of the invention, and / or The composition according to another aspect of the present invention, For use in the preparation of a medicament for the prevention, treatment, mitigation, detection and / or diagnosis of a disease or disorder susceptible to treatment with an inhibitor of integrin alpha10 signaling, which disease or disorder is associated with cells expressing integrin alpha10 beta1.

[0305] Similarly, another aspect of the invention relates to a method for the prevention and / or treatment and / or alleviation and / or detection and / or diagnosis of a disease or disorder susceptible to treatment with an inhibitor of integrin alpha10 beta1 signaling, said disease or disorder being associated with cells expressing integrin alpha10 in a subject, said method comprising administering to a subject an effective amount of An antibody or antigen-binding fragment according to another aspect of the present invention. A polynucleotide according to another aspect of the present invention, A vector according to another aspect of the present invention. A host cell according to another aspect of the invention, and / or The method includes administering to a subject a composition according to another aspect of the invention.

[0306] Another aspect of the present invention is a method for producing a an antibody that specifically binds to an integrin alpha 10 polypeptide or a fragment thereof; or A composition comprising a polynucleotide that specifically binds to a polynucleotide transcript encoding an integrin alpha 10 polypeptide or a fragment or variant thereof, For use in the manufacture of a medicament for the treatment of cancer, the cancer is a cancer that expresses integrin alpha 10, for example the cancer is selected from the group consisting of breast cancer, brain cancer, cancer of the CNS, lung cancer, prostate cancer, pancreatic cancer, skin cancer, lymphoma, and sarcoma, or the cancer is metastatic.

[0307] Another aspect of the present disclosure relates to a method of treating cancer, wherein the cancer is selected from the group consisting of breast cancer, brain cancer, cancer of the CNS, lung cancer, prostate cancer, pancreatic cancer, skin cancer, lymphoma, and sarcoma, or the cancer is metastatic, the method comprising administering to a subject in need thereof a pharmacologic effective amount of a composition comprising: an antibody that specifically binds to an integrin alpha 10 polypeptide or a fragment thereof; or A polynucleotide that specifically binds to a polynucleotide transcript encoding an integrin alphal0 polypeptide or a fragment or variant thereof.

[0308] Another aspect of the invention relates to a method for treating a patient having a disease or disorder associated with cells expressing integrin alpha10 beta1, the method comprising: a) selecting a patient identified as having a disease or disorder associated with cells expressing integrin alpha10 beta1 using a method according to another aspect of the invention; b) administering to the patient a therapeutic agent effective to treat the disease or disorder.

[0309] Another aspect of the disclosure relates to a method of inhibiting integrin alpha10 beta1 mediated signaling of at least one cancer cell, the method comprising treating the at least one cancer cell with an effective amount of: a) an antibody or antigen-binding fragment specific for an integrin alpha 10 polypeptide, and / or b) contacting the subject with a composition comprising a polynucleotide transcript encoding an integrin alpha 10 polypeptide or a fragment or variant thereof; The at least one cancer cell is selected from the group consisting of breast cancer cells, brain cancer cells, cancer cells of a CNS cancer, lung cancer cells, prostate cancer cells, pancreatic cancer cells, skin cancer cells, lymphoma cells, sarcoma cells, and metastatic tumor cells, or any of the cancer types listed herein under "clinical conditions."

[0310] Those skilled in the art will appreciate that cell signaling includes the molecular mechanisms by which cells detect and respond to external stimuli. Cell signaling also includes transcriptional and translational control and mechanisms, as well as signal transduction mechanisms.

[0311] One aspect of the disclosure relates to a method of inhibiting a cellular function of at least one cancer cell, the method comprising contacting at least one cancer cell with an effective amount of a composition comprising: an antigen comprising an integrin alpha 10 polypeptide or a fragment thereof, and / or a polynucleotide transcript encoding an integrin alpha 10 polypeptide or a fragment or variant thereof, The at least one cancer cell is selected from the group consisting of breast cancer cells, brain cancer cells, cancer cells of a CNS cancer, lung cancer cells, prostate cancer cells, pancreatic cancer cells, skin cancer cells, lymphoma cells, sarcoma cells, and metastatic tumor cells, or any of the cancer types listed herein under "clinical conditions."

[0312] In some embodiments, the present invention provides a method for inhibiting the growth and / or proliferation of cells expressing integrin alpha10 beta1, comprising administering to a subject an effective amount of: an antibody that specifically binds to an integrin alpha 10 polypeptide or a fragment thereof; or The present invention includes polynucleotides that specifically bind to polynucleotide transcripts encoding integrin alphal0 polypeptides or fragments or variants thereof.

[0313] In some embodiments of the present disclosure, inhibiting at least one cancer cell comprises: inhibiting the proliferation of at least one cancer cell; inhibiting the growth of at least one cancer cell; Inhibiting self-renewal of at least one cancer cell; inhibiting anchorage-independent growth of at least one cancer cell; inhibiting the proliferation of at least one cancer cell; Inhibiting the migration of at least one cancer cell; inhibiting the survival of at least one cancer cell; Inhibiting adhesion of at least one cancer cell; and / or combinations thereof.

[0314] In another embodiment, inhibiting at least one cancer cell with i) an antibody that specifically binds to an integrin alpha 10 polypeptide, or ii) a polynucleotide that specifically binds to a polynucleotide transcript encoding an integrin alpha 10 polypeptide or a fragment or variant thereof inhibits anchorage-independent growth of at least one cancer cell.

[0315] In some embodiments, the present disclosure relates to a method of inhibiting at least one cancer cell, wherein the cancer cell is a metastatic tumor, the inhibiting comprising: metastatic tumor survival, Metastatic tumor growth, Metastatic tumor growth, Metastatic tumor migration, Metastatic tumor invasion, The onset of new metastatic tumors, Penetration of new metastatic tumors; and It's one of those combinations.

[0316] The cell may express one or more additional markers as defined herein. In some embodiments, the cell is a cancer-associated fibroblast (CAF), a stromal cell, a stem cell and / or a stem-like cell. The method can be performed in vitro or in vivo.

[0317] In some embodiments, the composition comprises an effective amount of: an antibody that specifically binds to an integrin alpha 10 polypeptide or a fragment thereof; or Polynucleotides that specifically bind to a polynucleotide transcript encoding an integrin alpha 10 polypeptide or a fragment or variant thereof, and that are capable of inducing cell death and / or inhibiting proliferation and / or inhibiting proliferation and / or inhibiting migration of cells expressing integrin alpha 10, are included.

[0318] In some embodiments, the treatment is initiated upon detection of integrin alphal0 polypeptide and / or polynucleotide transcripts in cancer cells within the subject's tumor.

[0319] In some embodiments, the methods disclosed herein target antigens comprising an integrin alphal0 polypeptide or a fragment thereof expressed on the surface of a cell.

[0320] One aspect of the present disclosure relates to a method for preventing metastasis from a primary cancer form selected from the group consisting of breast cancer, brain cancer, cancer of the CNS, lung cancer, prostate cancer, pancreatic cancer, skin cancer, lymphoma, and sarcoma, or a cancer type listed herein under "clinical conditions," comprising administering to a subject in need thereof a therapeutically effective amount of: a) an antibody or antigen-binding fragment specific for an integrin alpha 10 polypeptide, and / or b) administering a polynucleotide transcript encoding an integrin alphal0 polypeptide or a fragment or variant thereof.

[0321] One of skill in the art will appreciate that methods for preventing metastasis from a primary cancer can be administered upon detection of the primary cancer.

[0322] Detection and diagnosis of cancer morphology One aspect of the present disclosure relates to an agent comprising or consisting of an antibody having specificity against an integrin alpha 10 polypeptide or a fragment thereof for use in detecting cells associated with a mammalian cancer form, said cells expressing the integrin alpha 10 polypeptide, said cancer form being selected from the group consisting of breast cancer, brain cancer, cancer of the CNS, lung cancer, prostate cancer, pancreatic cancer, skin cancer, lymphoma, and sarcoma, as well as metastases of any one of said cancer forms.

[0323] Another aspect of the invention relates to an in vitro method for identifying a patient having a disease or disorder associated with cells expressing integrin alpha10 beta1 that would benefit from treatment with an antibody or antigen-binding fragment thereof of the first aspect herein, the method comprising: a) providing a sample, such as a tissue biopsy or blood sample, from a patient to be tested; b) optionally extracting and / or purifying cells present in the sample; c) contacting a cell with an antibody or antigen-binding fragment thereof as defined herein; d) determining whether the antibody or antigen-binding fragment thereof binds to the integrin alpha 10 domain or a fragment thereof; Binding of the antibody or antigen-binding fragment thereof to the integrin alphal0 domain or a fragment thereof is indicative of a patient who will benefit from treatment with an antibody or antigen-binding fragment thereof as defined herein.

[0324] Indeed, cells expressing integrin alpha10 beta1, such as cancer cells expressing integrin rapha10 beta1, may undergo shedding and / or degradation, for example as part of tumor tissue necrosis or its life cycle, which results in the cells and portions of the proteins expressed on their surface eventually reaching the bloodstream. Thus, cancer cells or cancer-associated cells expressing integrin alpha10 may be degraded and / or eliminated, and integrin alpha10 or fragments thereof may eventually find their way into the blood and be detected by the methods of the present disclosure by binding to the antibodies of the present disclosure.

[0325] Another aspect of the invention relates to an in vivo method for identifying a patient having a disease or disorder associated with cells expressing integrin alpha10 beta1 that would benefit from treatment with an antibody or antigen-binding fragment thereof of the first aspect herein, the method comprising: (a) contacting the antibody or antigen-binding fragment of the first aspect of the invention with cells present in a sample; (b) determining whether the antibody or antigen-binding fragment thereof binds to the cell; Binding of the antibody or antigen-binding fragment thereof to cells expressing integrin alpha10 beta1 is indicative of a patient who would benefit from treatment with an antibody or antigen-binding fragment according to another aspect of the invention.

[0326] In some embodiments, the present disclosure relates to a composition according to an aspect of the present disclosure for use in diagnosing a cancer form selected from the group consisting of breast cancer, brain cancer, cancer of the CNS, lung cancer, prostate cancer, pancreatic cancer, skin cancer, lymphoma and sarcoma, or a metastasis of any one of said cancer forms, which can be detected and / or diagnosed by using a composition according to an aspect of the present disclosure and are described in detail in the "Clinical Conditions" section.

[0327] Those skilled in the art will understand that the process of detecting a biological marker of disease, for example, integrin alpha 10 in the present invention, or diagnosing a disease, for example, by analyzing the expression of integrin alpha 10 in the present invention, may include comparing the analyzed tissue with healthy, non-malignant, or unaffected tissue. For example, a breast cancer sample can be compared with an unaffected area of ​​the same tissue sample or a healthy breast tissue sample. A brain cancer sample and / or a sample from a cancer of the CNS can be compared with an unaffected area of ​​the same tissue sample or a healthy brain or CNS tissue sample. A lung cancer sample can be compared with an unaffected area of ​​the same tissue sample or a healthy lung tissue sample. A prostate cancer sample can be compared with an unaffected area of ​​the same tissue sample or a healthy prostate tissue sample. A pancreatic cancer sample can be compared with an unaffected area of ​​the same tissue sample or a healthy pancreatic tissue sample. A sarcoma sample can be compared with an unaffected area of ​​the same tissue sample or a healthy connective tissue sample.

[0328] In the process of diagnosing cancer, one of skill in the art will appreciate the possible utility of comparing levels of integrin alphal0 polypeptide or polynucleotide in cancer cells to reference cells.

[0329] In some embodiments, the diagnosed cancer comprises cells that exhibit equal or higher levels of i) integrin alpha 10 antigen, or ii) polynucleotide transcripts, observed in healthy and / or benign tissue of the same type.

[0330] In some embodiments, the diagnosed cancer comprises cells that exhibit equal or higher levels of i) integrin alpha 10 antigen, or ii) polynucleotide transcripts observed in less abundant cancer types of the same tissue type compared to the diagnosed cancer type.

[0331] In some embodiments, the diagnosed cancer comprises cells that exhibit equal or greater expression levels of the integrin alpha 10 antigen.

[0332] Those skilled in the art will understand that reference cell lines can be used for standardized diagnostic procedures due to the genotypic and phenotypic stability of these cell lines compared to primary cells. Reference cell lines may be established from healthy tissue or cancer tissue, and the cancer may be a more aggressive or less aggressive cancer type depending on its tendency to grow and metastasize. Established cell lines need to be immortalized so that they can grow in cell culture. Non-malignant cell lines as used herein are understood to be established cell lines that do not show signs of malignancy and whose phenotype is similar to healthy tissue cells.

[0333] In some embodiments, the reference cell line used in the diagnostic procedure is derived from healthy tissue, e.g. selected from the group consisting of a reference cell line derived from healthy breast tissue, a reference cell line derived from healthy prostate tissue, a reference cell line derived from healthy lung tissue, a reference cell line derived from healthy pancreatic tissue, and a reference cell line derived from healthy connective tissue.

[0334] In some embodiments, the reference cell line used during the breast cancer diagnostic procedure is derived from healthy tissue.

[0335] Another aspect of the present disclosure relates to a method for detection and / or diagnosis of a cancer morphology in a subject, the method comprising: a. Providing tissue suspected of containing cancer cells of a subject; b. analyzing the presence of an antigen comprising an integrin alpha 10 polypeptide or a fragment thereof in the tissue; c. determining the expression level of integrin alpha 10 antigen; comparing the expression level determined by dc with a control level, said control level being the average expression level of the antigen observed in healthy and / or benign tissue of the same type; and comparing, a higher expression level of the antigen than the control level indicates the presence of a cancer form in the sample, said cancer form being selected from the group consisting of breast cancer, brain cancer, cancer of the CNS, lung cancer, prostate cancer, pancreatic cancer, skin cancer, lymphoma, and sarcoma, or any cancer type listed under "clinical conditions" herein, or a metastasis of any one of said forms; Thereby, a form of cancer is diagnosed in the subject.

[0336] Another aspect of the present disclosure relates to a method for diagnosis of a cancer morphology in a subject, the method comprising: a. Providing tissue suspected of containing cancer cells of a subject; b. Analyzing the presence of one or more cells having a cancerous morphology in the tissue; and c. Analyzing the presence of an antigen comprising an integrin alpha 10 polypeptide or a fragment thereof in the sample; Optionally, determining the expression level of integrin alpha 10 antigen; The presence of one or more cells having a cancer morphology in combination with expression of the integrin alpha 10 antigen indicates the presence of a cancer morphology in the sample, said cancer morphology being selected from the group consisting of breast cancer, brain cancer, cancer of the CNS, lung cancer, prostate cancer, pancreatic cancer, skin cancer, lymphoma, and sarcoma, or any cancer type listed herein under "clinical conditions", or a metastasis of any one of said cancer morphologies; Thereby, a form of cancer is diagnosed in the subject.

[0337] In some embodiments, analyzing the presence of the antigen comprising integrin alpha 10 polypeptide or a fragment thereof in step b comprises contacting the tissue suspected of containing cancer cells with the composition of the present disclosure.For example, in some embodiments, the tissue suspected of containing cancer cells may be contacted with a composition comprising or consisting of an antibody having specificity for integrin alpha 10 polypeptide.

[0338] In addition to analyzing the presence of an antigen comprising an integrin alpha 10 polypeptide or a fragment thereof, and / or the presence of a polynucleotide transcript encoding an integrin alpha 10 polypeptide or a fragment or variant thereof in a sample, and determining whether the expression level of integrin alpha 10 is higher than a control level, the method for diagnosing cancer in a subject and / or the method for detecting cancer cells in a subject may further comprise a step of morphologically characterizing the sample as containing cancer cells belonging to a cancer, said cancer form being selected from the group consisting of breast cancer, brain cancer, cancer of the CNS, lung cancer, prostate cancer, pancreatic cancer, skin cancer, lymphoma, and sarcoma, or any cancer type listed under "clinical conditions" herein, or a metastasis of any one of said cancer forms.

[0339] In another aspect, the disclosure relates to a method for detecting cancer cells in a subject, the method comprising: a. providing tissue suspected of containing cancer cells from said subject; b. Analyzing the presence of one or more cells having a cancerous morphology in the tissue; c. analyzing the tissue for the presence of an antigen comprising an integrin alpha 10 polypeptide or a fragment thereof; d. Optionally, determining the expression level of integrin alpha 10 antigen; The presence of one or more cells having a cancer morphology in combination with expression of the integrin alpha 10 antigen indicates the presence of a cancer morphology in the subject, said cancer morphology being selected from the group consisting of breast cancer, brain cancer, cancer of the CNS, lung cancer, prostate cancer, pancreatic cancer, skin cancer, lymphoma, and sarcoma, or any cancer type listed under "clinical conditions" herein, or a metastasis of any one of said cancer morphologies.

[0340] Another aspect of the disclosure relates to a method for determining a prognosis of a cancer type for a subject, the cancer type being selected from the group consisting of breast cancer, brain cancer, cancer of the CNS, lung cancer, prostate cancer, pancreatic cancer, skin cancer, lymphoma and sarcoma, or a metastasis of any one of the cancer types, the method comprising: a. Providing a subject's cancer tumor tissue; b. analyzing the sample for the presence of an antigen comprising an integrin alpha 10 polypeptide or a fragment thereof; c. determining the expression level of integrin alpha 10 antigen; comparing the expression level determined by dc with a control level, said control level being the average expression level of integrin alpha 10 antigen observed in healthy and / or benign tissue of the same tissue type as the sample; e. Determining the prognosis of the cancer to be unfavorable if the expression level of the antigen is higher than the control level.

[0341] Another aspect of the present disclosure relates to a method for determining a prognosis of cancer for a subject, the method comprising: a. Providing a subject's cancer tumor tissue; b. Analyzing the presence of one or more cells having a cancerous morphology in the tissue; and c. Analyzing the presence of an antigen comprising an integrin alpha 10 polypeptide or a fragment thereof in the sample; d. Optionally, determining the expression level of integrin alpha 10 antigen; and e. determining an unfavorable prognosis of the cancer morphology when one or more cells having the cancer morphology are present in the tissue in combination with expression of the integrin alpha 10 antigen; The cancer is selected from the group consisting of breast cancer, brain cancer, cancer of the CNS, lung cancer, prostate cancer, pancreatic cancer, skin cancer, lymphoma and sarcoma, or a metastasis of any one of the aforementioned cancers.

[0342] The method for determining the prognosis of a subject's cancer can be applied to any one of the cancer forms described herein. See, for example, the section "Clinical Status."

[0343] In some embodiments, the present disclosure relates to methods for determining a prognosis of a form of cancer in a subject, wherein the prognosis is overall survival or recurrence-free survival.

[0344] In some embodiments, the methods disclosed herein target antigens comprising an integrin alphal0 polypeptide or a fragment thereof expressed on the surface of a cell.

[0345] The methods of the present disclosure can be performed in vivo or in vitro.

[0346] In some embodiments, the methods disclosed herein for detecting cancer cells in a subject are performed in vitro. In some embodiments, the methods disclosed herein for diagnosing cancer in a subject are performed in vitro and the tissue is a tissue sample obtained from the subject.

[0347] In some embodiments, the methods disclosed herein for diagnosing cancer in a subject are performed in vivo, hi some embodiments, the methods disclosed herein for diagnosing cancer in a subject are performed in vitro and the tissue is a tissue sample obtained from the subject.

[0348] In some embodiments, the methods disclosed herein for determining the prognosis of cancer in a subject are performed in vivo. In some embodiments, the methods disclosed herein for determining the prognosis of cancer in a subject are performed in vitro and the tissue is a tissue sample obtained from the subject.

[0349] In some embodiments, the step of analyzing for the presence of an antigen comprising an integrin alphal0 polypeptide or a fragment thereof in the sample comprises imaging the tissue and / or tissue sample.

[0350] In some embodiments, determining the expression level of the integrin alpha 10 antigen comprises imaging the tissue and / or tissue sample, hi some embodiments, determining the expression level of the integrin alpha 10 antigen comprises imaging the tissue in vivo, such as during open brain surgery.

[0351] Imaging can be performed, for example, by administering to the subject and / or tissue sample a labeled moiety capable of binding to an antigen comprising an integrin alphal0 polypeptide or a fragment thereof. For example, imaging can be performed by administering to the subject and / or tissue sample a labeled integrin alphal0 antibody as defined herein.

[0352] In some embodiments, integrin alpha10 antibody is administered as part of antibody-drug conjugate (ADC). In ADC, antibody is linked to a moiety, for example, a disease-modifying drug or toxin. When specifically bound to integrin alpha10 polypeptide, ADC is internalized in the cell, thereby delivering the moiety into the cell.

[0353] In some embodiments, the antibody included in the composition for use in the diagnosis and / or treatment of a form of cancer disclosed herein is an antibody fragment.

[0354] In some embodiments, an antibody that specifically binds to an integrin alphal0 polypeptide or a fragment thereof according to the present disclosure may be conjugated to a moiety, such as an additional moiety. This conjugation may improve and facilitate both the treatment and diagnosis of cancer forms. EXAMPLES

[0355] Example 1: Antibody Production / Humanization 1.1 Design of composite human antibody variable regions Objective: To generate human antibody sequence segments to create humanized antibody variants.

[0356] Materials and Methods: A structural model of the V-region of the murine antibody mAb365 was generated. Based on the structural analysis, humanized variant sequences likely to be essential for the binding properties of the antibody were selected and analyzed in silico. Human sequence segments were identified for the CDR regions and regions outside the CDRs. The selected sequence segments were assembled to generate fully humanized V-region sequences that lacked or had reduced important T-cell epitopes to avoid immunogenicity.

[0357] Results: This design resulted in six heavy chain sequences (VH1-VH6) and five light chain sequences (VK1-VK5) that were used for gene synthesis and expression.

[0358] Mammalian cells (Table 1). The VH4 variants, which are part of the CDR4 region, were excluded due to the risk that changes in this region may affect the validation of the CDRs. Five VHs were combined with five VKs to generate 25 different humanized variants.

[0359] Conclusion: 25 humanized variants and 1 chimeric variant were generated.

[0360] [Table 1]

[0361] 1.2 Construction and transient expression of chimeric IgG1 and humanized IgG1 variants Objective: To generate small batches of antibodies for lead candidate selection.

[0362] Materials and Methods: The chimeric Tar-Ab0 sequence and humanized variants were used to express IgG1 antibodies in CHO cells. Culture supernatants were harvested 6 days after transfection and antibody concentrations were measured.

[0363] Results and conclusions: 17 different variants showed improved yields compared to the chimeric antibody (TAR-Ab0) (data not shown). TAR-Ab23 had a 3.9-fold better yield than the chimeric antibody.

[0364] 1. Selection of 35 potential leads Objective: To select 5 lead candidates from 25 humanized variants based on binding ability.

[0365] Materials and Methods: Antibody binding and specificity was tested using the mouse myoblast cell line C2C12 overexpressing either human integrin alpha10 beta1 vector (C2C12apha10) or human integrin alpha11 beta1 (C2C12alpha11). C2C12 cells were incubated with 1 μg / ml integrin alpha10 antibody for 30 min followed by secondary antibody for 30 min (100000 cells / sample) and then binding was analyzed by flow cytometry.

[0366] Results: The results showed that the chimeric form of TAR-Ab0, as well as the humanized variants TAR-Ab3, TAR-Ab8, TAR-Ab9, TAR-Ab13, TAR-Ab14, and TAR-Ab23, showed the best binding affinity to C2C12alpha10 cells (data not shown). None of the antibodies showed binding to the control C2C12alpha11 cells (data not shown). Furthermore, the risk analysis data from the sequence design process suggests that TAR-Ab3 has the highest risk score (data not shown). Therefore, variants TAR-Ab8, TAR-Ab9, TAR-Ab13, TAR-Ab14, and TAR-Ab23 were selected for further functional testing.

[0367] Conclusion: Five humanized antibody lead candidates that specifically bind to integrin alpha10 beta1 were selected: TAR-Ab8, TAR-Ab9, TAR-Ab13, TAR-Ab14, and TAR-23.

[0368] 1. Functional characterization of 45 antibody lead candidates Objective: To investigate the function-blocking effects of five lead candidates on the proliferation, adhesion, and migration of integrin alpha10 beta1-expressing cells.

[0369] Materials and Methods: The mouse myoblast cell line C2C12 overexpressing integrin alpha10 beta1 (C2C12 alpha10) was used for functional blocking studies.

[0370] Cell proliferation C2C12 alpha10 cells mixed with integrin alpha10 antibody at a final concentration of 5 μg / ml were seeded in 96-well plates (10000 cells per well) precoated with collagen type I. After 24 h of treatment with the antibody, bromodeoxyuridine (BrdU) was added for 2 h and proliferation was measured using the Cell Proliferation ELISA BrdU kit (Roche Diagnostics GmbH) according to the manufacturer's instructions.

[0371] cell adhesion Culture plates (48 wells) were coated overnight with type IV collagen (10 μg / ml) or bovine serum albumin (BSA) as a control. Prior to the experiment, the plates were incubated with 0.25% BSA for 30 min at 37°C to block non-specific binding. BT549 cells were pre-incubated for 20 min in the presence or absence of integrin alpha 10 antibody (5 μg / ml) and then allowed to attach to the coated dishes for 60 min at 37°C. Non-adherent cells were washed off, and adherent cells were fixed with ethanol and stained with 0.09% crystal violet. The absorbed dye was then extracted with 10% acetic acid and quantified by optical density (OD) at 590 nm.

[0372] Migration in 3D collagen gels Collagen gel solution was made by mixing type I collagen (6 mg / ml C2124, Sigma Aldrich) with culture medium to a final concentration of 1.2 mg / ml. Collagen gel solution (100 μl volume) was added to a 96-well plate and incubated at 37 °C for 15 min. Spheroids of C2C12 alpha 10 cells were then pipetted into the collagen solution, and the collagen-spheroid solution was allowed to polymerize at 37 °C for 60–90 min. After polymerization, 100 μl of culture medium containing 10 μg / ml of integrin alpha 10 antibody or control antibody was added to the well to allow the collagen gel to float in the antibody solution. Migration of C2C12 alpha 10 cells from the spheroids into the collagen gel was monitored using an inverted illuminated microscope, and pictures were taken at different time points for quantification.

[0373] Results: The results showed that all five antibody lead candidates significantly reduced cell proliferation (Figure 1) and adhesion (Figure 2) of C2C12 alpha 10 cells, while the isotype control antibody showed no inhibitory effect on C2C12 alpha 10 cells (Figures 1-2). In the 3D migration assay, all humanized variants except TAR-Ab14 and mAb365 caused a significant decrease in cell migration (Figure 3).

[0374] Conclusions: All five humanized antibody variants exhibit potent function-blocking properties in proliferation, adhesion, and migration assays.

[0375] Example 2: Thermal stability analysis Objective: To rank the top five humanized antibody variants based on stability.

[0376] Materials and Methods: Thermal stability analysis of humanized antibody variants was performed using the Uncle™ biostability platform and software. Samples of each variant were formulated in PBS and Sypro Orange at a final concentration of 0.5 mg / ml. Samples were subjected to a thermal gradient from 25 to 95°C with a ramp rate of 0.3°C / min and excitation at 473 nm. Protein aggregation can be detected by monitoring static light scattering (SLS) at 473 nm, and T was determined from the resulting SLS profile. agg (onset of aggregation) was calculated.

[0377] Results: All humanized antibody variants showed good pharmaceutical properties including high thermal stability and low aggregation tendency, which facilitates manufacturing and storage and suggests a long serum half-life. All humanized variants showed higher calculated melting temperatures (Tm) than the chimeric antibody (Tar-Ab0). The most thermally stable variants were TAR-Ab23, TAR-Ab13, and TAR-Ab8 (Table 4). The thermal stability of TAR-Ab23 was 17.4°C higher than the chimeric variant.

[0378] Conclusion: Thermal stability was higher for all five selected humanized antibody variants compared to the chimeric antibody variant, supporting the therapeutic use of humanized antibodies.

[0379] [Table 2]

[0380] Example 3: Isoelectric point Objective: To determine the isoelectric point (Pl) of humanized antibody variants compared to chimeric antibodies.

[0381] Materials and Methods: The isoelectric point (pI) of an antibody is the pH at which the antibody has no net charge, and its value depends on the charged amino acids contained in the antibody. The isoelectric points of antibody variants were calculated based on DNA / protein sequence analysis software.

[0382] Results: Analysis shows that TAR-Ab23 has a pl of 8.121 and the chimeric antibody Tar-Ab0 has a pl of 7.83. This is an advantage as a higher pI is associated with a proportionally higher drug exposure in the tumor, thus improving the chances of inducing a positive clinical effect.

[0383] Conclusion: TAR-Ab23 has a higher isoelectric point than chimeric antibodies, which supports a higher therapeutic potential. Therapeutic antibodies often have a pl higher than the pH of the biophase. Tumors are usually acidic due to the formation of lactic acid. As a result, antibodies with a high PI generally have an improved tumor to blood ratio compared to antibodies with a low PI. Therefore, a high PI is considered an advantage of therapeutic antibodies for cancer therapy.

[0384] Example 4: Binding affinity of TAR-Ab23 to integrin alpha10beta1 Objective: To determine the integrin alpha10 binding specificity and affinity of TAR-Ab23 compared to mAb365.

[0385] Materials and Methods: Integrin alpha10 beta1 overexpressing cells (C2C12 alpha10), human patient-derived glioblastoma cells (U3046MG), and triple-negative breast cancer cells (BT549) were used. Cells were incubated with different concentrations (0.1, 1, 10, 100, 1000 nM) of humanized (TAR-Ab23) or mouse (mAb365) antibodies for 30 min at 4°C, and then binding was analyzed by flow cytometry.

[0386] Results: Both TAR-Ab23 and mAb365 have good binding affinity to all three cell lines (C2C12 alpha 10, U3046MG, and BT549) (Figure 4A-C). However, TAR-Ab23 showed higher binding to cells compared to mAb365, especially at low concentrations, e.g., 0.1 nM (Figure 4D). Furthermore, the binding capacity of the murine antibody mAb365 decreased at higher concentrations (above 100 nM), which may be due to aggregation, whereas the binding of the humanized antibody TAR-Ab23 remained high (Figure 4).

[0387] Conclusion: These findings indicated that TAR-Ab23 has a higher binding affinity than mAb365 and that TAR-Ab23 is more stable than mAb365 at high concentrations (>100 nM).

[0388] Example 5: Specific binding of TAR-Ab23 to TNBC tissues Objective: To demonstrate the specific binding of TAR-Ab23 to integrin alpha10 beta1 on cancer cells in primary TNBC tissues.

[0389] Materials and Methods: Triple-negative human breast cancer tissues consisting of breast adenocarcinoma, ductal carcinoma, and metastases were examined for immunohistochemical staining of integrin alpha 10. Sections were stained with humanized TAR-Ab23 antibody, mouse mAb365 antibody, and isotype control antibodies human IgG1 and mouse IgG2a (all antibodies at 3 μg / ml).

[0390] Results: The TAR-Ab23 antibody showed specific and strong staining of cancer cells in TNBC tissue. In contrast, adjacent normal tissue breast ducts showed low or no TAR-Ab23 staining. The mAb365 antibody showed non-specific nuclear staining in TNBC in addition to the expected integrin alpha10 staining of TNBC cells. The isotype control antibody did not show any detectable staining.

[0391] Conclusion: TAR-Ab23 can be used to detect and target integrin alpha10beta1 in tissues due to its highly specific binding to the integrin subunit alpha10. On the other hand, the antibody mAb365 showed non-specific nuclear staining of cells both within the tumor and in the surrounding tissue. This supports the use of TAR-Ab23 both as a companion diagnostic tool and as a therapeutic agent in the treatment of TNBC and other forms of cancer.

[0392] Example 6: Binding competition assay between integrin alpha10 antibody humanized TAR-Ab23 and mouse mAb365 Objective: To investigate the affinity of antibodies TAR-Ab23 and mAb365 for integrin alpha10 in a quantitative competition assay using flow cytometry.

[0393] Materials and Methods: Binding competition assays of antibodies TAR-Ab23 and mAb365 were performed on triple-negative breast cancer cell line BT549, glioblastoma cell line U3054MG, and C2C12alpha10 cells. Antibodies were added simultaneously to cells at a concentration of 0.1 nM. After 30 min of incubation, secondary antibodies were added for another 30 min. Donkey anti-human Alexa488 was used as secondary antibody against human antibody TAR-Ab23. Donkey anti-mouse 20Alexa647 was used as secondary antibody against mouse antibody mAb365. Antibody binding was analyzed by flow cytometry.

[0394] Results: Competition assays showed that TAR-Ab23 bound to the majority of C2C12alpha10 cells (87.6%) and BT549 cells (87.3%), whereas mAb365 bound to only 29.1% of C2C12alpha10 cells and 22.9% of BT549 cells (Figure 5). Furthermore, TAR-Ab23 did not completely compete for the binding of mAb365, suggesting an overlap in the binding epitopes between the two antibodies.

[0395] Conclusion: TAR-Ab23 had more than 3-fold higher binding affinity to integrin alpha10 than mAb365, and also bound faster to integrin alpha10 antigen compared to mAb365.

[0396] Example 7: Functional blocking properties of antibody TAR-Ab23 in vitro. Inhibition of adhesion, proliferation, and migration following blockade of integrin alpha10beta1 with TAR-Ab23 7.1 Adhesion Objective: To investigate the effect of TAR-Ab23 on the adhesion of TNBC cells compared with mAb365.

[0397] Materials and Methods: Culture plates (48 wells) were coated overnight with type IV collagen (10 μg / ml) or bovine serum albumin (BSA) as a control. Prior to the experiment, the plates were incubated with 0.25% BSA for 30 min at 37°C to block non-specific binding. BT549 cells were pre-incubated for 20 min in the presence or absence of integrin alpha 10 antibody (5 μg / ml) and then allowed to attach to the coated dishes for 60 min at 37°C. Non-adherent cells were washed off, and adherent cells were fixed with ethanol and stained with 0.09% crystal violet. The absorbed dye was then extracted with 10% acetic acid and quantified by optical density (OD) at 590 nm.

[0398] Results: TAR-Ab23 significantly inhibited the adhesion of BT549 cells to collagen IV-coated dishes (86% reduction). The effect of TAR-Ab23 was more pronounced than that of mAb365 (70% reduction) (Figure 6A).

[0399] Conclusions: TAR-Ab23 is more efficient than mAb365 in inhibiting integrin alpha 10-dependent adhesion to the extracellular matrix component type IV collagen.

[0400] 7.2 Proliferation Objective: To investigate the effect of TAR-Ab23 on the proliferation of TNBC cells compared with mAb365.

[0401] Materials and Methods: BT549 cells were seeded in 96-well plates coated with type IV collagen and treated with integrin alpha 10 or control antibodies at a final concentration of 5 μg / ml. After 48 h of antibody treatment, bromodeoxyuridine (BrdU) was added for 3 h and proliferation (BrdU incorporation into DNA) was measured using the Cell Proliferation ELISA BrdU kit (Roche Diagnostics GmbH).

[0402] Results: Treatment of BT549 cells with the integrin alpha10 antibodies TAR-Ab23 and mAb365 reduced cell proliferation compared to treatment with the respective control antibodies (Th301 and mIgG2a; Figure 6B). However, TAR-Ab23 inhibited cell proliferation more efficiently than mAb365.

[0403] Conclusion: TAR-Ab23 has an inhibitory effect on TNBC cells and is more efficient compared with mAb365.

[0404] 7.33D migration Objective: To investigate the effect of TAR-Ab23 on 3D migration of TNBC cells in collagen gel compared with mAb365.

[0405] Materials and Methods: Collagen gel solution was made by mixing type I collagen (6 mg / ml C2124, Sigma Aldrich) with culture medium to a final concentration of 1.2 mg / ml. Collagen gel solution (100 μl) was added to a 96-well plate and incubated at 37 °C for 15 min. BT549 cell spheroids were then pipetted into the collagen solution, and the collagen-spheroid solution was allowed to polymerize at 37 °C for 60-90 min. After polymerization, 100 μl of culture medium containing 10 μg / ml of integrin alpha 10 antibody or control antibody was added to the well to allow the collagen gel to float in the antibody solution. Migration of BT549 cells from the spheroids into the collagen gel was monitored and photographed at different time points for quantification using an inverted illuminated microscope.

[0406] Results: Both antibodies TAR-Ab23 and mAb365 inhibited the migration of BT549 cells, whereas control antibodies (Th301 or mIgG2a) had no effect (Figure 6C). However, the inhibitory effect of TAR-Ab23 was more pronounced than that of mouse mAb365. Compared to the control antibodies, TAR-Ab23 reduced the migration of BT549 cells in collagen gel by 57%, whereas mAb365 reduced migration by 45% (Figure 6C).

[0407] Conclusion: These results suggest that TAR-Ab23 inhibits integrin alpha10-mediated cell migration more effectively than mAb365.

[0408] The in vitro function blocking properties of the integrin alpha10 antibody indicate that TAR-Ab23 can potently block anchorage-dependent cellular functions such as adhesion, proliferation, and migration.

[0409] Example 8: Function-blocking properties of antibody TAR-Ab23 in vivo. Objective: To investigate the treatment effect of TAR-Ab23 on the metastasis of triple-negative breast cancer cell line MDA-MB-231.

[0410] Materials and Methods: Luciferase / GFP-labeled MDA-MB-231 cells (2 × 10 6 Cells) were injected intravenously into 4-week-old NMRI-nu immunodeficient mice (Janvier, France) via the tail vein. One day after inoculation, D-luciferin substrate was injected subcutaneously and mice were subjected to live imaging with IVIS-CT to examine tumor progression. Based on bioluminescence images (total flux readout (photons / sec)), mice were randomly classified into three different treatment groups and injected intraperitoneally with antibodies (5 mg / kg). Mice were imaged by IVIS-CT once a week and metastases were observed. Total body weight was measured every two weeks.

[0411] Results: Imaging analysis demonstrated that TAR-Ab0 treatment suppressed spontaneous metastases, including lung and liver metastases, in mice following intravenous injection of MDA-MB-231 (Figure 7).

[0412] Conclusion: The results show that TAR-Ab23 has a direct anti-metastatic effect against TNBC in a xenograft model, indicating that the cell surface molecule integrin alpha10beta1 is important for the metastasis of TNBC cells and may represent a promising target for therapeutic cancer interference.

[0413] Example 9: The activity of therapeutic antibodies is mediated, in part, by antibody-dependent cell-mediated cytotoxicity (ADCC). Objective: To investigate whether TAR-Ab23 has the ability to activate ADCC responses.

[0414] Optimized variants of TAR-Ab23 with stronger ADCC potential have been generated by reducing fucosylation. TAR-Ab23 low fucose vs. TAR-Ab23 were also used to quantify ADCC activity in gene reporter assays.

[0415] Materials and Methods: Jurkat Lucia NFAT-CD16 effector cells were passaged 2 days before the ADCC assay. The day before the assay, A204 target cells were seeded in complete medium in flat-bottom 96-well plates (SPL) based on the effector / target cell ratio according to the plate layout. On the day of the ADCC assay, cell culture medium was removed from the 96-well plates with attached target cells and 90ul of fresh test medium (exp#1-3: RPMI-1640 + 10% heat-inactivated FBS, exp#4-6: RPMI-1640 + 2% heat-inactivated low IgG FBS) was added. To this target cells, 20μl of antibody dilution series was added to each well and incubated for 1 hour. After incubation, 90μl of Jurkat-Lucia NFAT-CD16 effector cells were added according to the effector / target cell ratio (2:1). The plates were incubated for 24 hours. 20 μl of cell supernatant was transferred to a 96-well white (opaque) plate, and 50 μl of Quench-Luc was added to each well and immediately subjected to measurement in a luminometer.

[0416] Results: The ADCC activity of TAR-Ab23 (A) and TAR-Ab23 low fucose (B) was determined using Jurkat Lucia / NFAT-CD16 cells. TAR-Ab23 produced a dose-dependent signal curve compared to the control antibody by exhibiting ADCC activity in the A204 rhabdoid cell line. The ADCC activity was significantly enhanced by reducing the fucose content via afucosylation. The low fucose of TAR-mAb23 resulted in a dose-dependent ADCC response compared to the parent antibody at much lower concentrations.

[0417] Conclusion: Our data demonstrated that TAR-Ab23 induced ADCC. Furthermore, the low-fucose variant of TAR-Ab23 was shown to have higher ADCC activity.

[0418] Example 10: Antibody sequences SEQ ID NO: 1: RASKSVSTSGFSHMH Variable light chain complementarity determining region 1 (CDR-L1), constant among the five light chain humanized variants. SEQ ID NO:2: LASNLES Variable light chain complementarity determining region 2 (CDR-L2) of VK1, VK2, and VK3. SEQ ID NO:3: LGSNLES Variable light chain complementarity determining region 2 (CDR-L2) of VK4. SEQ ID NO:4: GASNLES Variable light chain complementarity determining region 2 (CDR-L2) of VK5. SEQ ID NO:5: QHSRELPRT Variable light chain complementarity determining region 3 (CDR-L3), constant among the five light chain humanized variants. SEQ ID NO: 6: DYNMD Variable heavy chain complementarity determining region 1 (CDR-H1) of VH1, VH2 and VH3. SEQ ID NO: 7: DYNMN Variable heavy chain complementarity determining region 1 (CDR-H1) of VK5. SEQ ID NO: 8: DYNMH Variable heavy chain complementarity determining region 1 (CDR-H1) of VK6. SEQ ID NO: 9: DINPNTGGTIYNQKFKG Variable heavy chain complementarity determining region 2 (CDR-H2), constant among the five heavy chain humanized variants. SEQ ID NO: 10: REDWYYFDF Variable heavy chain complementarity determining region 3 (CDR-H3), constant among the five heavy chain humanized variants. SEQ ID NO: 11 (variable light chain - VK1) DIVLTQSPATLALSPGERATLSCRASKSVSTSGFSHMHWYQQKPGQPPKLLIYLASNLESGVPSRFSFSGSGTDFTLNIHPVQEEDAATYYCQHSRELPRTFGGGGTKVEIK SEQ ID NO: 12 (variable light chain - VK2) DIVLTQSPATLSLSPGERATLSCRASKSVSTSGFSHMHWYQQKPGQPPKLLIYLASNLESGVPSRFSFSGSGTDFTLTISSLQPEDVATYYCQHSRELPRTFGGGGTKVEIK SEQ ID NO: 13 (variable light chain - VK3) DIVLTQSPATLLSPGERATLSCRASKSVSTSGFSHMHWYQQKPGQPPKLLIYLASNLESGVPSRFSGSGSGTDFTLTISSLQPEDVATYYCQHSRELPRTFGGGGTKVEIK SEQ ID NO: 14 (Variable light chain - VK4) DIVLTQSPATLSLSPGERATLSCRASKSVSTSGFSHMHWYQQKPGQPPKLLIYLGSNLESGVPSRFSFSGSGTDFTLTISSLQPEDVATYYCQHSRELPRTFGGGGTKVEIK SEQ ID NO: 15 (variable light chain - VK5) DIVLTQSPATLSLSPGERATLSCRASKSVSTSGFSHMHWYQQKPGQPPKLLIYGASNLESGVPSRFSFSGSGTDFTLTI SSLQPEDVATYYCQHSRELPRTFGGGGTKVEIK SEQ ID NO: 16 (variable heavy chain - VH1) EVQLVQSGPELKKPGASVKVPCKASGYTFTDYNMDWVRQPHGKGLEWIGDINPNTGGTIYNQKFKGRATITVDKSTSTAYMELSSLRSEDTSVYYCARREDWYYFDFWGQGTTVTVSS SEQ ID NO: 17 (variable heavy chain - VH2) EVQLVQSGAEVKKPGASVKVSCKASGYTFTDYNMDWVRQPPGKGLEWIGDINPNTGGTIYNQKFKGRATITVDKSTSTAYMELSSLRSEDTSVYYCARREDWYYFDFWGQGTTVTVSS SEQ ID NO: 18 (variable heavy chain - VH3) EVQLVQSGAEVKKPGASVKVSCKASGYTFTDYNMDWVRQPPGKGLEWIGDINPNTGGTIYNQKFKGRATITVDKSTSTAYMELSSLRSEDTAVYYCARREDWYYFDFWGQGTTVTVSS SEQ ID NO: 19 (variable heavy chain - VH5) EVQLVQSGAEVKKPGASVKVSCKASGYTFTDYNMNWVRQPPGKGLEWIGDINPNTGGTIYNQKFKGRATITVDKSTSTAYMELSSLRSEDTAVYYCARREDWYYFDFWGQGTTVTVSS Accession number 20 (Variable heavy chain - VH6) EVQLVQSGAEVKKPGASVKVSCKASGYTFTDYNMHWVRQPPGKGLEWIGDINPNTGGTIYNQKFKGRATITVDKSTSTAYMELSSLRSEDTAVYYCARREDWYYFDFWGQGTTVTVSS References Bengtsson T, Aszodi A, Nicolae C, Hunziker EB, Lundgren - Akerlund E, Fassler R. 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Chmielarska Masoumi K,Huang X,Sime W,Mirkov A,Munksgaard Thoren M,Massoumi R,Lundgren-Akerlund E,Integrin α10-Antibodies Reduce Glioblastoma Tumor Growth and Cell Migration,Cancers (Basel),2021;13(5):1184. Foote J,Winter G,Antibody framework residues affecting the conformation of the hypervariable loops,J Mol Biol,1992 Mar 20;224(2):487-99. Frith JE,Mills RJ,Hudson JE,Cooper-White JJ,Tailored integrin-extracellular matrix interactions to direct human mesenchymal stem cell differentiation,Stem Cells Dev,35 2012;21(13):2442-56. Gullberg DE,Lundgren-Akerlund E,Collagen-binding I domain integrins--what do they do? Prog Histochem Cytochem,2002;37(1):3-54. Heino J,The collagen receptor integrins have distinct ligand recognition and signalling functions,Matrix Biology,2000;19:319-323. Hering TM,Regulation of chondrocyte gene expression,Frontiers in Bioscience. 1999;4:743-761. Hohenester E,Signalling complexes at the cell-matrix interface,Curr Opin Struct Biol,2014;29:10-6. Jones PT,Dear PH,Foote J,Neuberger MS,Winter G,Replacing the complementarity-determining regions in a human antibody with those from a mouse,Nature,1986;4;321(6069):522-5. Kyostila K,Lappalainen AK,Lohi H,Canine chondrodysplasia caused by a truncating mutation in collagen-binding integrin alpha subunit 10,PLoS One。2013;8(9):e75621. Lundgren-Akerlund E,and Aszodi A,Integrin alpha10beta1: a collagen receptor critical in skeletal development,Adv,Exp,Med,Biol,2014;819:61-71. Munksgaard Thoren M,Chmielarska Masoumi K,Krona C,Huang X,Kundu S,Schmidt L,Forsberg-Nilsson K,Floyd Keep M,Englund E,Nelander S,Holmqvist B,Lundgren-Akerlund E,Integrin α10,a Novel Therapeutic Target in Glioblastoma,Regulates Cell Migration,Proliferation,and Survival,Cancers (Basel),2019;25;11(4):587. Quenn C,Schneider WP,Selick HE,Payne PW,Landolfi N,Duncan JF,Avdalovic NM,Levitt M,Junghans RP,and Waldmann T,A humanized antibody that binds to the interleukin 2 receptor,Proc,Nati,Acad,Sci,1989;86,10029-10033. Riechmann L,Clark M,Waldmann H,Winter G,Reshaping human antibodies for therapy,Nature,1988;24;332(6162):323-7. Sanada H,Kobayashi K,Oyama K,Maru T,Nakanishi T,Umetsu M,Asano R,Kumagai I,Affinity maturation of humanized anti-epidermal growth factor receptor antibody using a modified phage-based open sandwich selection method,Sci Rep,2018;8(1):5414. Sielska M,Przanowski P,Pasierbinska M,Wojnicki K,Poleszak K,Wojtas B,Grzeganek D,Ellert-Miklaszewska A,Ku MCh,Kettenmann H,Kaminska B,Tumour-derived CSF2 / granulocyte macrophage colony stimulating factor controls myeloid cell accumulation and progression of gliomas,Br J Cancer,2020;123(3):438-448. Varas L,Ohlsson LB,Honeth G,Olsson A,Bengtsson T,Wiberg C et al,Alpha10 integrin expression is up-regulated on fibroblast growth factor-2-treated mesenchymal stem cells with improved chondrogenic differentiation potential,Stem Cells Dev 2007;16:965-978. 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Claims

1. An antibody or antigen-binding fragment thereof having binding specificity for integrin alpha 10, said antibody or antigen-binding fragment comprising: A light chain variable region comprising: a) CDR-L1 comprising the amino acid sequence of SEQ ID NO: 1; b) a CDR-L2 comprising an amino acid sequence selected from SEQ ID NO: 2 and SEQ ID NO: 4; and c) CDR-L3 comprising the amino acid sequence of SEQ ID NO: 5; the light chain variable region comprising A heavy chain variable region comprising: d) CDR-H1 comprising an amino acid sequence selected from SEQ ID NO: 7, SEQ ID NO: 6 and SEQ ID NO: 8; e) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 9; and f) CDR-H3 comprising the amino acid sequence of SEQ ID NO: 10; the heavy chain variable region comprising The antibody or antigen-binding fragment thereof, comprising:

2. the antibody or antigen-binding fragment thereof A light chain variable region comprising: a) CDR-L1 consisting of the amino acid sequence of SEQ ID NO: 1; b) a CDR-L2 consisting of an amino acid sequence selected from SEQ ID NO: 2 and SEQ ID NO: 4; and c) CDR-L3 consisting of the amino acid sequence of SEQ ID NO: 5; the light chain variable region comprising A heavy chain variable region comprising: a) CDR-H1 consisting of an amino acid sequence selected from SEQ ID NO: 7, SEQ ID NO: 6 and SEQ ID NO: 8; b) CDR-H2 consisting of the amino acid sequence of SEQ ID NO: 9, and c) CDR-H3 consisting of the amino acid sequence of SEQ ID NO: 10; the heavy chain variable region comprising Including, binds to the extracellular I domain of integrin alpha10beta1; The antibody or antigen-binding fragment of claim 1.

3. The antibody or antigen-binding fragment of claim 1, wherein the antibody or antigen-binding fragment comprises a light chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO: 13, or an amino acid sequence having at least 85% sequence identity to SEQ ID NO:

13.

4. The antibody or antigen-binding fragment of claim 1, wherein the antibody or antigen-binding fragment comprises a heavy chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO: 19, or an amino acid sequence having at least 85% sequence identity to SEQ ID NO:

19.

5. The antibody or antigen-binding fragment thereof according to claim 1, wherein the antibody or antigen-binding fragment thereof comprises a light chain variable region consisting of SEQ ID NO: 13 and a heavy chain variable region consisting of SEQ ID NO:

19.

6. An antibody or antigen-binding fragment thereof according to claim 1, comprising or consisting of an antigen-binding fragment selected from an Fv fragment, a Fab-like fragment and a domain antibody, wherein the antibody or antigen-binding fragment thereof comprises an Fc region and / or an intact antibody.

7. The Fv fragment is selected from a single chain Fv and a disulfide-linked Fv, and the Fab-like fragment is selected from a Fab fragment, a Fab' fragment, and a F(ab) fragment. ２ fragments, and the domain antibody is selected from a single V Ｈ Variable domains and V Ｌ The antibody or antigen-binding fragment thereof of claim 6, wherein the antibody or antigen-binding fragment is selected from the variable domain.

8. The antibody or antigen-binding fragment thereof of claim 1 conjugated to an additional moiety, wherein the additional moiety comprises a cytotoxic moiety selected from a toxin, a chemotherapeutic agent, and a radioactive agent, or a combination thereof.

9. The antibody or antigen-binding fragment thereof of claim 8, wherein the toxin is selected from doxorubicin, a doxorubicin derivative, and amanitin.

10. The antibody or antigen-binding fragment thereof according to claim 9, wherein the doxorubicin derivative is 3'-deamino-3"-4'-anhydro-[2"(S)-methoxy-3"(R)-hydroxy-4"-morpholinyl]doxorubicin.

11. A polynucleotide encoding the antibody or antigen-binding fragment thereof described in claim 1, or a constituent polypeptide chain thereof.

12. A recombinant host cell comprising the polynucleotide of claim 11.

13. A pharmaceutical composition for use in medicine, comprising an antibody or antigen-binding fragment thereof described in any one of claims 1 to 10, a polynucleotide described in claim 11, and / or a recombinant host cell described in claim 12.

14. A pharmaceutical composition for use in the prevention, treatment, mitigation, detection and / or diagnosis of a disease or disorder susceptible to treatment with an integrin alpha10 inhibitor and / or a disease or disorder associated with cells expressing integrin alpha10 beta1, comprising an antibody or antigen-binding fragment thereof according to any one of claims 1 to 10, a polynucleotide according to claim 11, and / or a recombinant host cell according to claim 12.

15. The pharmaceutical composition of claim 14, wherein the disease or disorder is a neoplastic disease or disorder.

16. 16. The pharmaceutical composition of claim 15, wherein the neoplastic disease or disorder is a solid tumor selected from the group consisting of prostate cancer, breast cancer, brain cancer, cancer of the CNS, lung cancer, melanoma, pancreatic cancer, skin cancer, lymphoma, sarcoma, or metastasis of any one of the foregoing cancer forms.

17. 1. An in vitro method for the detection of cells expressing integrin alpha10 beta1 in a subject, said method comprising: (a) providing a sample of cells from a subject to be tested; (b) extracting and / or purifying cells present in said sample; (c) contacting the antibody or antigen-binding fragment thereof according to any one of claims 1 to 8 with cells present in the sample; (d) determining whether the antibody or antigen-binding fragment thereof binds to the cell; Including, Binding of the antibody or antigen-binding fragment thereof to the cell indicates the presence of a disease or disorder associated with cells expressing integrin alpha 10 in the tissue of the subject. The method.

18. The method of claim 17, wherein the sample of cells derived from the subject being tested is a biopsy tissue or blood sample.

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