Anti-uPAR antibodies and uses thereof

Abstract

The presently disclosed subject matter provides antibodies or antigen-binding fragments thereof that bind to uPAR, and methods of using such antibodies or antigen-binding fragments thereof.

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to U.S. Provisional Application No. 63 / 209,941, filed June 11, 2021, the contents of which are incorporated by reference in their entirety and priority is claimed.

[0002] Sequence Listing This application contains a Sequence Listing that was submitted in ASCII format via EFS-Web and is incorporated herein by reference in its entirety. The ASCII copy, created on Jun. 10, 2022, is named 072734.1362_ST25.txt and is 31,984 bytes in size.

[0003] The subject matter disclosed herein relates to antibodies that bind to uPAR and methods of using such antibodies. [Background technology]

[0004] uPAR is associated with tumor growth or metastasis in a variety of different types of cancer, including breast cancer, endometrial cancer, ovarian cancer, colon cancer, rectal cancer, lung cancer, gastric cancer, prostate cancer, renal cancer, pancreatic cancer, rectal cancer, acute lymphoblastic leukemia (ALL), chronic lymphocytic leukemia (CLL), and acute myeloid leukemia (AML). It also plays a role in aging, such as its association with aging-related diseases associated with aging. It can also regulate immune responses and cell-matrix interactions, promoting tumor cell proliferation and awakening from dormancy.

[0005] Given the important role of uPAR in a variety of diseases or disorders, antibodies that recognize uPAR and methods of using such agents are desirable. Summary of the Invention [Means for solving the problem]

[0006] The presently disclosed subject matter provides antibodies or antigen-binding fragments thereof that specifically bind to uPAR, and methods of using the antibodies or antigen-binding fragments thereof.

[0007] In certain embodiments, the uPAR antibody or antigen-binding fragment thereof comprises a heavy chain variable region comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or at least about 100% homologous or identical to the amino acid sequence set forth in SEQ ID NO:25, SEQ ID NO:27, SEQ ID NO:29, SEQ ID NO:31, SEQ ID NO:47, or SEQ ID NO:55. In certain embodiments, the anti-uPAR antibody or antigen-binding fragment thereof comprises a heavy chain variable region comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or at least about 100% homologous or identical to the amino acid sequence set forth in SEQ ID NO:27.

[0008] In certain embodiments, the anti-uPAR antibody or antigen-binding fragment thereof comprises a light chain variable region comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or at least about 100% homologous or identical to the amino acid sequence set forth in SEQ ID NO:26, SEQ ID NO:28, SEQ ID NO:30, SEQ ID NO:32, SEQ ID NO:48, or SEQ ID NO:56. In certain embodiments, the anti-uPAR antibody or antigen-binding fragment thereof comprises a light chain variable region comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or at least about 100% homologous or identical to the amino acid sequence set forth in SEQ ID NO:28.

[0009] In certain embodiments, the anti-uPAR antibody or antigen-binding fragment thereof comprises: (a) a heavy chain variable region comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or at least about 100% homologous or identical to the amino acid sequence set forth in SEQ ID NO:25, SEQ ID NO:27, SEQ ID NO:29, SEQ ID NO:31, SEQ ID NO:47, or SEQ ID NO:55; and (b) a light chain variable region comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or at least about 100% homologous or identical to the amino acid sequence set forth in SEQ ID NO:26, SEQ ID NO:28, SEQ ID NO:30, SEQ ID NO:32, SEQ ID NO:48, or SEQ ID NO:56.

[0010] In certain embodiments, the heavy and light chain variable regions of the anti-uPAR antibody or antigen-binding fragment thereof are (a) a heavy chain variable region comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or at least about 100% homologous or identical to the amino acid sequence set forth in SEQ ID NO:25, and a light chain variable region comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or at least about 100% homologous or identical to the amino acid sequence set forth in SEQ ID NO:26; (b) a heavy chain variable region comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or at least about 100% homologous or identical to the amino acid sequence set forth in SEQ ID NO:27, and a light chain variable region comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or at least about 100% homologous or identical to the amino acid sequence set forth in SEQ ID NO:28; (c) a heavy chain variable region comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or at least about 100% homologous or identical to the amino acid sequence set forth in SEQ ID NO:29, and a light chain variable region comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or at least about 100% homologous or identical to the amino acid sequence set forth in SEQ ID NO:30; (d) a heavy chain variable region comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or at least about 100% homologous or identical to the amino acid sequence set forth in SEQ ID NO:31, and a light chain variable region comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or at least about 100% homologous or identical to the amino acid sequence set forth in SEQ ID NO:32; (e) a heavy chain variable region comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or at least about 100% homologous or identical to the amino acid sequence set forth in SEQ ID NO:47, and a light chain variable region comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or at least about 100% homologous or identical to the amino acid sequence set forth in SEQ ID NO:48, and (f) a heavy chain variable region comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or at least about 100% homologous or identical to the amino acid sequence set forth in SEQ ID NO:55; and a light chain variable region comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or at least about 100% homologous or identical to the amino acid sequence set forth in SEQ ID NO:56.

[0011] In certain embodiments, the anti-uPAR antibody or antigen-binding fragment thereof comprises (a) a heavy chain variable region comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or at least about 100% homologous or identical to the amino acid sequence set forth in SEQ ID NO:27, and (b) a light chain variable region comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or at least about 100% homologous or identical to the amino acid sequence set forth in SEQ ID NO:28.

[0012] In certain embodiments, the anti-uPAR antibody or antigen-binding fragment thereof comprises a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO:25, SEQ ID NO:27, SEQ ID NO:29, SEQ ID NO:31, SEQ ID NO:47, or SEQ ID NO:55. In certain embodiments, the anti-uPAR antibody or antigen-binding fragment thereof comprises a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO:27. In certain embodiments, the anti-uPAR antibody or antigen-binding fragment thereof comprises a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:26, SEQ ID NO:28, SEQ ID NO:30, SEQ ID NO:32, SEQ ID NO:48, or SEQ ID NO:56. In certain embodiments, the anti-uPAR antibody or antigen-binding fragment thereof comprises a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:28.

[0013] In certain embodiments, the anti-uPAR antibody or antigen-binding fragment thereof comprises a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO:25, SEQ ID NO:27, SEQ ID NO:29, SEQ ID NO:31, SEQ ID NO:47, or SEQ ID NO:55, and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:26, SEQ ID NO:28, SEQ ID NO:30, SEQ ID NO:32, SEQ ID NO:48, or SEQ ID NO:56.

[0014] In certain embodiments, the anti-uPAR antibody or antigen-binding fragment thereof comprises a heavy chain variable region and a light chain variable region, (a) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 25, and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 26; (b) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 27, and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 28; (c) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 29, and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 30; (d) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 31, and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 32; (e) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 47, and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 48; and (f) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 55, and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 56.

[0015] In certain embodiments, the anti-uPAR antibody or antigen-binding fragment thereof comprises a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO:27, and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:28.

[0016] In certain embodiments, the anti-uPAR antibody or antigen-binding fragment thereof comprises a heavy chain variable region comprising a CDR1 domain, a CDR2 domain, and a CDR3 domain, and a light chain variable region comprising a CDR1 domain, a CDR2 domain, and a CDR3 domain, wherein the CDR3 domain of the heavy chain variable region and the light chain variable region comprises (a) a heavy chain variable region CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 3 and a conservative modification thereof, and a light chain variable region CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 6 and a conservative modification thereof; (b) a heavy chain variable region CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 9 and a conservative modification thereof, and a light chain variable region CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 12 and a conservative modification thereof; (c) a heavy chain variable region CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 15 and a conservative modification thereof, and a light chain variable region CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 18 and a conservative modification thereof; (d) a heavy chain variable region CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 21 and a conservative modification thereof, and a light chain variable region CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 24 and a conservative modification thereof; (e) a heavy chain variable region CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 43 and a conservative modification thereof, and a light chain variable region CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46 and a conservative modification thereof; and (f) a heavy chain variable region CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 51 and a conservative modification thereof, and a light chain variable region CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 54 and a conservative modification thereof.

[0017] In certain embodiments, the CDR2 domains of the heavy and light chain variable regions of the antibody or antigen-binding fragment thereof comprise: (a) a heavy chain variable region CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 2 and a conservative modification thereof, and a light chain variable region CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 5 and a conservative modification thereof; (b) a heavy chain variable region CDR2 comprising the amino acid sequence set forth in SEQ ID NO:8 and a conservative modification thereof, and a light chain variable region CDR2 comprising the amino acid sequence set forth in SEQ ID NO:11 and a conservative modification thereof; (c) a heavy chain variable region CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 14 and a conservative modification thereof, and a light chain variable region CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 17 and a conservative modification thereof; (d) a heavy chain variable region CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 20 and a conservative modification thereof, and a light chain variable region CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 23 and a conservative modification thereof; (e) a heavy chain variable region CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 42 and a conservative modification thereof, and a light chain variable region CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 45 and a conservative modification thereof; and (f) a heavy chain variable region CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 50 and a conservative modification thereof, and a light chain variable region CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 53 and a conservative modification thereof.

[0018] In certain embodiments, the CDR1 domains of the anti-uPAR heavy chain variable region and light chain variable region of the antibody or antigen-binding fragment thereof are (a) a heavy chain variable region CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 1 and a conservative modification thereof, and a light chain variable region CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 4 and a conservative modification thereof; (b) a heavy chain variable region CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 7 and a conservative modification thereof, and a light chain variable region CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 10 and a conservative modification thereof; (c) a heavy chain variable region CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 13 and a conservative modification thereof, and a light chain variable region CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 16 and a conservative modification thereof; (d) a heavy chain variable region CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 19 and a conservative modification thereof, and a light chain variable region CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 22 and a conservative modification thereof; (e) a heavy chain variable region CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 41 and a conservative modification thereof, and a light chain variable region CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 44 and a conservative modification thereof; and (f) a heavy chain variable region CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 49 and a conservative modification thereof, and a light chain variable region CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 52 and a conservative modification thereof.

[0019] In certain embodiments, one or more of the CDR sequences have up to about 5 amino acid substitutions. In certain embodiments, one or more of the CDR sequences have up to about 3 amino acid substitutions.

[0020] In certain embodiments, the anti-uPAR antibody or antigen-binding fragment thereof is (a) CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 1, CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 2, and CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 3; (b) CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 7, CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 8, and CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 9; (c) CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 13, CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 14, and CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 15; (d) CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 19, CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 20, and CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 21; (e) a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 41, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 42, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 43; or (f) a heavy chain variable region comprising CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 49, CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 50, and CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 51.

[0021] In certain embodiments, the anti-uPAR antibody or antigen-binding fragment thereof is (a) CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 4, CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 5, and CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 6; (b) CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 10, CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 11, and CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 12; (c) CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 16, CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 17, and CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 18; (d) CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 22, CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 23, and CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 24; (e) a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 44, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 45, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46; or (f) a light chain variable region comprising CDR1 having the amino acid sequence set forth in SEQ ID NO: 52, CDR2 having the amino acid sequence set forth in SEQ ID NO: 53, and CDR3 having the amino acid sequence set forth in SEQ ID NO: 54.

[0022] In certain embodiments, the anti-uPAR antibody or antigen-binding fragment thereof is (a) a heavy chain variable region comprising CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 1, CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 2, and CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 3, and a light chain variable region comprising CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 4, CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 5, and CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 6; (b) a heavy chain variable region comprising CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 7, CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 8, and CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 9, and a light chain variable region comprising CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 10, CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 11, and CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 12; (c) a heavy chain variable region comprising CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 13, CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 14, and CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 15, and a light chain variable region comprising CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 16, CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 17, and CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 18; (d) a heavy chain variable region comprising CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 19, CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 20, and CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 21, and a light chain variable region comprising CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 22, CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 23, and CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 24; (e) a heavy chain variable region comprising CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 41, CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 42, and CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 43, and a light chain variable region comprising CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 44, CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 45, and CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46; or (f) a heavy chain variable region comprising CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 49, CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 50, and CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 51, and a light chain variable region comprising CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 52, CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 53, and CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 54.

[0023] In certain embodiments, the anti-uPAR antibody or antigen-binding fragment thereof comprises a heavy chain variable region comprising CDR1 having the amino acid sequence set forth in SEQ ID NO:7, CDR2 having the amino acid sequence set forth in SEQ ID NO:8, and CDR3 having the amino acid sequence set forth in SEQ ID NO:9, and a light chain variable region comprising CDR1 having the amino acid sequence set forth in SEQ ID NO:10, CDR2 having the amino acid sequence set forth in SEQ ID NO:11, and CDR3 having the amino acid sequence set forth in SEQ ID NO:12.

[0024] In certain embodiments, the antibody sequence is in the light-heavy variable chain orientation (V L -V H ). In certain embodiments, the antibody or antigen-binding fragment thereof comprises a human variable region framework region. In certain embodiments, the antibody or antigen-binding fragment thereof is fully human or an antigen-binding fragment thereof. In certain embodiments, the antibody or antigen-binding fragment thereof is a chimeric antibody or an antigen-binding fragment thereof. In certain embodiments, the antibody or antigen-binding fragment thereof is a humanized antibody or an antigen-binding fragment thereof. In certain embodiments, the antigen-binding fragment of the antibody is a Fab, Fab', F(ab')2, a variable fragment (Fv), or a single chain variable region (scFv).

[0025] In certain embodiments, the antigen-binding fragment of the antibody or antigen-binding fragment thereof is an scFv.

[0026] Additionally, the presently disclosed subject matter provides antibodies, or antigen-binding fragments thereof, that cross-compete with any of the above-described antibodies, or antigen-binding fragments thereof, for binding to uPAR.

[0027] The presently disclosed subject matter further provides an antibody or antigen-binding fragment thereof that binds to the same epitope region on uPAR as any of the above antibodies or antigen-binding fragments thereof.

[0028] The presently disclosed subject matter also provides an immunoconjugate comprising an antibody or antigen-binding fragment thereof disclosed herein linked to a therapeutic agent, hi certain embodiments, the therapeutic agent is a drug, a cytotoxin, or a radioisotope.

[0029] The presently disclosed subject matter further provides multispecific molecules comprising an antibody or antigen-binding fragment thereof disclosed herein linked to a second functional moiety, in certain embodiments, the second functional moiety has a different binding specificity than the antibody or antigen-binding fragment thereof.

[0030] Additionally, the presently disclosed subject matter provides a composition comprising an antibody or antigen-binding fragment thereof disclosed herein, an immunoconjugate disclosed herein, or a multispecific molecule disclosed herein, hi certain embodiments, the composition is a pharmaceutical composition further comprising a pharma- ceutically acceptable carrier.

[0031] In addition, the presently disclosed subject matter provides nucleic acids encoding the antibodies or antigen-binding fragments thereof disclosed herein, vectors comprising such nucleic acid molecules, and host cells comprising such vectors.

[0032] The subject matter disclosed herein provides a method for detecting uPAR in cell, tissue or blood sample.In certain embodiments, the method includes: contacting cell, tissue or blood sample with the antibody or antigen-binding fragment thereof disclosed herein, wherein the antibody or antigen-binding fragment thereof comprises detectable label; and determining the amount of the labeled antibody or antigen-binding fragment thereof bound to cell, tissue or blood sample by measuring the amount of detectable label associated with cell, tissue or blood sample, wherein the amount of bound antibody or antigen-binding fragment thereof indicates the amount of uPAR in cell, tissue or blood sample.

[0033] The subject matter disclosed herein further provides a method for treating or alleviating a disease or disorder in a subject. In certain embodiments, the method comprises administering to a subject an antibody or an antigen-binding fragment thereof, an immunoconjugate thereof, a multispecific molecule, or a composition disclosed herein. In certain embodiments, the disease or disorder expresses uPAR. In certain embodiments, the disease or disorder is associated with overexpression of uPAR. In certain embodiments, the disease or disorder is selected from the group consisting of tumors, or aging-related pathologies, and age-related tissue loss. In certain embodiments, the disease or disorder is selected from the group consisting of pulmonary fibrosis, cardiac fibrosis, liver fibrosis, atherosclerosis, osteoarthritis, diabetes, chronic kidney disease, Alzheimer's disease, and Parkinson's disease.

[0034] In certain embodiments, the disease or disorder is an aging-associated condition, hi certain embodiments, the aging-associated condition is selected from the group consisting of pulmonary fibrosis, atherosclerosis, Alzheimer's disease, diabetes, osteoarthritis, liver fibrosis, chronic kidney disease, cardiac fibrosis, and Parkinson's disease.

[0035] In certain embodiments, the disease or disorder is a tumor. In certain embodiments, the tumor is selected from the group consisting of breast cancer (including triple-negative breast cancer), endometrial cancer, ovarian cancer, colon cancer, rectal cancer, lung cancer (e.g., non-small cell lung cancer), stomach cancer, prostate cancer, gastric cancer, renal cancer, pancreatic cancer, blood cancer, cervical cancer, head and neck cancer, liver cancer (e.g., cholangiocarcinoma, hepatocellular carcinoma, and fibrolamaellar hepatocellular carcinoma), urothelial cancer, melanoma, and brain cancer (including glioblastoma multiforme). In certain embodiments, the blood cancer is selected from the group consisting of acute lymphoblastic leukemia (ALL), chronic lymphocytic leukemia (CLL), acute myeloid leukemia (AML), myelofibrosis, polycythemia vera, myelodysplastic syndrome, and erythroleukemia. In certain embodiments, the tumor is cancer.

[0036] The subject matter disclosed herein further provides a method of increasing production of an immune-activating cytokine in response to tumor cells in a subject. In certain embodiments, the method comprises administering to the subject an antibody or antigen-binding fragment thereof, immunoconjugate thereof, multispecific molecule thereof, or composition thereof disclosed herein. In certain embodiments, the immune-activating cytokine is selected from the group consisting of granulocyte-macrophage colony-stimulating factor (GM-CSF), IFN-α, IFN-β, IFN-γ, TNF-α, IL-1, IL-2, IL-3, IL-6, IL-11, IL-7, IL-8, IL-12, IL-15, IL-21, interferon regulatory factor 7 (IRF7), CCL1, CCL2, CCL3, CCL5, CCL7, CCL8, CCL13, CCL16, CXCL1, CXCL3, CXCL5, CXCL9, CXCL10, and combinations thereof. In certain embodiments, the subject is a human.

[0037] The presently disclosed subject matter further provides a kit for treating or alleviating a disease or disorder in a subject and / or increasing the production of immune stimulating cytokines in response to tumor cells in a subject, the kit comprising an antibody or antigen-binding fragment thereof, immunoconjugate thereof, multispecific molecule thereof, or composition disclosed herein. In certain embodiments, the kit further comprises written instructions for using the antibody or antigen-binding fragment thereof, immunoconjugate thereof, multispecific molecule thereof, or composition disclosed herein for treating or alleviating a disease or disorder in a subject and / or increasing the production of immune stimulating cytokines in response to tumor cells in a subject. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0038] The presently disclosed subject matter provides anti-uPAR antibodies. Non-limiting embodiments of the disclosure are illustrated herein and by way of examples.

[0039] For purposes of clarity of disclosure, and not by way of limitation, this detailed description is divided into the following subsections. 4.1. Definition, 4.2.uPAR, 4.3. Anti-uPAR antibody, 4.4. Nucleic Acids Encoding Antibodies or Antigen-Binding Fragments 4.5. Pharmaceutical Compositions and Methods of Treatment 4.6. Diagnostic and prognostic methods; 4.7. Kits, and 4.8. EXEMPLARY EMBODIMENTS

[0040] 4.1.Definition In the following description, certain rules are followed regarding the use of terms: In general, the terms used herein are intended to be interpreted consistently with the meaning of those terms as they are known to those skilled in the art.

[0041] An "antigen binding protein" is a protein or polypeptide that contains an antigen-binding region or antigen-binding fragment, i.e., that has a strong affinity for another molecule to which it binds. Antigen binding proteins include antibodies, chimeric antigen receptors (CARs), and fusion proteins.

[0042] The terms "antibody" and "antibody" as known in the art refer to an antigen-binding protein of the immune system. The term "antibody" as referred to herein includes full-length antibodies having an antigen-binding region, and an "antigen-binding fragment" or any fragment thereof in which the "antigen-binding region" is retained, or a single chain thereof, e.g., a single-chain variable fragment (scFv). A naturally occurring "antibody" is a glycoprotein comprising at least two heavy (H) chains and two light (L) chains interconnected by disulfide bonds. Each heavy chain comprises a heavy chain variable region (herein referred to as V H Each light chain is made up of a light chain variable region (abbreviated herein as V) and a heavy chain constant region (CH). The heavy chain constant region is made up of three domains, CH1, CH2, and CH3. L (abbreviated as ) and light chain constant C L The light chain constant region consists of one domain, C L It consists of V H Area and V L The regions can be further subdivided into regions of hypervariability, called complementarity determining regions (CDRs), interspersed with regions that are more conserved, called framework regions (FRs). H and V L is composed of three CDRs and four FRs arranged from amino terminus to carboxy terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. The variable regions of the heavy and light chains contain a binding domain that interacts with an antigen. The constant region of the antibody may mediate the binding of the immunoglobulin to host tissues or factors, including various cells of the immune system (e.g., effector cells) and the first component (C1 q) of the classical complement system.

[0043] The term "human antibody," as used herein, is intended to include antibodies having variable regions in which both the framework and CDR regions are derived from human germline immunoglobulin sequences. Additionally, if the antibody contains a constant region, the constant region also is derived from a human germline immunoglobulin sequence. Human antibodies of the presently disclosed subject matter may include amino acid residues not encoded by human germline immunoglobulin sequences (e.g., mutations introduced by random or site-specific mutagenesis in vitro or by somatic mutation in vivo).

[0044] As used herein, the term "monoclonal antibody" refers to an antibody obtained from a population of substantially homogeneous antibodies, i.e., the individual antibodies constituting the population are identical and / or bind the same epitope, except for possible variant antibodies (such variants are generally present in minor amounts) that may include, for example, naturally occurring mutations or arise during production of the monoclonal antibody preparation. Furthermore, in contrast to polyclonal antibody preparations that typically include different antibodies that are directed against different determinants (epitopes), each monoclonal antibody of a monoclonal antibody preparation is directed against a single determinant on an antigen. Thus, the modifier "monoclonal" indicates the character of the antibody as obtained from a substantially homogeneous antibody population and should not be construed as requiring production of the antibody by any particular method. For example, monoclonal antibodies used in accordance with the subject matter disclosed herein may be produced by a variety of techniques, including, but not limited to, hybridoma methods, recombinant DNA methods, phage display methods, and methods utilizing transgenic animals that contain all or a portion of the human immunoglobulin loci, and such methods and other exemplary methods for producing monoclonal antibodies are described herein.

[0045] As used herein, the term "recombinant human antibody" includes all human antibodies prepared, expressed, generated, or isolated by recombinant means, such as (a) antibodies isolated from animals (e.g., mice) that are transgenic or transchromosomal for human immunoglobulin genes or hybridomas prepared therefrom (described further below), (b) antibodies isolated from host cells that have been transformed to express human antibodies, e.g., from transfectomas, (c) antibodies isolated from recombinant combinatorial human antibody libraries, and (d) antibodies prepared, expressed, generated, or isolated by any other means that involve splicing of human immunoglobulin gene sequences to other DNA sequences. Such recombinant human antibodies have variable regions in which the framework and CDR regions are derived from human germline immunoglobulin sequences. However, in certain embodiments, such recombinant human antibodies may be subjected to in vitro mutagenesis (or, when animals transgenic for human Ig sequences are used, in vivo somatic mutagenesis), thus allowing the V of the recombinant antibody to be modified. H Area and V L The amino acid sequence of the region is H Sequence and V L These are sequences that, while derived from and related to the sequence, may not naturally occur in the human antibody germline repertoire in vivo.

[0046] The term "humanized antibody" is intended to refer to antibodies in which CDR sequences derived from the germline of another mammalian species, such as a mouse, have been grafted onto human framework sequences. Additional framework region modifications may be made within the human framework sequences.

[0047] The term "chimeric antibody" is intended to refer to an antibody in which the variable region sequences are derived from one species and the constant region sequences are derived from another species, e.g., the variable region sequences are derived from a murine antibody and the constant region sequences are derived from a human antibody.

[0048] As used herein, an antibody that "specifically binds to uPAR" has a concentration of about 5×10 -7M or less, approximately 1×10 -7 M or less, about 5 x 10 -8 M or less, approximately 1×10 -8 M or less, about 5 x 10 -9 M or less, approximately 1×10 -9 M or less, about 5 x 10 -10 M or less, approximately 1×10 -10 M or less, about 5 x 10 -11 M or less, or about 1 x 10 -11 The dissociation constant (K D ) is intended to refer to an antibody that binds to uPAR (e.g., human uPAR).

[0049] An "antibody that competes for binding" or "antibody that cross-competes for binding" with a reference antibody for binding to an antigen, e.g., uPAR, refers to an antibody that blocks the binding of the reference antibody to an antigen (e.g., uPAR) by 50% or more in a competitive assay, and conversely, the reference antibody blocks the binding of the antibody to an antigen (e.g., uPAR) by 50% or more in a competitive assay. Exemplary competitive assays are described in "Antibodies", Harlow and Lane (Cold Spring Harbor Press, Cold Spring Harbor, NY).

[0050] As used herein, "isotype" refers to the antibody class (e.g., IgM or IgG1) that is encoded by heavy chain constant region genes.

[0051] The phrases "an antibody that recognizes an antigen" and "an antibody specific for an antigen" may be used interchangeably herein with the term "an antibody that specifically binds to an antigen (eg, a uPAR polypeptide)."

[0052] As used herein, the term "antigen-binding fragment" or "antigen-binding region" of an antibody refers to that region or fragment of an antibody that binds to an antigen and confers antigen specificity to the antibody, and includes fragments of antigen-binding proteins, e.g., antibodies, including one or more fragments of an antibody that retain the ability to specifically bind to an antigen (e.g., a uPAR polypeptide). It has been shown that the antigen-binding function of an antibody can be performed by fragments of a full-length antibody. Examples of antigen-binding fragments encompassed by the term "antibody fragment" of an antibody include V L Domain, V H Domain, C L a Fab fragment, which is a monovalent fragment consisting of a CH1 domain and a CH2 domain; a F(ab)2 fragment, which is a bivalent fragment containing two Fab fragments linked by a disulfide bridge at the hinge region; H an Fd fragment consisting of the V domain and the CH1 domain of a single arm of an antibody; L Domain and V H Fv fragment consisting of domains, V H These include dAb fragments consisting of domains (Ward et al., Nature 1989;341:544-546), as well as isolated complementarity determining regions (CDRs).

[0053] In addition, the two domains V of the Fv fragment L and V H are encoded by separate genes, but these are referred to as V L Area and V H The antibody fragments can be joined using recombinant methods by synthetic linkers that allow the domains to be produced as a single protein chain that pairs to form a monovalent molecule. These are known as single-chain Fvs (scFvs), see, e.g., Bird et al., Science (1988); 242:423-426; and Huston et al., Proc Natl Acad Sci (1998); 85:5879-5883. These antibody fragments are obtained using conventional techniques known to those of skill in the art, and the fragments are screened for utility in the same manner as intact antibodies.

[0054] An "antibody" or "antigen binding protein" is one that has been identified, separated and / or recovered from a component of its natural environment. A "synthetic antibody" or "recombinant antibody" is generally produced using recombinant techniques, or using peptide synthesis techniques known to those of skill in the art.

[0055] As used herein, the term "single chain variable fragment" or "scFv" refers to a V H ::V L Heavy chains (V) of immunoglobulins (e.g., mouse or human) covalently linked to form heterodimers H ) and light chain (V L ) is a fusion protein of the variable regions of the heavy chain (V H ) and light chain (V L ) are either directly connected or connected by a peptide-encoding linker (e.g., 10, 15, 20, 25 amino acids), H N-terminus of V L or V H The C-terminus of V L The linker is usually rich in glycine for flexibility and rich in serine or threonine for solubility. The linker may link the heavy and light chain variable regions of the extracellular antigen-binding domain.

[0056] Non-limiting examples of linkers are described in Shen et al., Anal Chem (2008); 80(6): 1910-1917 and WO2014 / 087010, the contents of which are incorporated herein by reference in their entirety. In certain embodiments, the linker is a G4S linker. In certain embodiments, the linker comprises or consists of the amino acid sequence set forth in SEQ ID NO: 62, provided below. GGGGSGGGGSGGGSGGGGS [SEQ ID NO: 62]

[0057] In certain embodiments, the linker comprises or consists of the amino acid sequence set forth in SEQ ID NO:63, provided below. GGGGSGGGGSGGGGS [SEQ ID NO: 63]

[0058] In certain embodiments, the linker comprises or consists of the amino acid sequence set forth in SEQ ID NO:64, provided below. GGGGSGGGGSGGGGSGGGSGGGGS [SEQ ID NO: 64]

[0059] In certain embodiments, the linker comprises or consists of the amino acid sequence set forth in SEQ ID NO:65, provided below. GGGGSGGGGSGGGGSGGGGSGGGSGGGGS [SEQ ID NO: 65]

[0060] In certain embodiments, the linker comprises or consists of the amino acid sequence set forth in SEQ ID NO:66, provided below. GGGGS [SEQ ID NO: 66]

[0061] In certain embodiments, the linker comprises or consists of the amino acid sequence set forth in SEQ ID NO:67, provided below. GGGGSGGGGS [SEQ ID NO: 67]

[0062] Despite the removal of the constant region and the introduction of the linker, the scFv protein retains the specificity of the original immunoglobulin. Single-chain Fv polypeptide antibodies are synthesized by the VFv method, as described by Huston, et al. (Proc. Nat. Acad. Sci. USA, 1988; 85:5879-5883). H and V LThe polypeptide can be expressed from a nucleic acid containing a sequence encoding the polypeptide. See also U.S. Patent Nos. 5,091,513, 5,132,405, and 4,956,778, and U.S. Patent Publication Nos. 2005 / 0196754 and 2005 / 0196754. Antagonistic scFvs with inhibitory activity have been described (e.g., Zhao et al., Hyrbidoma(Larchmt) 2008;27(6):455-51; Peter et al., J Cachexia Sarcopenia Muscle 2012 August 12; Shieh et al., J Imunol 2009;183(4):2277-85; Giomarelli et al., Thromb Haemost 2007;97(6):955-63; Fife et al., J Clin Invst 2006;116(8):2252-61; Brocks et al., Immunotechnology 1997;3(3):173-84; Moosmayer et al., Ther Immunol 1995;2 (10:31-40). Agonistic scFvs with stimulatory activity have been described (see, e.g., Peter et al., J Bioi Chern 2003;25278(38):36740-7; Xie et al., Nat Biotech 1997;15(8):768-71; Ledbetter et al., Crit Rev Immunol 1997;17(5-6):427-55; Ho et al., BioChim Biophys Acta 2003;1638(3):257-66).

[0063] As used herein, "F(ab)" refers to the fragment of an antibody structure that binds an antigen but is monovalent and does not have the Fc portion; for example, an antibody digested with the enzyme papain produces two F(ab) fragments and an Fc fragment (the heavy (H) chain constant region, the Fc region that does not bind antigen).

[0064] As used herein, "F(ab')2" refers to an antibody fragment produced by pepsin digestion of a whole IgG antibody, which has two antigen-binding (ab') (bivalent) regions, each (ab') region containing two separate amino acid chains of a part of a H chain and a light (L) chain linked by S-S bonds to bind the antigen, and the remaining parts of the H chain are linked together. The "F(ab')2" fragment can be divided into two individual Fab' fragments.

[0065] As used herein, the term "vector" refers to any genetic element, such as a plasmid, phage, transposon, cosmid, chromosome, virus, virion, etc., that is capable of replicating and transferring genetic sequences into a cell when associated with the appropriate control elements. Thus, the term includes cloning and expression vehicles, as well as viral vectors and plasmid vectors.

[0066] "CDR" is defined as the complementarity determining region amino acid sequences of an antibody that are hypervariable regions of immunoglobulin heavy and light chains. See, e.g., Kabat et al., Sequences of Proteins of Immunological Interest, 4th USDepartment of Health and Human Services, National Institutes of Health (1987), Chothia, or IMGT numbering system (Lefranc, The Immunologist (1999); 7:132-136; Lefranc et al., Dev. Comp. Immunol. (2003); 27:55-77). In certain embodiments, CDRs are identified using the IMGT numbering system, accessible at http: / / www.imgt.org / IMGT_vquest / input. The term "hypervariable region" or "HVR" as used herein refers to each of the regions of an antibody variable domain that are hypervariable in sequence ("complementarity determining region" or "CDR") and / or form structurally defined loops ("hypervariable loops") and / or contain antigen contact residues ("antigen contacts"). Generally, an antibody contains three heavy chain and three light chain CDRs or CDR regions in the variable region. The CDRs provide the majority of contact residues for the binding of the antibody to the antigen or epitope region. In certain embodiments, the CDRs are identified according to the Kabat numbering system. In certain embodiments, the CDRs are identified according to the Kabat numbering system and the Chothia system.

[0067] The term "isolated" denotes the degree of separation from the original source or surroundings.

[0068] An "isolated antibody" is one that has been separated from a component of its natural environment. In certain embodiments, the antibody is purified to greater than 95% or 99% purity, for example, as determined by electrophoresis (e.g., SDS-PAGE, isoelectric focusing (IEF), capillary electrophoresis) or chromatography (e.g., ion exchange or reverse phase HPLC). For a review of methods for assessing antibody purity, see, for example, Flatman et al., J. Chromatogr (2007); B 848: 79-87.

[0069] "Isolated nucleic acid" refers to a nucleic acid molecule that has been separated from a component of its natural environment. Isolated nucleic acid includes a nucleic acid molecule contained within a cell that ordinarily contains the nucleic acid molecule, but where the nucleic acid molecule is present extrachromosomally or at a chromosomal location that is different from its natural chromosomal location.

[0070] "Isolated nucleic acid encoding an antibody" (including reference to a specific antibody, e.g., an anti-KLB antibody) refers to one or more nucleic acid molecules encoding antibody heavy and light chains (or fragments thereof), including such nucleic acid molecules within a single vector or separate vectors, and such nucleic acid molecules are present in one or more locations within a host cell.

[0071] The term "vector", as used herein, is intended to refer to a nucleic acid molecule capable of propagating another nucleic acid to which it is linked. The term includes vectors as self-replicating nucleic acid structures and vectors that integrate into the genome of a host cell into which they are introduced. Certain vectors are capable of directing the expression of genes to which they are operatively linked. Such vectors are referred to herein as "expression vectors".

[0072] An "immunoconjugate" is an antibody conjugated to one or more heterologous molecules, including, but not limited to, cytotoxic agents.

[0073] An "effective amount" (or "therapeutically effective amount") is an amount sufficient to produce a beneficial or desired clinical result upon treatment. An effective amount may be administered to a subject in one or more doses. In terms of treatment, an effective amount is an amount sufficient to palliate, ameliorate, stabilize, reverse or slow the progression of a disease, or otherwise reduce the pathological consequences of a disease. An effective amount is generally determined by a physician on a case-by-case basis and is within the skill of one of ordinary skill in the art. Several factors are typically considered in determining the appropriate dosage to achieve an effective amount. These factors include the age, sex, and weight of the subject, the condition being treated, the severity of the condition, and the form and effective concentration of the cells to be administered.

[0074] An "individual" or "subject" herein is a vertebrate, e.g., a human or a non-human animal, e.g., a mammal. Mammals include, but are not limited to, humans, primates, farm animals, sport animals, rodents, and pets. Non-limiting examples of non-human animal subjects include rodents, such as mice, rats, and hamsters, guinea pigs, rabbits, dogs, cats, sheep, pigs, goats, cows, horses, and non-human primates, such as apes and monkeys.

[0075] As used herein, "treatment" (and grammatical variations such as "treat" or "treating") refers to clinical intervention in an attempt to alter the natural course of the individual being treated, and can be performed either for prophylaxis or during the course of clinical pathology. Desirable effects of treatment include, but are not limited to, preventing the onset or recurrence of disease, alleviation of symptoms, alleviation of any direct or indirect pathological consequences of the disease, preventing metastasis, reducing the rate of disease progression, amelioration or amelioration of the disease state, and remission or improved prognosis. In certain embodiments, the antibodies of the subject matter disclosed herein are used to delay the onset of disease or to slow the progression of a disease, e.g., a tumor, e.g., a tumor associated with uPAR.

[0076] The terms "comprises" and "comprising" are intended to have the broad meaning ascribed to them in U.S. patent law and can mean "includes," "including," etc.

[0077] As used herein, the term "about" or "approximately" means within an acceptable error range for a particular value as determined by one of ordinary skill in the art, which depends in part on how the value is measured or determined, i.e., the limitations of the measurement system. For example, "about" can mean within 3 or more than 3 standard deviations, as is customary in the art. Alternatively, "about" can mean within a range of up to 20%, preferably up to 10%, more preferably up to 5%, and even more preferably up to 1% of a given value. Alternatively, particularly with respect to biological systems or processes, the term can mean within an order of magnitude, preferably within 5-fold, and more preferably within 2-fold of a value.

[0078] As described herein, any concentration range, percentage range, ratio range, or integer range, unless otherwise indicated, should be understood to include any integer value within the recited range, and fractions thereof, where appropriate (such as tenths and hundredths of integers).

[0079] Other aspects of the presently disclosed subject matter are described in the disclosure that follows and are within the scope of the presently disclosed subject matter.

[0080] 4.2.uPAR uPAR (urokinase-type plasminogen activator receptor), also known as CD87, is a glycosylphosphatidylinositol-anchored protein. uPAR is rich in cysteines and consists of three tandem LU domains that bind urokinase-type plasminogen activator (uPA). (Kessler et al., J.Neurochem.(2017);142:7-18; Llinas et al., EMBOJ.(2005);24(9):1655-63; Huai et al., Science(2006);311(5761):656-9; Chelsea et al., Human Genomics(2016);10:10). uPAR also interacts with several other proteins, including vitronectin, uPAR-associated protein (uPARAP), and the integrin family of membrane proteins.

[0081] uPAR has been implicated in tumor growth or metastasis in a variety of different types of cancer, including breast cancer (including triple-negative breast cancer), endometrial cancer, ovarian cancer, colon cancer, rectal cancer, lung cancer, stomach cancer, prostate cancer, renal cancer, pancreatic cancer, rectal cancer, cervical cancer, head and neck cancer, liver cancer, gastric cancer, urothelial cancer, melanoma, brain cancer (including glioblastoma multiforme), acute lymphoblastic leukemia (ALL), chronic lymphocytic leukemia (CLL), and acute myeloid leukemia (AML). It also plays a role in aging, including its association with aging-related diseases associated with aging. It can also regulate immune responses and cell-matrix interactions, promoting tumor cell proliferation and emergence from dormancy.

[0082] uPAR is induced during the process of cellular senescence, can be induced by certain cancer drugs, and can accumulate in a range of age-related and tissue-damaging pathologies (LIST). Removal of senescent cells can improve therapy responses and alleviate symptoms of tissue-damaging pathologies, including fibrosis.

[0083] Soluble urokinase plasminogen activator receptor (suPAR) is found to be upregulated in some of the above pathologies, as well as in chronic obstructive pulmonary disease, asthma, liver failure, heart failure, cardiovascular disease, and rheumatoid arthritis. (Desmedt et al., Crit. Rev. Clin. Lab. Sci. (2017); 54(2): 117-133). uPAR is found to be highly expressed on senescent cells. (Wagner et al., Nature (2020); 583 (7814): 37-38, Amor et al., Nature (2020 Jul); 583 (7814): 127-132). Thus, uPAR (e.g., suPAR) can be used as a biomarker of disease stage. Many oncogenic signaling pathways and tumor microenvironment conditions, such as hypoxia, can activate transcription factors and then regulate uPAR. uPAR can regulate protein degradation by associating with the outer layer of the plasma membrane via a glycosylphosphatidylinositol (GPI) anchor, but can be secreted or shed from the cell surface. (Harvey et al., Nat. Rev. Mol. Cell Biol (2010); 11, 23-36). uPAR expression directly correlates with the invasive potential of endometrial cancer. (Foca et al., Gynecol. Oncol. (2000); 79(2): 244-50). uPAR is involved in several hematological malignancies, particularly acute leukemia and multiple myeloma. (Hata et al., Blood (1993); 81: 3357-3364, MC Bene et al., Leukemia (2004); 18, 394-400). uPAR has been reported to be associated with poor prognosis in breast cancer patients. (Bo et al., Oncol.Rep.(2005);14(1):105-12, Foekens et al., Cancer Res.(2000);60(3):636-43).

[0084] In certain embodiments, uPAR is human uPAR comprising or consisting of the amino acid sequence of UniProt reference number Q03405-1 (SEQ ID NO: 61) or a fragment thereof. SEQ ID NO: 61 is provided below. In certain embodiments, uPAR comprises three domains: domain 1 (domain UPAR / Ly6 1), domain 2 (domain UPAR / Ly6 2), and domain 3 (domain UPAR / Ly6 3). In certain embodiments, domain 1 comprises or consists of amino acids 23-114 of SEQ ID NO: 61. In certain embodiments, domain 2 comprises or consists of amino acids 115-213 of SEQ ID NO: 61. In certain embodiments, domain 3 comprises or consists of amino acids 214-305 of SEQ ID NO: 61. [ka]

[0085] In certain embodiments, uPAR comprises or consists of an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99%, at least about 100% identical to the amino acid sequence set forth in SEQ ID NO: 61 or a fragment thereof.

[0086] In certain embodiments, the anti-uPAR antibody or antigen-binding fragment thereof binds to a portion of human uPAR. In certain embodiments, the anti-uPAR antibody or antigen-binding fragment thereof binds to at least one of domain 1, domain 2, and domain 3. In certain embodiments, the anti-uPAR antibody or antigen-binding fragment thereof binds to domain 2. In certain embodiments, the anti-uPAR antibody or antigen-binding fragment thereof binds to domain 3. In certain embodiments, the anti-uPAR antibody or antigen-binding fragment thereof binds to both domain 2 and domain 3. In certain embodiments, the anti-uPAR antibody or antigen-binding fragment thereof binds to amino acids 115-303 of SEQ ID NO:61. In certain embodiments, the anti-uPAR antibody or antigen-binding fragment thereof binds to amino acids 115-305 of SEQ ID NO:61.

[0087] 4.3. Anti-uPAR antibodies The antibodies of the presently disclosed subject matter are characterized by a particular functional feature or property of the antibody, for example, the antibody specifically binds to uPAR (e.g., binds to human uPAR).

[0088] In certain embodiments, the antibodies or antigen-binding fragments disclosed herein have a binding affinity, e.g., of 1×10 -6 M or less, for example, about 1×10 -7 M or less, approximately 1×10 -8 M or less, approximately 1×10 -9 M or less, approximately 1×10 -10 M or less, or about 1 x 10 -11 The dissociation constant (K D ) and binds to uPAR (e.g., human uPAR). In certain embodiments, the antibodies or antigen-binding fragments disclosed herein bind to uPAR at about 1×10 -7 K below M D In certain embodiments, the antibodies or antigen-binding fragments disclosed herein bind to uPAR (e.g., human uPAR) at about 1×10 -9 M ~ approx. 1×10 -7 K of M DIn certain embodiments, the antibodies or antigen-binding fragments disclosed herein bind to uPAR (e.g., human uPAR) at about 1×10 -8 K of M D In certain embodiments, the antibody or antigen-binding fragment disclosed herein binds to uPAR (e.g., human uPAR) at about 1.9×10 -8 K of M D In certain embodiments, the antibodies or antigen-binding fragments disclosed herein bind to uPAR (e.g., human uPAR) at about 2×10 -8 K of M D In certain embodiments, the antibodies or antigen-binding fragments disclosed herein bind to uPAR (e.g., human uPAR) at about 3×10 -8 K of M D In certain embodiments, the antibody or antigen-binding fragment disclosed herein binds to uPAR (e.g., human uPAR) at about 3.5×10 -8 K of M D In certain embodiments, the antibodies or antigen-binding fragments disclosed herein bind to uPAR (e.g., human uPAR) at about 4×10 -8 K of M D In certain embodiments, the antibody or antigen-binding fragment disclosed herein binds to uPAR (e.g., human uPAR) at about 4.2×10 -8 K of M D In certain embodiments, the antibodies or antigen-binding fragments disclosed herein bind to uPAR (e.g., human uPAR) at about 1×10 -7 K of M D In certain embodiments, the antibody or antigen-binding fragment disclosed herein binds to uPAR (e.g., human uPAR) at about 9.5×10 -8 K of M D In certain embodiments, the antibodies or antigen-binding fragments disclosed herein bind to uPAR (e.g., human uPAR) at about 6×10 -9 K of M D In certain embodiments, the antibodies or antigen-binding fragments disclosed herein bind to uPAR (e.g., human uPAR) at about 7×10-9 K of M D In certain embodiments, the antibody or antigen-binding fragment disclosed herein binds to uPAR (e.g., human uPAR) at about 6.6×10 -9 K of M D and binds to uPAR (e.g., human uPAR).

[0089] The heavy and light chains of the antibodies or antigen-binding fragments disclosed herein can be full length (e.g., an antibody can comprise at least one (e.g., one or two) complete heavy chains and at least one (e.g., one or two) complete light chains) or can comprise an antigen-binding fragment (Fab, F(ab')2, Fv, or single chain Fv fragment ("scFv")). In certain embodiments, the antibody heavy chain constant region is selected from, e.g., IgG1, IgG2, IgG3, IgG4, IgM, IgA1, IgA2, IgD, and IgE, particularly selected from, e.g., IgG1, IgG2, IgG3, and IgG4. In certain embodiments, the immunoglobulin isotype is IgG1 (e.g., human IgG1). In certain embodiments, the antibody light chain constant region is selected from, e.g., kappa or lambda, particularly kappa.

[0090] 4.3.1. Single-chain variable fragments (scFv) In certain embodiments, the subject matter disclosed herein includes antibodies or antigen-binding fragments thereof in which the scFv sequence is fused to one or more constant domains to form an antibody with the Fc region of a human immunoglobulin to produce a bivalent protein and increase the overall affinity and stability of the antibody. In addition, the Fc portion allows other molecules to be directly conjugated to the antibody, including but not limited to fluorescent dyes, cytotoxins, radioisotopes, etc., for use in, for example, antigen quantification studies, to immobilize the antibody for affinity measurements, for targeted delivery of therapeutic agents, to test Fc-mediated cytotoxicity using immune effector cells, and many other applications.

[0091] The results presented herein highlight the specificity, sensitivity and utility of the antibodies or antigen-binding fragments disclosed herein in targeting uPAR polypeptides (eg, human uPAR).

[0092] The molecules disclosed herein are based on the identification and selection of single chain variable fragments (scFv) using phage display, the amino acid sequence of which confers the specificity of the molecule to the uPAR polypeptide of interest and forms the basis of all antigen binding proteins of the present disclosure. Thus, scFv can be used to design a diverse family of "antibody" molecules, including, for example, full-length antibodies, fragments thereof such as Fab and F(ab')2, minibodies, fusion proteins including scFv-Fc fusions, multivalent antibodies, i.e., antibodies with multiple specificities against the same or different antigens, e.g., multispecific antibodies, tribodies, etc. (Cuesta et al., Multivalent antibodies: when design surpasses evolution. Trends in Biotechnology 28:355-362 2010).

[0093] In certain embodiments, the antigen-binding protein is a full-length antibody, and the heavy and light chains of the antibody of the presently disclosed subject matter can be full-length (e.g., the antibody can include at least one, or two complete heavy chains, and at least one, preferably two complete light chains) or can include an antigen-binding fragment (Fab, F(ab')2, Fv, or scFv). In certain embodiments, the antibody heavy chain constant region is selected from IgG1, IgG2, IgG3, IgG4, IgM, IgA1, IgA2, IgD, and IgE, etc. In certain embodiments, the immunoglobulin isotype is selected from IgG1, IgG2, IgG3, and IgG4. In certain embodiments, the immunoglobulin isotype is IgG1 (e.g., human IgG1). The choice of antibody isotype can depend on the immune effector function that the antibody is designed to elicit.

[0094] In constructing recombinant immunoglobulins, the appropriate amino acid sequences for the constant regions of various immunoglobulin isotypes and methods for producing a broad range of antibodies are known to those of skill in the art.

[0095] In certain embodiments, the anti-uPAR scFv comprises the amino acid sequence set forth in SEQ ID NO:25. H , V comprising the amino acid sequence set forth in SEQ ID NO:26 L and optionally a linker sequence, e.g., a linker peptide, between the heavy chain variable region and the light chain variable region. SEQ ID NOs: 25 and 26 are provided in Table 1 below. In certain embodiments, the scFv is designated "8B1".

[0096] In certain embodiments, the anti-uPAR scFv is a V selected from Table 1. H Area and V L In certain embodiments, the anti-uPAR scFv is a scFv-Fc fusion protein or a full-length human IgG having a V domain or CDR. H In certain embodiments, the anti-uPAR scFv comprises a V L In certain embodiments, the anti-uPAR scFv comprises a V H and V comprising the amino acid sequence set forth in SEQ ID NO:26 L Including,

[0097] In certain embodiments, the anti-uPAR scFv comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO:1 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO:2 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO:3 or a conservative modification thereof. H SEQ ID NOs: 1-3 are provided in Table 1.

[0098] In certain embodiments, the anti-uPAR scFv comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO:4 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO:5 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO:6 or a conservative modification thereof. L SEQ ID NOs: 4-6 are provided in Table 1.

[0099] In certain embodiments, the anti-uPAR scFv comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO:1 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO:2 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO:3 or a conservative modification thereof. H and CDR1 comprising the amino acid sequence set forth in SEQ ID NO:4 or a conservative modification thereof, CDR2 comprising the amino acid sequence set forth in SEQ ID NO:5 or a conservative modification thereof, and CDR3 comprising the amino acid sequence set forth in SEQ ID NO:6 or a conservative modification thereof. L Including,

[0100] In certain embodiments, the anti-uPAR scFv comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO:1, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO:2, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO:3. H and CDR1 comprising the amino acid sequence set forth in SEQ ID NO:4, CDR2 comprising the amino acid sequence set forth in SEQ ID NO:5, and CDR3 comprising the amino acid sequence set forth in SEQ ID NO:6.

[0101] In certain embodiments, the anti-uPAR scFv comprises the amino acid sequence set forth in SEQ ID NO:25. H and V comprising the amino acid sequence set forth in SEQ ID NO:26 L In certain embodiments, V H and V L are linked via a linker. An exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO:25 is set forth in SEQ ID NO:33. An exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO:26 is set forth in SEQ ID NO:34. SEQ ID NOs:33 and 34 are provided in Table 1 below.

[0102] In certain embodiments, the variable region is a heavy chain variable region (V H ) are located at the N-terminus. In certain embodiments, the variable regions are arranged from the N-terminus to the C-terminus: H -V L In certain embodiments, the light chain variable region (V L ) is positioned at the N-terminus. In certain embodiments, the variable regions are positioned from N-terminus to C-terminus: L -V H .

[0103] In certain embodiments, the CDR sequences disclosed in Table 1 are identified according to the Kabat system or a combination of the Kabat and Chothia systems. SEQ ID NOs: 2-6 are identified according to the Kabat system. SEQ ID NO: 1 is identified according to a combination of the Kabat and Chothia systems. [Table 1]

[0104] In certain embodiments, the anti-uPAR scFv comprises the amino acid sequence set forth in SEQ ID NO:27. H , V comprising the amino acid sequence set forth in SEQ ID NO:28 L and optionally a linker sequence, e.g., a linker peptide, between the heavy chain variable region and the light chain variable region. SEQ ID NOs: 27 and 28 are provided in Table 2 below. In certain embodiments, the scFv is designated "11E10".

[0105] In certain embodiments, the anti-uPAR scFv is a V selected from Table 2. H Area and V L In certain embodiments, the anti-uPAR scFv is a scFv-Fc fusion protein or a full-length human IgG having a V domain or CDR. HIn certain embodiments, the anti-uPAR scFv comprises a V L In certain embodiments, the anti-uPAR scFv comprises a V H and V comprising the amino acid sequence set forth in SEQ ID NO:28 L Including,

[0106] In certain embodiments, the anti-uPAR scFv comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO:7 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO:8 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO:9 or a conservative modification thereof. H SEQ ID NOs: 7-9 are provided in Table 2.

[0107] In certain embodiments, the anti-uPAR scFv comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 10 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 11 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 12 or a conservative modification thereof. L SEQ ID NOs: 10 to 12 are provided in Table 2.

[0108] In certain embodiments, the anti-uPAR scFv comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO:7 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO:8 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO:9 or a conservative modification thereof. H and CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 10 or a conservative modification thereof, CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 11 or a conservative modification thereof, and CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 12 or a conservative modification thereof. L Including,

[0109] In certain embodiments, the anti-uPAR scFv comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO:7, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO:8, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO:9. H and CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 10, CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 11, and CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 12. L Includes.

[0110] In certain embodiments, the anti-uPAR scFv comprises the amino acid sequence set forth in SEQ ID NO:27. H and V comprising the amino acid sequence set forth in SEQ ID NO:28 L In certain embodiments, V H and V L are linked via a linker. An exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO:27 is set forth in SEQ ID NO:35. An exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO:28 is set forth in SEQ ID NO:36. SEQ ID NOs:35 and 36 are provided in Table 2 below.

[0111] In certain embodiments, the heavy chain variable region (V H ) is positioned at the N-terminus. In certain embodiments, the variable region is positioned from N-terminus to C-terminus: H -V L In certain embodiments, the light chain variable region (V L ) is positioned at the N-terminus. In certain embodiments, the variable region is positioned from N-terminus to C-terminus: L -V H .

[0112] In certain embodiments, the CDR sequences disclosed in Table 2 are identified according to the Kabat system or a combination of the Kabat and Chothia systems. SEQ ID NOs: 8-12 are identified according to the Kabat system. SEQ ID NO: 7 is identified according to a combination of the Kabat and Chothia systems. [Table 2-1] [Table 2-2]

[0113] In certain embodiments, the anti-uPAR scFv comprises the amino acid sequence set forth in SEQ ID NO:29. H , V comprising the amino acid sequence set forth in SEQ ID NO:30 H and optionally a linker sequence, e.g., a linker peptide, between the heavy chain variable region and the light chain variable region. SEQ ID NOs: 29 and 30 are provided in Table 3 below. In certain embodiments, the anti-uPAR scFv is referred to as "17C9".

[0114] In certain embodiments, the anti-uPAR scFv is a V selected from Table 3. H Area and V L In certain embodiments, the anti-uPAR scFv is a scFv-Fc fusion protein or a full-length human IgG having a V domain or CDR. H In certain embodiments, the anti-uPAR scFv comprises a V L Includes.

[0115] In certain embodiments, the anti-uPAR scFv comprises the amino acid sequence set forth in SEQ ID NO:29. H and V comprising the amino acid sequence set forth in SEQ ID NO:30 L Including,

[0116] In certain embodiments, the anti-uPAR scFv comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 13 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 14 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 15 or a conservative modification thereof. H SEQ ID NOs: 13-15 are provided in Table 3.

[0117] In certain embodiments, the anti-uPAR scFv comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 16 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 17 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 18 or a conservative modification thereof. L SEQ ID NOs: 16-18 are provided in Table 3.

[0118] In certain embodiments, the anti-uPAR scFv comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 13 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 14 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 15 or a conservative modification thereof. H and CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 16 or a conservative modification thereof, CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 17 or a conservative modification thereof, and CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 18 or a conservative modification thereof. L Including,

[0119] In certain embodiments, the anti-uPAR scFv comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 13, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 14, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 15. H and CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 16, CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 17, and CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 18. L Includes.

[0120] In certain embodiments, the anti-uPAR scFv comprises the amino acid sequence set forth in SEQ ID NO:29. H and V comprising the amino acid sequence set forth in SEQ ID NO:30 L In certain embodiments, V H and V Lare linked via a linker. An exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO:29 is set forth in SEQ ID NO:37. An exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO:30 is set forth in SEQ ID NO:38. SEQ ID NOs:37 and 38 are provided in Table 3 below.

[0121] In certain embodiments, the heavy chain variable region (V H ) is positioned at the N-terminus. In certain embodiments, the variable regions are positioned from N-terminus to C-terminus: H -V L In certain embodiments, the light chain variable region (V L ) is positioned at the N-terminus. In certain embodiments, the variable regions are positioned from N-terminus to C-terminus: L -V H .

[0122] In certain embodiments, the CDR sequences disclosed in Table 3 are identified according to the Kabat system or a combination of the Kabat and Chothia systems. SEQ ID NOs: 14-18 are identified according to the Kabat system. SEQ ID NO: 13 is identified according to a combination of the Kabat and Chothia systems. [Table 3]

[0123] In certain embodiments, the anti-uPAR scFv comprises the amino acid sequence set forth in SEQ ID NO:31. H , V comprising the amino acid sequence set forth in SEQ ID NO:32 L and optionally a linker sequence, e.g., a linker peptide, between the heavy chain variable region and the light chain variable region. SEQ ID NOs: 31 and 32 are provided in Table 4 below. In certain embodiments, the anti-uPAR scFv is designated "19D7".

[0124] In certain embodiments, the anti-uPAR scFv is a V selected from Table 4. H Area and V LIn certain embodiments, the anti-uPAR scFv is a scFv-Fc fusion protein or a full-length human IgG having a V domain or CDR. H In certain embodiments, the anti-uPAR scFv comprises a V L In certain embodiments, the anti-uPAR scFv comprises a V H and V comprising the amino acid sequence set forth in SEQ ID NO:32 L and SEQ ID NOs:31 and 32 are provided in Table 4.

[0125] In certain embodiments, the anti-uPAR scFv comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 19 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 20 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 21 or a conservative modification thereof. H SEQ ID NOs: 19-21 are provided in Table 4.

[0126] In certain embodiments, the anti-uPAR scFv comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 22 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 23 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 24 or a conservative modification thereof. L SEQ ID NOs:22-24 are provided in Table 4.

[0127] In certain embodiments, the anti-uPAR scFv comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 19 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 20 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 21 or a conservative modification thereof. H and CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 22 or a conservative modification thereof, CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 23 or a conservative modification thereof, and CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 24 or a conservative modification thereof. L Including,

[0128] In certain embodiments, the anti-uPAR scFv comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 19, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 20, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 21. H and CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 22, CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 23, and CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 24. L Includes.

[0129] In certain embodiments, the anti-uPAR scFv comprises the amino acid sequence set forth in SEQ ID NO:31. H and V comprising the amino acid sequence set forth in SEQ ID NO:32 L In certain embodiments, V H and V L are linked via a linker. An exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO:31 is set forth in SEQ ID NO:39. An exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO:32 is set forth in SEQ ID NO:40. SEQ ID NOs:39 and 40 are provided in Table 4 below.

[0130] In certain embodiments, the heavy chain variable region (V H ) is positioned at the N-terminus. In certain embodiments, the variable region is positioned from N-terminus to C-terminus: H -V L In certain embodiments, the light chain variable region (V L ) is positioned at the N-terminus. In certain embodiments, the variable region is positioned from N-terminus to C-terminus: L -V H .

[0131] In certain embodiments, the CDR sequences disclosed in Table 4 are identified according to the Kabat system or a combination of the Kabat and Chothia systems. SEQ ID NOs:20-24 are identified according to the Kabat system. SEQ ID NO:19 is identified according to a combination of the Kabat and Chothia systems. [Table 4]

[0132] In certain embodiments, the anti-uPAR scFv comprises the amino acid sequence set forth in SEQ ID NO:47. H , V comprising the amino acid sequence set forth in SEQ ID NO:48 L and optionally a linker sequence, e.g., a linker peptide, between the heavy chain variable region and the light chain variable region. SEQ ID NOs: 47 and 48 are provided in Table 5 below. In certain embodiments, the anti-uPAR scFv is designated "6C8".

[0133] In certain embodiments, the anti-uPAR scFv is a V selected from Table 5. H Area and V L In certain embodiments, the anti-uPAR scFv is a scFv-Fc fusion protein or a full-length human IgG having a V domain or CDR. H In certain embodiments, the anti-uPAR scFv comprises a V L In certain embodiments, the anti-uPAR scFv comprises a V H and V comprising the amino acid sequence set forth in SEQ ID NO:48 L and SEQ ID NOs: 47 and 48 are provided in Table 5.

[0134] In certain embodiments, the anti-uPAR scFv comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 41 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 42 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 43 or a conservative modification thereof. H SEQ ID NOs: 41-43 are provided in Table 5.

[0135] In certain embodiments, the anti-uPAR scFv comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 44 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 45 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46 or a conservative modification thereof. L SEQ ID NOs: 44-46 are provided in Table 5.

[0136] In certain embodiments, the anti-uPAR scFv comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 41 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 42 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 43 or a conservative modification thereof. H and CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 44 or a conservative modification thereof, CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 45 or a conservative modification thereof, and CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46 or a conservative modification thereof. L Including,

[0137] In certain embodiments, the anti-uPAR scFv comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 41, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 42, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 43. H and CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 44, CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 45, and CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46. L Includes.

[0138] In certain embodiments, the anti-uPAR scFv comprises the amino acid sequence set forth in SEQ ID NO:47. H and V comprising the amino acid sequence set forth in SEQ ID NO:48 L In certain embodiments, V H and V L are linked via a linker. An exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 47 is set forth in SEQ ID NO: 57. An exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 48 is set forth in SEQ ID NO: 58. SEQ ID NOs: 57 and 58 are provided in Table 5 below.

[0139] In certain embodiments, the heavy chain variable region (V H ) is positioned at the N-terminus. In certain embodiments, the variable region is positioned from N-terminus to C-terminus: H -V L In certain embodiments, the light chain variable region (V L ) is positioned at the N-terminus. In certain embodiments, the variable region is positioned from N-terminus to C-terminus: L -V H .

[0140] In certain embodiments, the CDR sequences disclosed in Table 5 are identified according to the Kabat system or a combination of the Kabat and Chothia systems. SEQ ID NOs: 42-46 are identified according to the Kabat system. SEQ ID NO: 41 is identified according to a combination of the Kabat and Chothia systems. [Table 5]

[0141] In certain embodiments, the anti-uPAR scFv comprises the amino acid sequence set forth in SEQ ID NO:59. H , V comprising the amino acid sequence set forth in SEQ ID NO:60 Land optionally a linker sequence, e.g., a linker peptide, between the heavy chain variable region and the light chain variable region. SEQ ID NOs: 59 and 60 are provided in Table 6 below. In certain embodiments, the anti-uPAR scFv is referred to as "14C5".

[0142] In certain embodiments, the anti-uPAR scFv is a V selected from Table 6. H Area and V L In certain embodiments, the anti-uPAR scFv is a scFv-Fc fusion protein or a full-length human IgG having a V domain or CDR. H In certain embodiments, the anti-uPAR scFv comprises a V L In certain embodiments, the anti-uPAR scFv comprises a V H and V comprising the amino acid sequence set forth in SEQ ID NO:60 L and SEQ ID NOs: 59 and 60 are provided in Table 6.

[0143] In certain embodiments, the anti-uPAR scFv comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 49 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 50 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 51 or a conservative modification thereof. H SEQ ID NOs: 49-51 are provided in Table 6.

[0144] In certain embodiments, the anti-uPAR scFv comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 52 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 53 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 54 or a conservative modification thereof. L SEQ ID NOs:52-54 are provided in Table 6.

[0145] In certain embodiments, the anti-uPAR scFv comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 49 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 50 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 51 or a conservative modification thereof. H , CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 52 or a conservative modification thereof, CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 53 or a conservative modification thereof, and CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 54 or a conservative modification thereof. L Includes.

[0146] In certain embodiments, the anti-uPAR scFv comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 49, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 50, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 51. H and CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 52, CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 53, and CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 54. L Includes.

[0147] In certain embodiments, the anti-uPAR scFv comprises the amino acid sequence set forth in SEQ ID NO:59. H and V comprising the amino acid sequence set forth in SEQ ID NO:60 L In certain embodiments, V H and V L are linked via a linker. An exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO:55 is set forth in SEQ ID NO:59. An exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO:56 is set forth in SEQ ID NO:60. SEQ ID NOs:59 and 60 are provided in Table 6 below.

[0148] In certain embodiments, the heavy chain variable region (V H ) is positioned at the N-terminus. In certain embodiments, the variable region is positioned from N-terminus to C-terminus: H -V L In certain embodiments, the light chain variable region (VL ) is positioned at the N-terminus. In certain embodiments, the variable regions are positioned from N-terminus to C-terminus: L -V H .

[0149] In certain embodiments, the CDR sequences disclosed in Table 6 are identified according to the Kabat system. [Table 6]

[0150] Monoclonal antibodies The presently disclosed subject matter provides antibodies (e.g., human antibodies, e.g., human monoclonal antibodies) that specifically bind to uPAR (e.g., human uPAR). H The amino acid sequences are set forth in SEQ ID NOs: 25, 27, 29, 31, 47, and 55, respectively. L The amino acid sequences are set forth in SEQ ID NOs: 26, 28, 30, 32, 48, and 56, respectively.

[0151] Given that each of the 8B1, 11E10, 17C9, 19D7, 6C8, and 14C5 antibodies can bind to uPAR, H The V and V sequences can be "mixed and matched" to create other anti-uPAR binding molecules. uPAR binding of such "mixed and matched" antibodies can be tested using binding assays known in the art, including, for example, ELISA, Western blot, RIA, Biacore analysis. Preferably, the V H and V L When chains are mixed and matched, a particular V H / V L V from involution H The sequence is structurally similar to V H Similarly, a particular V H / V LV from involution L The sequence is structurally similar to V L It is replaced by an array.

[0152] In certain embodiments, the subject matter disclosed herein comprises a heavy chain variable region (V) comprising an amino acid sequence selected from SEQ ID NOs: 25, 27, 29, 31, 47, and 55. H ) and (b) a light chain variable region (V L and V, wherein the antibody or antigen-binding fragment specifically binds to uPAR, e.g., human uPAR. H and V L teeth, (a) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 25, and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 26; or (b) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 27, and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 28; (c) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 29, and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 30; (d) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 31, and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 32; (e) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 47, and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 48; and (f) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO:55, and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:56.

[0153] In certain embodiments, the subject matter disclosed herein provides antibodies or antigen-binding fragments thereof that comprise the heavy and light chain CDR1, CDR2, and CDR3 of 8B1, 11E10, 17C9, 19D7, 6C8, and 14C5.

[0154] 8B1, 11E10, 17C9, 19D7, 6C8, and 14C5 V H The amino acid sequences of CDR1 are shown in SEQ ID NOs: 1, 7, 13, 19, 41, and 49, respectively. The V sequences of 8B1, 11E10, 17C9, 19D7, 6C8, and 14C5 antibodies are shown in SEQ ID NOs: 2, 8, 14, 20, 42, and 50, respectively. H Amino acid sequences of CDR2. V of 8B1, 11E10, 17C9, 19D7, 6C8, and 14C5 are set forth in SEQ ID NOs: 3, 9, 15, 21, 43, and 51, respectively. H Amino acid sequence of CDR3.

[0155] 8B1, 11E10, 17C9, 19D7, 6C8, and 14C5 V L The amino acid sequences of CDR1 are set forth in SEQ ID NOs: 4, 10, 16, 22, 44, and 52, respectively. L The amino acid sequences of CDR2 are set forth in SEQ ID NOs: 5, 11, 17, 23, 45, and 53, respectively. L The amino acid sequences of CDR3 are set forth, respectively, in SEQ ID NOs: 6, 12, 18, 24, 46, and 54. The CDR regions are delineated using the Kabat system, the Chothia system, or a combination thereof.

[0156] Given that each of these antibodies or antigen-binding fragments thereof can bind to uPAR, with antigen-binding specificity provided primarily by the CDR1, CDR2, and CDR3 regions, V H CDR1 sequence, V H CDR2 sequence and V H CDR3 sequence, and V L CDR1 sequence, V L CDR2 sequence and V L CDR3 sequences can be "mixed and matched" (i.e., CDRs from different antibodies can be mixed and matched, but each antibody can have the same V H CDR1, CDR2 and CDR3, and V L CDR1, VL CDR2 and V L Other anti-uPAR binding molecules can be made that contain the CDR3 of the antibody. The uPAR binding of such "mixed and matched" antibodies can be tested using the binding assays described above. H When CDR sequences are mixed and matched, a particular V H The CDR1, CDR2, and / or CDR3 sequences derived from the sequence are replaced with structurally similar CDR sequences. L When CDR sequences are mixed and matched, a particular V L The CDR1, CDR2 and / or CDR3 sequences derived from the sequence are preferably replaced with structurally similar CDR sequences. H Sequence and V L The sequence is a structurally similar sequence derived from the CDR sequences of the antibodies or antigen-binding fragments thereof disclosed herein (8B1, 11E10, 17C9, 19D7, 6C8, and 14C5), and includes one or more V H CDR region sequence and / or V L It will be readily apparent that such modifications can be made by substituting CDR region sequences.

[0157] In certain embodiments, the subject matter disclosed herein comprises: (a) a heavy chain variable region CDR1 comprising an amino acid sequence selected from SEQ ID NO:1, SEQ ID NO:7, SEQ ID NO:13, SEQ ID NO:19, SEQ ID NO:41, or SEQ ID NO:49; (b) a heavy chain variable region CDR2 comprising an amino acid sequence selected from SEQ ID NO: 2, SEQ ID NO: 8, SEQ ID NO: 14, SEQ ID NO: 20, SEQ ID NO: 42, or SEQ ID NO: 50; (c) a heavy chain variable region CDR3 comprising an amino acid sequence selected from SEQ ID NO:3, SEQ ID NO:9, SEQ ID NO:15, SEQ ID NO:21, SEQ ID NO:43, or SEQ ID NO:51; (d) a light chain variable region CDR1 comprising an amino acid sequence selected from SEQ ID NO: 4, SEQ ID NO: 10, SEQ ID NO: 16, SEQ ID NO: 22, SEQ ID NO: 44, or SEQ ID NO: 52; (e) a light chain variable region CDR2 comprising an amino acid sequence selected from SEQ ID NO:5, SEQ ID NO:11, SEQ ID NO:17, SEQ ID NO:23, SEQ ID NO:45, or SEQ ID NO:53; and (f) providing an antibody or antigen-binding fragment thereof, comprising a light chain variable region CDR3 comprising an amino acid sequence selected from SEQ ID NO:6, SEQ ID NO:12, SEQ ID NO:18, SEQ ID NO:24, SEQ ID NO:46, or SEQ ID NO:54.

[0158] In certain embodiments, the antibody or antigen-binding fragment thereof comprises: (a) a heavy chain variable region CDR1 comprising the amino acid sequence set forth in SEQ ID NO:1; (b) a heavy chain variable region CDR2 comprising the amino acid sequence set forth in SEQ ID NO:2; (c) a heavy chain variable region CDR3 comprising the amino acid sequence set forth in SEQ ID NO:3; (d) a light chain variable region CDR1 comprising the amino acid sequence set forth in SEQ ID NO:4; (e) a light chain variable region CDR2 comprising the amino acid sequence set forth in SEQ ID NO:5; and (f) comprising a light chain variable region CDR3 comprising the amino acid sequence set forth in SEQ ID NO:6.

[0159] In certain embodiments, the antibody or antigen-binding fragment comprises: (a) a heavy chain variable region CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 7; (b) a heavy chain variable region CDR2 comprising the amino acid sequence set forth in SEQ ID NO:8; (c) a heavy chain variable region CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 9; (d) a light chain variable region CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 10; (e) a light chain variable region CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 11; and (f) a light chain variable region CDR3 comprising the amino acid sequence set forth in SEQ ID NO:12.

[0160] In certain embodiments, the antibody or antigen-binding fragment thereof comprises: (a) a heavy chain variable region CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 13; (b) a heavy chain variable region CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 14; (c) a heavy chain variable region CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 15; (d) a light chain variable region CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 16; (e) a light chain variable region CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 17; and (f) a light chain variable region CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 18.

[0161] In certain embodiments, the antibody or antigen-binding fragment thereof comprises: (a) a heavy chain variable region CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 19; (b) a heavy chain variable region CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 20; (c) a heavy chain variable region CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 21; (d) a light chain variable region CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 22; (e) a light chain variable region CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 23; and (f) a light chain variable region CDR3 comprising the amino acid sequence set forth in SEQ ID NO:24.

[0162] In certain embodiments, the antibody or antigen-binding fragment thereof comprises: (a) a heavy chain variable region CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 41; (b) a heavy chain variable region CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 42; (c) a heavy chain variable region CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 43; (d) a light chain variable region CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 44; (e) a light chain variable region CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 45; and (f) a light chain variable region CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46.

[0163] In certain embodiments, the antibody or antigen-binding fragment thereof comprises: (a) a heavy chain variable region CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 49; (b) a heavy chain variable region CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 50; (c) a heavy chain variable region CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 51; (d) a light chain variable region CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 52; (e) a light chain variable region CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 53; and (f) comprises a light chain variable region CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 54. The constant / framework regions of the anti-uPAR antibodies disclosed herein can be modified, for example, by amino acid substitutions to alter the properties of the antibody (e.g., to increase or decrease one or more of antigen binding affinity, Fc receptor binding, antibody carbohydrate, e.g., glycosylation, fucosylation, etc., number of cysteine ​​residues, effector cell function, effector cell function, complement function, or introduction of conjugation sites).

[0164] In certain embodiments, the anti-uPAR antibody disclosed herein is a fully human antibody, such as any one of 8B1, 11E10, 17C9, 19D7, 6C8, and 14C5. Fully human mAbs cause severe side effects when administered to humans, including anaphylactic and hypersensitivity reactions.

[0165] The use of phage display libraries has allowed the selection of large antibody repertoires for unique and rare Abs against highly defined epitopes (for further details on phage display, see McCafferty et al., Phage antibodies: filamentous phage displaying antibody variable domains. Nature, 348:552-554). Thus, rapid identification of human Fab or single-chain Fv (scFv) fragments that are highly specific for tumor antigen-derived peptide-MHC complex molecules has become possible. In addition, by using Fab fragments to engineer full-length monoclonal antibodies (mAbs), it is possible to directly generate therapeutic human mAbs, avoiding the several months of time-consuming work that is usually required to develop therapeutic mAbs. The subject matter disclosed herein involves, for example, the development of fully human mAbs that recognize human uPAR polypeptides (e.g., polypeptides having the amino acid sequence set forth in SEQ ID NO: 61) for cancer therapy.

[0166] 4.3.3.Homologous antibodies In certain embodiments, the antibodies or antigen-binding fragments thereof disclosed herein comprise heavy and light chain variable regions comprising amino acid sequences that are homologous or identical to the amino acid sequences of the antibodies described herein (e.g., 8B1, 11E10, 17C9, 19D7, 6C8, and 14C5 antibodies), and the antibodies or antigen-binding fragments thereof retain the desired functional properties of the anti-uPAR antibodies or antigen-binding fragments thereof of the subject matter disclosed herein.

[0167] For example, the presently disclosed subject matter includes an antibody, or antigen-binding fragment thereof, comprising a heavy chain variable region and a light chain variable region, (a) the heavy chain variable region comprises an amino acid sequence that is at least about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% homologous or identical to the amino acid sequence set forth in SEQ ID NO:25, SEQ ID NO:27, SEQ ID NO:29, SEQ ID NO:31, SEQ ID NO:47, or SEQ ID NO:55; (b) an antibody or antigen-binding fragment thereof, wherein the light chain variable region comprises an amino acid sequence that is at least about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% homologous or identical to the amino acid sequence set forth in SEQ ID NO:26, SEQ ID NO:28, SEQ ID NO:30, SEQ ID NO:48, or SEQ ID NO:56.

[0168] In certain embodiments, V H Amino acid sequence and / or V L The amino acid sequence can be at least about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% homologous or identical to the sequences set forth above. H Area and V L V with high (i.e., 80% or more) homology or identity to the region H Area and V L Antibodies having regions can be obtained by mutagenesis (e.g., site-directed or PCR-mediated mutagenesis) followed by testing of the encoded modified antibody for retained function (i.e., binding affinity) using the binding assays described herein.

[0169] As used herein, the percentage of homology between two amino acid sequences corresponds to the percentage of identity between the two sequences. The percentage of identity or homology between two sequences is a function of the number of identical positions shared by the two sequences (i.e., % homology = number of identical positions / total number of positions x 100), taking into account the number of gaps and the length of each gap that need to be introduced for optimal alignment of the two sequences. Comparison of sequences and determination of the percentage of identity between two sequences can be accomplished using a mathematical algorithm, as described in the non-limiting examples below.

[0170] The percent homology or identity between two amino acid sequences can be determined using the algorithm of E. Meyers and W. Miller (Comput Appl Biosci (1988); 14:11-17) incorporated into the ALIGN program (version 2.0) using a PAM120 weight residue table, a gap length penalty of 12, and a gap penalty of 4. In addition, the percent homology between two amino acid sequences can be determined using the algorithm of Needleman and Wunsch (J Mol Biol (1970); 48:444-453) incorporated into the GAP program in the GCG software package (available at www.gcg.com) using either a Blossum62 matrix or a PAM250 matrix, and gap weights of 16, 14, 12, 10, 8, 6, or 4, and length weights of 1, 2, 3, 4, 5, or 6.

[0171] Additionally or alternatively, the subject protein sequences described herein can further be used as a "query sequence" to perform searches against public databases, for example to identify related sequences. Such searches can be performed using the XBLAST program (version 2.0) of Altschul et al., J Mol Biol (1990); 215:403-10. To obtain amino acid sequences homologous to the antibody molecules of the present invention, BLAST protein searches can be performed with the XBLAST program, score=50, wordlength=3. To obtain gapped alignments for comparison purposes, Gapped BLAST can be utilized as described in Altschul et al., Nucleic Acids Res (1997); 25(17):3389-3402. When utilizing BLAST, Gapped BLAST programs, the default parameters of the respective programs (e.g., XBLAST and NBLAST) can be used.

[0172] 4.3.4. Antibodies with Conservative Modifications In certain embodiments, the antibody or antigen-binding fragment thereof disclosed herein comprises a heavy chain variable region comprising a CDR1 sequence, a CDR2 sequence, and a CDR3 sequence, and a light chain variable region comprising a CDR1 sequence, a CDR2 sequence, and a CDR3 sequence, one or more of which comprises a specific amino acid sequence based on the preferred antibodies described herein (e.g., 8B1, 11E10, 17C9, 19D7, 6C8, and 14C5 antibodies) or a conservative modification thereof, and the antibody retains the desired functional properties of the anti-uPAR antibody or antigen-binding fragment thereof of the presently disclosed subject matter. The presently disclosed subject matter includes an antibody or antigen-binding fragment thereof comprising a heavy chain variable region comprising a CDR1 sequence, a CDR2 sequence, and a CDR3 sequence, and a light chain variable region comprising a CDR1 sequence, a CDR2 sequence, and a CDR3 sequence, (a) the heavy chain variable region CDR3 sequence comprises an amino acid sequence selected from the amino acid sequences of SEQ ID NOs: 3, 9, 15, 21, 43, and 51, and conservative modifications thereof; (b) an antibody or antigen-binding fragment thereof, wherein the light chain variable region CDR3 sequence comprises an amino acid sequence selected from the amino acid sequences of SEQ ID NOs: 6, 12, 18, 24, 46, and 54, and conservative modifications thereof.

[0173] In certain embodiments, the heavy chain variable region CDR3 sequence comprises an amino acid sequence selected from the amino acid sequences of SEQ ID NOs: 3, 9, 15, 21, 43, and 51, and conservative modifications thereof, and the light chain variable region CDR3 sequence comprises an amino acid sequence selected from the amino acid sequences of SEQ ID NOs: 6, 12, 18, 24, 46, and 54, and conservative modifications thereof.

[0174] In certain embodiments, the heavy chain variable region CDR2 sequence comprises an amino acid sequence selected from the amino acid sequences of SEQ ID NOs: 2, 8, 14, 20, 42, and 50, and conservative modifications thereof, and the light chain variable region CDR2 sequence comprises an amino acid sequence selected from the amino acid sequences of SEQ ID NOs: 5, 11, 17, 23, 45, and 53, and conservative modifications thereof.

[0175] In certain embodiments, the heavy chain variable region CDR1 sequence comprises an amino acid sequence selected from the amino acid sequences of SEQ ID NOs: 1, 7, 13, 19, 41, and 49, and conservative modifications thereof, and the light chain variable region CDR1 sequence comprises an amino acid sequence selected from the amino acid sequences of SEQ ID NOs: 4, 10, 16, 22, 44, and 52, and conservative modifications thereof.

[0176] As used herein, the term "conservative sequence modifications" is intended to refer to amino acid modifications that do not significantly affect or significantly alter the binding characteristics of the antibody containing the amino acid sequence. Such conservative modifications include amino acid substitutions, additions, and deletions. Modifications can be introduced into the antibodies of the disclosure by standard techniques known in the art, such as site-directed mutagenesis and PCR-mediated mutagenesis.

[0177] A conservative amino acid substitution is one in which an amino acid residue is replaced with an amino acid residue having a similar side chain. Families of amino acid residues having similar side chains have been defined in the art. Representative conservative amino acid substitutions are shown in Table 7. Amino acid substitutions can be introduced into an antibody of interest and the products screened for a desired activity, e.g., retained / improved antigen binding, reduced immunogenicity, or improved ADCC or CDC. In certain embodiments, the sequences disclosed herein, e.g., CDR sequences, V H Sequence or V L The sequence may have up to about 1, up to about 2, up to about 3, up to about 4, up to about 5, up to about 6, up to about 7, up to about 8, up to about 9, or up to about 10 amino acid residues modified and / or substituted. [Table 7]

[0178] Amino acids can be classified according to general side chain properties: Hydrophobic: Norleucine, Met, Ala, Val, Leu, Ile, ●Neutral hydrophilicity: Cys, Ser, Thr, Asn, Gln, ●Acidic: Asp, Glu, ●Basic: His, Lys, Arg, Residues that affect chain orientation: Gly, Pro, ●Aromatics: Trp, Tyr, Phe.

[0179] Non-conservative substitutions entail exchanging a member of one of these classes for a member of another class.

[0180] 4.3.5. Anti-uPAR Antibodies that Cross-Compete with the Anti-uPAR Antibodies of the Invention for Binding to uPAR The subject matter disclosed herein provides an antibody or antigen-binding fragment thereof that cross-competes with any of the disclosed anti-uPAR antibodies for binding to uPAR (e.g., human uPAR). For example, but not by way of limitation, the cross-competing antibody can bind to the same epitope region, e.g., the same epitope, adjacent epitope, or overlap, as any of the subject matter anti-uPAR antibodies or antigen-binding fragments thereof disclosed herein. In certain embodiments, the reference antibody or reference antigen-binding fragment thereof for cross-competition studies can be any one of the anti-uPAR antibodies or antigen-binding fragments thereof disclosed herein, e.g., 8B1, 11E10, 17C9, 19D7, 6C8, and 14C5 antibodies.

[0181] Such cross-competing antibodies can be identified based on their ability to cross-compete with any one of the anti-uPAR antibodies or antigen-binding fragments thereof disclosed herein in a standard uPAR binding assay. For example, Biacore analysis, ELISA assay or flow cytometry can be used to demonstrate cross-competition with the antibodies of the subject matter disclosed herein. The ability of a test antibody to inhibit, for example, any one of the anti-uPAR antibodies disclosed herein (e.g., 8B1, 11E10, 17C9, 19D7, 6C8, and 14C5 antibodies) binding to uPAR (e.g., human uPAR) demonstrates that the test antibody can compete with any one of the anti-uPAR antibodies or antigen-binding fragments thereof disclosed herein for binding to uPAR (e.g., human uPAR), and thus bind to the same epitope region on uPAR (e.g., human uPAR) as any one of the anti-uPAR antibodies or antigen-binding fragments thereof disclosed herein. In certain embodiments, the cross-competing antibody or antigen-binding fragment thereof binds to the same epitope on uPAR (e.g., human uPAR) as any one of the anti-uPAR antibodies or antigen-binding fragments thereof disclosed herein.

[0182] 4.3.6. Characterization of Antibody Binding to Antigen The subject antibodies or antigen-binding fragments thereof disclosed herein can be tested for binding to uPAR, for example, by standard ELISA. To determine whether selected anti-uPAR antibodies bind to unique epitopes, each antibody can be biotinylated using commercially available reagents (Pierce, Rockford, IL). Competition studies using unlabeled and biotinylated monoclonal antibodies can be performed using uPAR-coated ELISA plates as described above. Biotinylated mAb binding can be detected using a strep-avidin-alkaline phosphatase probe.

[0183] To determine the isotype of purified antibodies, isotype ELISAs can be performed using reagents specific for antibodies of a particular isotype. Anti-uPAR human IgG can be further tested for reactivity with the uPAR antigen by Western blotting.

[0184] In certain embodiments, K D is measured by radiolabeled antigen binding assay (RIA). In certain embodiments, an RIA is performed on a Fab version of the antibody of interest and its antigen. For example, the solution binding affinity of a Fab for an antigen is determined by measuring the binding affinity of the Fab to the antigen at a minimal concentration of ( 125 I) It is measured by equilibrating the Fab with labeled antigen and then capturing the bound antigen with an anti-Fab antibody-coated plate (see, e.g., Chen et al., J Mol Biol (1999); 293:865-881).

[0185] In certain embodiments, K D is measured using a BIACORE® surface plasmon resonance assay, such as an assay using a BIACORE®-2000 or BIACORE®-3000 (BIAcore, Inc., Piscataway, NJ).

[0186] 4.3.7. Immunoconjugates The presently disclosed subject matter provides an anti-uPAR antibody or antigen-binding fragment thereof conjugated to a therapeutic moiety, such as a cytotoxin, a drug (e.g., an immunosuppressant), or a radiotoxin. Such conjugates are referred to herein as "immunoconjugates." Immunoconjugates that include one or more cytotoxins are referred to as "immunotoxins." Cytotoxins or cytotoxic agents include any agent that is detrimental to (e.g., kills) cells. Non-limiting examples of cytotoxins include taxol (such as ricin, diphtheria, gelonin), cytochalasin B, gramicidin D, ethidium bromide, emetine, mitomycin, etoposide, tenoposide, vincristine, vinblastine, colchicine, doxorubicin, daunorubicin, dihydroxyanthracin dione, mitoxantrone, mithramycin, actinomycin D, 1-dehydrotestosterone, glucocorticoids, procaine, tetracaine, lidocaine, propranolol, and puromycin, as well as analogs or homologs thereof. In addition, examples of therapeutic agents include calecheamicin, aureastatin, metabolic antagonists (e.g., methotrexate, 6-mercaptopurine, 6-thioguanine, cytarabine, 5-fluorouracil decarbazine), alkylating agents (e.g., mechlorethamine, thioepaclorambucil, melphalan, carmustine (BSNU) and lomustine (CCNU), cyclophosphamide, busulfan, dibromomannitol, streptozotocin, These include, but are not limited to, cis-dichlorodiamineplatinum(II) (DDP) cisplatin), anthracyclines (e.g., daunorubicin (formerly daunomycin) and doxorubicin), antibiotics (e.g., dactinomycin (formerly actinomycin), bleomycin, mithramycin, and anthramycin (AMC)), hypomethylating agents (azacytidine and decitabine), and antimitotic agents (e.g., vincristine and vinblastine).

[0187] Other examples of therapeutic cytotoxins that can be conjugated to the anti-uPAR antibodies disclosed herein include duocarmycin, calicheamicin, maytansine, and auristatin, and derivatives thereof. Cytotoxins can be conjugated to the anti-uPAR antibodies disclosed herein or antigen-binding fragments thereof using linker technology available in the art. Examples of linker types used to conjugate cytotoxins to antibodies include, but are not limited to, hydrazones, thioethers, esters, disulfides, and peptide-containing linkers. Linkers can be selected that are susceptible to cleavage, for example, by low pH in lysosomal compartments, or by proteases that are preferentially expressed in tumor tissues, such as cathepsins (e.g., cathepsins B, C, D). For further discussion of types of cytotoxins, linkers, and methods for conjugating therapeutic agents to antibodies, see also Saito, G. et al. (2003) Adv. Drug Deliv. Rev. 55:199-215; Trail, PA et al. (2003) Cancer Immunol. Immunother. 52:328-337; Payne, G. (2003) Cancer Cell 3:207-212; Allen, TM (2002) Nat. Rev. Cancer 2:750-763; Pastan, I. and Kreitman, RJ (2002) Curr. Opin. Investig. Drugs 3:1089-1091; Senter, PD and Springer, CJ (2001) Adv. Drug Deliv. Rev. 53:247-264.

[0188] In addition, the anti-uPAR antibody or antigen-binding fragment thereof of the subject matter disclosed herein can be conjugated to an agent that induces senescence. In certain embodiments, the agent that induces senescence is a senogenic agent. Non-limiting examples of senescence-inducing agents include Cdk4 / 6 inhibitors, Cdk2 inhibitors, MEK inhibitors, CDC7 inhibitors, and chemotherapeutic agents. Non-limiting examples of MEK inhibitors include trametinib, cobimetinib, binimetinib, selumetinib, PD-325901, TAK-733, CI-1040 (PD184352), PD0325901, MEK162, AZD8330, GDC-0623, rifametinib, pimasertibi, R04987655, R05126766, WX-554, HL-085, CInQ-03, G-573, PD184161, PD318088, PD98059, R05068760, U0126, and SL327. Non-limiting examples of CDK4 / 6 inhibitors include palbociclib, ribociclib, and abemaciclib. Non-limiting examples of chemotherapeutic agents include cisplatin, doxorubicin, cyclophosphamide, and etoposide.

[0189] The anti-uPAR antibodies or antigen-binding fragments thereof of the presently disclosed subject matter can also be conjugated to radioisotopes to generate cytotoxic radiopharmaceuticals, also called radioimmunoconjugates. Non-limiting examples of radioisotopes that can be conjugated to antibodies for diagnostic or therapeutic use include: 90 Y, 131 I, 225 Ac, 213 Bi, 223 Ra, and 227 Th. Methods for preparing radioimmunoconjugates are established in the art. Examples of radioimmunoconjugates are commercially available, including Zevalin™ (IDEC Pharmaceuticals) and Bexxar™ (Corixa Pharmaceuticals), and similar methods can be used to prepare radioimmunoconjugates using the antibodies of the present invention.

[0190] The anti-uPAR antibodies or antigen-binding fragments thereof of the presently disclosed subject matter can also be conjugated to an imagining agent or probe, for example, for use in imagining techniques, such as ImmunoPET. In certain embodiments, the anti-uPAR antibodies or antigen-binding fragments thereof disclosed herein can be conjugated to an immunoPET probe, such as 9 Zr-Df and 89 It is conjugated to Zr.

[0191] The antibody conjugates of the presently disclosed subject matter can be used to modify a given biological response, and the drug moiety should not be construed as being limited to classical chemotherapeutic agents. For example, the drug moiety can be a protein or polypeptide having a desired biological activity. Such proteins can include, for example, enzymatically active toxins or active fragments thereof (e.g., abrin, ricin A, pseudomonas exotoxin, or diphtheria toxin), proteins (e.g., tumor necrosis factor (TNF) or interferon-γ), or biological response modifiers (e.g., lymphokines, interleukin-1 (IL-1), interleukin-2 (IL-2), interleukin-6 (IL-6), granulocyte macrophage colony stimulating factor (GM-CSF), granulocyte colony stimulating factor (G-CSF), or other growth factors).

[0192] Techniques for conjugating such therapeutic moieties to antibodies are well known and are described, for example, in Arnon et al., "Monoclonal Antibodies For Immunotargeting Of Drugs In Cancer Therapy", Monoclonal Antibodies And Cancer Therapy, Reisfeld et al. (eds.), pp. 243 56 (Alan R. Liss, Inc. 1985); Hellstrom et al., "Antibodies For Drug Delivery", Controlled Drug Delivery (2nd Ed.), Robinson et al. (eds.), pp. 623 53 (Marcel Dekker, Inc. 1987); Thorpe, "Antibody Carriers Of Cytotoxic Agents In Cancer Therapy: A Review", Monoclonal Antibodies '84: Biological And Clinical Applications, Pinchera et al. (eds.), pp. 475 506 (1985); "Analysis, Results, And Future Prospective Of Therapeutic Agents In Cancer Therapy: A Review", Monoclonal Antibodies '84: Biological And Clinical Applications, Pinchera et al. (eds.), pp. 475 506 (1985); See, Therapeutic Use Of Radiolabeled Antibodies In Cancer Therapy”, Monoclonal Antibodies For Cancer Detection And Therapy, Baldwin et al. (eds.), pp. 303 16 (Academic Press 1985), and Thorpe et al., “The Preparation And Cytotoxic Properties Of Antibody-Toxin Conjugates”, Immunol. Rev., 62:119-58 (1982).

[0193] 4.3.8. Multispecific antibodies The subject matter disclosed herein provides a multispecific molecule comprising the anti-uPAR antibody or a fragment thereof disclosed herein. The anti-uPAR antibody or an antigen-binding fragment thereof disclosed herein can be derivatized or linked to one or more functional molecules, such as one or more peptides or proteins (e.g., one or more antibodies or ligands for a receptor), to generate a multispecific molecule that binds to two or more different binding sites or target molecules. The anti-uPAR antibody or an antigen-binding fragment thereof disclosed herein can in fact be derivatized or linked to two or more other functional molecules to generate a multispecific molecule that binds to three or more different binding sites and / or target molecules. To generate a multispecific molecule, the anti-uPAR antibody or an antigen-binding fragment thereof disclosed herein can be functionally linked (e.g., by chemical coupling, genetic fusion, non-covalent association, or other methods) to one or more other binding molecules, such as another antibody, antibody fragment, peptide, or binding mimic, to result in a multispecific molecule.

[0194] In certain embodiments, the multispecific molecule is a bispecific molecule. In certain embodiments, the bispecific molecule comprises at least a first binding specificity for uPAR and a second binding specificity for a second target epitope region. The second target epitope region can be a uPAR epitope or a non-uPAR epitope, e.g., a different antigen. In certain embodiments, the multispecific molecule comprises a first binding specificity for uPAR, a second binding specificity for a second target, and a third binding specificity for a third target. In certain embodiments, the second target is an antigen expressed on the surface of an immune cell (e.g., a T cell, or a human immune effector cell). In certain embodiments, the multispecific molecule can specifically bind to an effector antigen on a human immune effector cell, thereby recruiting the activity of the immune effector cell and thereby enhancing the effector function. In certain embodiments, the third target is an antigen expressed on a senescent cell.

[0195] The multispecific molecules of the presently disclosed subject matter can be prepared by conjugating the component binding specificities using methods known in the art. For example, each binding specificity for a multispecific molecule can be generated separately and then conjugated to each other. When the binding specificities are proteins or peptides, a variety of coupling or cross-linking agents can be used for covalent conjugation. Non-limiting examples of cross-linking agents include protein A, carbodiimide, N-succinimidyl-S-acetyl-thioacetate (SATA), 5,5'-dithiobis(2-nitrobenzoic acid) (DTNB), o-phenylenedimaleimide (oPDM), N-succinimidyl-3-(2-pyridyldithio)propionate (SPDP), and sulfosuccinimidyl 4-(N-maleimidomethyl)cyclohexane-1-carboxylate (sulfo-SMCC) (see, e.g., Karpovsky et al. (1984) J. Exp. Med. 160:1686; Liu, MA et al. (1985) Proc. Natl. Acad. Sci. USA 82:8648). Other methods include those of Paulus (1985) Behring Ins. Mitt. No. 78, 118-132; Brennan et al. (1985) Science 229:81-83), and Glennie et al. (1987) J. Immunol. 139:2367-2375. Conjugating agents can be SATA and sulfo-SMCC, both available from Pierce Chemical Co. (Rockford, IL).

[0196] When the binding specificities are antibodies, they can be conjugated via sulfhydryl bonds in the C-terminal hinge regions of the two heavy chains. In certain embodiments, the hinge region is modified to contain an odd number of sulfhydryl residues, preferably one, prior to conjugation.

[0197] Alternatively, both binding specificities can be encoded in the same vector and expressed and assembled in the same host cell, this method is particularly useful when the multispecific molecule is a mAb x mAb, mAb x Fab, F(ab')2, or ligand x Fab fusion protein.

[0198] Binding of multispecific molecules to their specific targets can be confirmed, for example, by enzyme-linked immunosorbent assay (ELISA), radioimmunoassay (RIA), FACS analysis, bioassay (e.g., growth inhibition), or Western blot assay. Each of these assays generally detects the presence of a particular protein-antibody complex of interest by using a labeled reagent (e.g., an antibody) specific for the complex of interest. Alternatively, the complex can be detected using any of a variety of other immunoassays. For example, the antibody can be radioactively labeled and used in a radioimmunoassay (RIA) (see, for example, Weintraub, B., Principles of Radioimmunoassays, Seventh Training Course on Radioligand Assay Techniques, The Endocrine Society, March, 1986, incorporated herein by reference). Radioisotopes can be detected by such means as the use of a gamma counter or a scintillation counter, or by autoradioassay testing.

[0199] 4.4. Nucleic Acids Encoding Antibodies or Antigen-Binding Fragments The presently disclosed subject matter provides nucleic acids encoding the anti-uPAR antibodies, or antigen-binding fragments thereof, disclosed herein.

[0200] Further provided is a vector comprising the nucleic acid disclosed herein. In some embodiments, the vector is an expression vector. The subject matter disclosed herein further provides a host cell comprising the vector disclosed herein. In certain embodiments, the host cell is a T cell.

[0201] 4.5. Pharmaceutical Compositions and Methods of Treatment The subject matter disclosed herein provides a composition comprising the anti-uPAR antibody or antigen-binding fragment thereof disclosed herein, the immunoconjugate disclosed herein, or the multispecific molecule disclosed herein. In one embodiment, the composition is a pharmaceutical composition further comprising a pharma- ceutically acceptable carrier.

[0202] The anti-uPAR antibody or its antigen-binding fragment of the subject matter disclosed herein can be administered in the form of a composition further comprising a pharmaceutically acceptable carrier.Suitable pharmaceutically acceptable carriers include, for example, one or more of water, saline, phosphate buffered saline, dextrose, glycerol, ethanol, etc., and combinations thereof.

[0203] Pharmaceutically acceptable carriers may further contain minor amounts of auxiliary substances such as wetting or emulsifying agents, preservatives, or buffers, which enhance the shelf life or effectiveness of the binding protein.The compositions for injection can be formulated so as to provide quick, sustained, or delayed release of the active ingredient after administration to a mammal, as is well known to those skilled in the art.

[0204] The subject matter disclosed herein provides various methods of using the anti-uPAR antibodies or antigen-binding fragments thereof, immunoconjugates, multispecific molecules, and compositions disclosed herein in therapy. The subject matter disclosed herein provides methods for treating or alleviating a disease or disorder in a subject. In some embodiments, the disease or disorder is associated with uPAR. In certain embodiments, the disease or disorder is associated with overexpression of uPAR. In certain embodiments, the disease or disorder is selected from the group consisting of tumors, aging-related pathologies, and age-related tissue loss. Non-limiting examples of aging-related pathologies include pulmonary fibrosis, atherosclerosis, Alzheimer's disease, diabetes, osteoarthritis, liver fibrosis, chronic kidney disease, cardiac fibrosis, and Parkinson's disease.

[0205] In certain embodiments, the method comprises administering to a subject in need thereof an anti-uPAR antibody or antigen-binding fragment thereof, immunoconjugate, multispecific molecule, or composition disclosed herein.

[0206] For purposes of treatment, the amount of anti-uPAR antibody or antigen-binding fragment thereof, immunoconjugate, or multispecific molecule provided herein administered is an amount effective to produce the desired effect, e.g., treatment or mitigation of a disease or disorder (e.g., tumors and aging-related pathologies). An effective amount can be provided in a single administration or a series of administrations of the anti-uPAR antibody or antigen-binding fragment thereof, immunoconjugate, or multispecific molecule disclosed herein.

[0207] The anti-uPAR antibodies or antigen-binding fragments thereof, immunoconjugates, and multispecific molecules of the subject matter disclosed herein may be administered by any method known in the art, including, but not limited to, pleural, intravenous, subcutaneous, intranodal, intratumoral, intrathecal, intravitreal, intrapleural, intraperitoneal, and direct administration into the thymus.

[0208] In certain embodiments, the disease or disorder is a tumor. In certain embodiments, the anti-uPAR antibody or antigen-binding fragment thereof, immunoconjugate, or multispecific molecule disclosed herein can reduce tumor burden in a subject, reduce the number of tumor cells, reduce tumor size, and / or eradicate the tumor, and / or increase or prolong the survival of the subject.

[0209] Non-limiting examples of tumors include breast cancer (including triple-negative breast cancer), endometrial cancer, ovarian cancer, colon cancer, rectal cancer, lung cancer (e.g., non-small cell lung cancer), stomach cancer, prostate cancer, gastric cancer, renal cancer, pancreatic cancer, blood cancer, cervical cancer, head and neck cancer, liver cancer (e.g., cholangiocarcinoma, hepatocellular carcinoma, and fibrolamaellar hepatocellular carcinoma), urothelial carcinoma, melanoma, and brain cancer (including glioblastoma multiforme). In certain embodiments, the blood cancer is selected from the group consisting of acute lymphoblastic leukemia (ALL), chronic lymphocytic leukemia (CLL), acute myeloid leukemia (AML), myelofibrosis, polycythemia vera, myelodysplastic syndrome, and erythroleukemia. In certain embodiments, the tumor is cancer. In certain embodiments, the cancer is a recurrent or refractory cancer. In certain embodiments, the cancer is resistant to a cancer therapy, for example, chemotherapy.

[0210] Furthermore, the subject matter disclosed herein provides a method for increasing the production of immune-activating cytokines in response to tumor cells in a subject. In certain embodiments, the method comprises administering to the subject an anti-uPAR antibody or antigen-binding fragment thereof, immunoconjugate, or multispecific molecule disclosed herein. Non-limiting examples of immune-activating cytokines include granulocyte-macrophage colony-stimulating factor (GM-CSF), IFN-α, IFN-β, IFN-γ, TNF-α, IL-1, IL-2, IL-3, IL-6, IL-11, IL-7, IL-8, IL-12, IL-15, IL-21, interferon regulatory factor 7 (IRF7), CCL1, CCL2, CCL3, CCL5, CCL7, CCL8, CCL13, CCL16, CXCL1, CXCL3, CXCL5, CXCL9, CXCL10, and combinations thereof.

[0211] In certain embodiments, the disease or disorder is a senescence-associated pathology. In certain embodiments, the subject exhibits increased accumulation of senescent cells compared to cells observed in healthy control subjects. In certain embodiments, the senescence-associated pathology is selected from the group consisting of pulmonary fibrosis, atherosclerosis, Alzheimer's disease, diabetes, liver fibrosis, chronic kidney disease, osteoarthritis, cardiac fibrosis, and Parkinson's disease. In certain embodiments, the senescent cells exhibit senescence-associated secretory phenomena (SASP). The senescence-associated secretory phenomena may be induced by replication, oncogenes (e.g., HRASG12D, NRAsG12D, NRAsG12D, etc.), radiation, chemotherapy, or drugs (e.g., Cdk4 / 6 inhibitors, MEK inhibitors, chemotherapeutic drugs, etc.). Non-limiting examples of MEK inhibitors include trametinib, cobimetinib, binimetinib, selumetinib, PD-325901, TAK-733, CI-1040 (PD184352), PD0325901, MEK162, AZD8330, GDC-0623, rifametinib, pimasertibi, R04987655, R05126766, WX-554, HL-085, CInQ-03, G-573, PD184161, PD318088, PD98059, R05068760, U0126, and SL327. Non-limiting examples of CDK4 / 6 inhibitors include palbociclib, ribociclib, and abemaciclib. Non-limiting examples of chemotherapeutic agents include cisplatin, doxorubicin, cyclophosphamide, and etoposide.

[0212] In certain embodiments, the methods disclosed herein for treating or alleviating a disease or disorder further comprise administering a tumor-specific monoclonal antibody to a subject, and the subject is undergoing / has undergone a senescence-inducing therapy (e.g., chemotherapy). In certain embodiments, the tumor-specific monoclonal antibody is administered after administration of the anti-uPAR antibody or antigen-binding fragment thereof, immunoconjugate, or multispecific molecule. Non-limiting examples of senescence-inducing therapy include doxorubicin, ionizing radiation therapy, combination therapy with MEK inhibitors and CDK4 / 6 inhibitors, combination therapy with CDC7 inhibitors and mTOR inhibitors, etc. Examples of CDK4 / 6 inhibitors include palbociclib, ribociclib, and abemaciclib. Non-limiting examples of MEK inhibitors include trametinib, cobimetinib, binimetinib, selumetinib, PD-325901, TAK-733, CI-1040 (PD184352), PD0325901, MEK162, AZD8330, GDC-0623, rifametinib, pimasertibi, R04987655, R05126766, WX-554, HL-085, CInQ-03, G-573, PD184161, PD318088, PD98059, R05068760, U0126, and SL327. Non-limiting examples of mTOR inhibitors include rapamycin, sertraline, sirolimus, everolimus, temsirolimus, ridaforolimus, and deforolimus. Examples of CDC7 inhibitors include TAK-931, PHA-767491, XL413, 1H-pyrrolo[2,3-b]pyridine, 2,3-dihydrothieno[3,2-d]pyrimidin-4(1H)-one, furanone derivatives, trisubstituted thiazoles, pyrrolopyridinones, and the like.

[0213] In certain embodiments, the tumor-specific monoclonal antibody is administered following administration of the anti-uPAR antibody or antigen-binding fragment thereof, immunoconjugate, or multispecific molecule.

[0214] In certain embodiments, the subject is a human.

[0215] In certain embodiments, the methods disclosed herein for treating or alleviating a disease or disorder further comprise administering to the subject a cancer therapy, hi certain embodiments, the cancer therapy is selected from the group consisting of chemotherapy, radiation therapy, immunotherapy, monoclonal antibodies, anti-cancer nucleic acids or proteins, anti-cancer viruses or microbes, and any combination thereof.

[0216] In certain embodiments, the methods disclosed herein for treating or alleviating a disease or disorder further comprise administering to the subject a cytokine, hi certain embodiments, the cytokine is administered before, during, or after administration of the anti-uPAR antibody or antigen-binding fragment thereof, immunoconjugate, or multispecific molecule. In certain embodiments, the cytokine is interferon a, interferon (3, interferon y, complement C5a, IL-2, TNF-α, CD4OL, IL12, IL-23, IL15, IL17, CCL1, CCL11, CCL12, CCL13, CCL14-1, CCL14-2, CCL14-3, CCL15-1, CCL15-2, CCL16, CCL17, CCL18, CCL19, CCL19, CCL2, CCL20, CCL21, CCL22, CCL23-1, CCL23-2, CCL24, CCL25-1, CCL25-2, CCL26, CCL27, CCL28, CCL29, CCL30, CCL31, CCL32, CCL33-1, CCL33-2, CCL34, CCL35-1, CCL35-2, CCL36, CCL37, CCL38, CCL39, CCL40, CCL41, CCL42, CCL43-2, CCL44, CCL45-1, CCL45-2, CCL46, CCL47, CCL48, CCL49, CCL50, CCL51, CCL52, CCL53, CCL54, CCL55, CCL56, CCL57, CCL58, CCL59, CCL60, CCL61, CCL62, CCL63, CCL64, CCL65, CCL66, CCL67, CCL68, CCL69, CCL70, CCL71, CCL72, CCL73, CCL74, CCL75, CCL75, CCL76, CCL77, CCL78, CCL79, CCL80, CCL81, CCL CL3, CCL3L1, CCL4, CCL4L1, CCL5, CCL6, CCL7, CCL8, CCL9, CCR10, CCR2, CCR5, CCR6, CCR7, CCR8, CCRL1, CCRL2, CX3CL1, CX3CR, CXCL1, CXCL10, CXCL11, CXCL12, CXCL13, CXCL14, CXCL15, CXCL16, CXCL2, CXCL3, CXCL4, CXCL5, CXCL6, CXCL7, CXCL8, CXCL9, CXCL9, CXCR1, CXCR2, CXCR4, CXCR5, CXCR6, CXCR7, and XCL2.

[0217] In certain embodiments, chemotherapy comprises administering to the subject a chemotherapeutic agent, non-limiting examples of which include nitrogen mustards, ethylenimine derivatives, alkylsulfonates, nitrosoureas, gemcitabine, triazenes, folic acid analogs, anthracyclines, taxanes, COX-2 inhibitors, pyrimidine analogs, purine analogs, antibiotics, enzyme inhibitors, epipodophyllotoxins, platinum coordination complexes, vinca alkaloids, substituted ureas, methylhydrazine derivatives, adrenocortical suppressants, hormone antagonists, endostatin, taxol, camptothecin, SN-38, doxorubicin, doxorubicin analogs, antimetabolites, alkylating agents, antimitotic agents, antiangiogenic agents, tyrosine kinase inhibitors, mTOR inhibitors, heat shock protein (HSP90) inhibitors, proteosome inhibitors, HDAC inhibitors, pro-apoptotic agents, methotrexate, and CPT-11.

[0218] In certain embodiments, the disease or disorder is pulmonary fibrosis, and the method further comprises administering to the subject at least one therapy selected from the group consisting of pirfenidone, nintedanib, oxygen therapy, corticosteroids (e.g., prednisone), mycophenolate mofetil / mycophenolic acid, and azathioprine, either sequentially, separately, or simultaneously.

[0219] In certain embodiments, the disease or disorder is atherosclerosis, and the method further comprises administering to the subject at least one therapy selected from the group consisting of statins (e.g., atorvastatin, fluvastatin, lovastatin, pitavastatin, pravastatin, rosuvastatin calcium, simvastatin), fibrates (e.g., gemfibrozil, fenofibrate), niacin, ezetimibe, bile acid sequestrants (e.g., cholestyramine, colestipol, colesevelam), proprotein convertase subtilisin kexin type 9 (PCSK9) inhibitors, antiplatelet agents (e.g., aspirin, clopidogrel, tacagrelor, warfarin, prasugrel), beta blockers, angiotensin-converting enzyme (ACE) inhibitors, calcium channel blockers, and diuretics, either sequentially, separately, or simultaneously.

[0220] In certain embodiments, the disease or disorder is Alzheimer's disease, and the method further comprises administering to the subject at least one therapy selected from the group consisting of donepezil, galantamine, memantine, rivastigmine, memantine extended release and donepezil (Namzaric), aducanumab, solanezumab, insulin, verubecestat, AADvac1, CSP-1103, and intepirdine, either sequentially, separately, or simultaneously.

[0221] In certain embodiments, the disease or disorder is diabetes, and the method includes administering to the subject insulin, metformin, an amylin analog, glucagon, a sulfonylurea (e.g., glimepiride, glipizide, glyburide, chlorpropamide, tolazamide, tolbutamide), a meglitinide (e.g., nateglinide, repaglinide), a thiazolidinedione (e.g., pioglitazone, rosiglitazone), an alpha-glucosidase inhibitor (e.g., acarbose, miglitol), a dipeptidyl peptidase (DPP-4 The method further comprises administering, sequentially, separately or simultaneously, at least one therapy selected from the group consisting of: sodium-glucose cotransporter 2 (SGLT2) inhibitors (e.g., alogliptin, linagliptin, sitagliptin, saxagliptin), sodium-glucose cotransporter 2 (SGLT2) inhibitors (e.g., canagliflozin, dapagliflozin, empagliflozin, ertugliflozin), and incretin mimetics (e.g., exenatide, liraglutide, dulaglutide, lixisenatide, semaglutide).

[0222] In certain embodiments, the disease or disorder is osteoarthritis, and the method further comprises administering to the subject, sequentially, separately, or simultaneously, at least one therapy selected from the group consisting of analgesics (e.g., acetaminophen, tramadol, oxycodone, hydrocodone), nonsteroidal anti-inflammatory drugs (e.g., aspirin, ibuprofen, naproxen, celecoxib), cyclooxygenase-2 inhibitors, corticosteroids, and hyaluronic acid.

[0223] In certain embodiments, the disease or disorder is liver fibrosis, and the method further comprises administering to the subject, sequentially, separately, or simultaneously, at least one therapy selected from the group consisting of ACE inhibitors (e.g., benazepril, lisinopril, ramipril), α-tocopherol, interferon-α, PPAR antagonists, colchicine, corticosteroids, endothelin inhibitors, interleukin-10, pentoxifylline, phosphatidylcholine, S-adenosyl-methionine, and TGF-131 inhibitors.

[0224] In certain embodiments, the disease or disorder is chronic kidney disease and the method further comprises administering to the subject, sequentially, separately, or simultaneously, at least one therapy selected from the group consisting of an ACE inhibitor (e.g., benazepril, lisinopril, ramipril), a statin (e.g., atorvastatin, fluvastatin, lovastatin, pitavastatin, pravastatin, rosuvastatin calcium, simvastatin), furosemide, erythropoietin, a phosphate binder (e.g., calcium acetate, calcium carbonate), cholecalciferol, ergocalciferol, and cyclophosphamide.

[0225] 4.6. Diagnostic and prognostic methods The anti-uPAR antibodies, antigen-binding fragments thereof, multispecific molecules, and their nucleic acid codes disclosed herein can be used for diagnostic and prognostic applications, as well as for use as research tools for the detection of uPAR in biological samples, cells, tissues, and / or blood samples. The subject matter disclosed herein provides a method for detecting uPAR in a cell, tissue, or blood sample. In certain embodiments, the method includes contacting a cell or tissue with an antibody, antigen-binding fragment thereof, or multispecific molecule disclosed herein, where the antibody, antigen-binding fragment thereof, or multispecific molecule comprises a detectable label, and determining the amount of the labeled antibody, antigen-binding fragment thereof, or multispecific molecule bound to the cell, tissue, or blood sample by measuring the amount of detectable label associated with the cell or tissue, where the amount of bound antibody, antigen-binding fragment thereof, or multispecific molecule indicates the amount of uPAR in the cell, tissue, or blood sample. The cell or tissue can be any cell or tissue, including any normal, healthy, or cancerous cell and tissue. In certain embodiments, the blood sample is a peripheral blood sample.

[0226] uPAR can be used as a marker for detecting senescent cell burden in a subject. Thus, the antibodies, antigen-binding fragments thereof, or multispecific molecules disclosed herein can be used to detect senescent cells in a biological sample obtained from a subject. The subject matter disclosed herein provides a method for detecting senescent cells in a biological sample obtained from a subject. In certain embodiments, the method comprises: a) contacting a biological sample with an antibody, antigen-binding fragment thereof, or multispecific molecule disclosed herein, where the antibody, antigen-binding fragment thereof, or multispecific molecule comprises a detectable label; b) determining the amount of labeled antibody, antigen-binding fragment thereof, or multispecific molecule in the biological sample by measuring the amount of detectable label in the biological sample, where the amount of bound antibody, antigen-binding fragment thereof, or multispecific molecule indicates the amount of uPAR in the biological sample; and c) determining a level of uPAR that is at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55% relative to that observed in i) a reference sample. , at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, or at least 100% increased, and / or ii) at least 0.5-fold, at least 1.0-fold, at least 1.5-fold, at least 2.0-fold, at least 2.5-fold, at least 3.0-fold, at least 3.5-fold, at least 4.0-fold, at least 4.5-fold, at least 5.0-fold, at least 5.5-fold, at least 6.0-fold, at least 6.5-fold, at least 7.0-fold, at least 7.5-fold, at least 8.0-fold, at least 8.5-fold, at least 9.0-fold, at least 9.5-fold, or at least 10.0-fold increased in the biological sample compared to that observed in the reference sample. The detectable label can be an immunoPET probe. The reference sample can be obtained from a healthy control subject or can contain a predetermined level of uPAR polypeptide and / or suPAR polypeptide.Non-limiting examples of biological samples include mucus, saliva, bronchoalveolar lavage fluid (BAL), bronchial washings (BW), whole blood, cerebrospinal fluid (CSF), urine, plasma, serum, lymph, semen, synovial fluid, tears, amniotic fluid, bile, aqueous humor, and body fluids. In certain embodiments, measuring the amount of detectable label in a biological sample includes Western blotting, flow cytometry, enzyme-linked immunosorbent assay (ELISA), immunoprecipitation, immunoelectrophoresis, immunostaining, imaging techniques (e.g., immunoPET), isoelectric focusing, high performance liquid chromatography (HPLC), or mass spectrometry.

[0227] 4.7.Kit The subject matter disclosed herein provides a kit for treating or alleviating disease or disorder and / or detecting uPAR.In certain embodiments, the kit comprises the anti-uPAR antibody or its antigen-binding fragment, immunoconjugate, multispecific molecule, or composition disclosed herein.In certain embodiments, the kit comprises a sterile container that contains a therapeutic or prophylactic vaccine, and such container can be a box, an ampoule, a bottle, a vial, a tube, a bag, a pouch, a blister pack, or other suitable container form known in the art.Such container can be made of plastic, glass, laminated paper, metal foil, or other material suitable for holding pharmaceutical products.

[0228] In certain embodiments, the kit further comprises instructions for administering the anti-uPAR antibody or antigen-binding fragment thereof, immunoconjugate, multispecific molecule, or composition disclosed herein to a subject in need of treatment. The instructions can generally include information regarding the use of the anti-uPAR antibody or antigen-binding fragment thereof, immunoconjugate, multispecific molecule, and composition disclosed herein for the treatment or alleviation of a disease or disorder. In certain embodiments, the instructions include at least one of the following: description of the therapeutic agent, dosing schedule and administration for the treatment and / or prevention of a tumor or neoplasm or symptoms thereof, precautions, warnings, indications, counterindications, overdose information, adverse reactions, animal pharmacology, clinical studies, and / or references. The instructions can be printed directly on the container (if present), or as a label applied to the container, or as a separate sheet, pamphlet, card, or folder provided in or with the container.

[0229] 4.8. Illustrative Embodiments A. In certain non-limiting embodiments, the subject matter disclosed herein provides an anti-uPAR antibody or antigen-binding fragment thereof comprising a heavy chain variable region comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or at least about 100% homologous or identical to the amino acid sequence set forth in SEQ ID NO:25, SEQ ID NO:27, SEQ ID NO:29, SEQ ID NO:31, SEQ ID NO:47, or SEQ ID NO:55.

[0230] A1. An anti-uPAR antibody or its antigen-binding fragment according to A above, wherein the heavy chain variable region comprises an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or at least about 100% homologous or identical to the amino acid sequence set forth in SEQ ID NO:27.

[0231] A2. In certain non-limiting embodiments, the subject matter disclosed herein provides an anti-uPAR antibody or antigen-binding fragment thereof comprising a light chain variable region comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or at least about 100% homologous or identical to the amino acid sequence set forth in SEQ ID NO:26, SEQ ID NO:28, SEQ ID NO:30, SEQ ID NO:32, SEQ ID NO:48, or SEQ ID NO:56.

[0232] A3. The anti-uPAR antibody or its antigen-binding fragment of A2 described above, wherein the light chain variable region comprises an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or at least about 100% homologous or identical to the amino acid sequence set forth in SEQ ID NO:28.

[0233] A4. In certain non-limiting embodiments, the subject matter disclosed herein provides an anti-uPAR antibody or antigen-binding fragment thereof comprising: (a) a heavy chain variable region comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or at least about 100% homologous or identical to the amino acid sequence set forth in SEQ ID NO:25, SEQ ID NO:27, SEQ ID NO:29, SEQ ID NO:31, SEQ ID NO:47, or SEQ ID NO:55; and (b) a light chain variable region comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or at least about 100% homologous or identical to the amino acid sequence set forth in SEQ ID NO:26, SEQ ID NO:28, SEQ ID NO:30, SEQ ID NO:32, SEQ ID NO:48, or SEQ ID NO:56.

[0234] A5. In certain non-limiting embodiments, the subject matter disclosed herein is an anti-uPAR antibody or antigen-binding fragment thereof comprising a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region and the light chain variable region comprise: (a) a heavy chain variable region comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, at least about 100% homologous or identical to the amino acid sequence set forth in SEQ ID NO:25; and (b) a heavy chain variable region comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, at least about 100% homologous or identical to the amino acid sequence set forth in SEQ ID NO:26. (b) a light chain variable region comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or at least about 100% homologous or identical to the amino acid sequence set forth in SEQ ID NO:27; and (c) a heavy chain variable region comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or at least about 100% homologous or identical to the amino acid sequence set forth in SEQ ID NO:27; and (c) a light chain variable region comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or at least about 100% homologous or identical to the amino acid sequence set forth in SEQ ID NO:29; (d) a heavy chain variable region comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or at least about 100% homologous or identical to the amino acid sequence set forth in SEQ ID NO:31;(e) a heavy chain variable region comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, at least about 100% homologous or identical to the amino acid sequence set forth in SEQ ID NO: 32; (f) a light chain variable region comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, at least about 100% homologous or identical to the amino acid sequence set forth in SEQ ID NO: 47; and and (f) a heavy chain variable region comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, at least about 100% homologous or identical to the amino acid sequence set forth in SEQ ID NO:55, and a light chain variable region comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, at least about 100% homologous or identical to the amino acid sequence set forth in SEQ ID NO:56.

[0235] A6. The anti-uPAR antibody or its antigen-binding fragment of A5 described above, wherein the heavy chain variable region comprises an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or at least about 100% homologous or identical to the amino acid sequence set forth in SEQ ID NO:27, and the light chain variable region comprises an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or at least about 100% homologous or identical to the amino acid sequence set forth in SEQ ID NO:28.

[0236] A7. In certain non-limiting embodiments, the subject matter disclosed herein provides an anti-uPAR antibody or antigen-binding fragment thereof comprising a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO:25, SEQ ID NO:27, SEQ ID NO:29, SEQ ID NO:31, SEQ ID NO:47, or SEQ ID NO:55.

[0237] A8. The anti-uPAR antibody or antigen-binding fragment thereof of A7 above, wherein the heavy chain variable region comprises the amino acid sequence set forth in SEQ ID NO:27.

[0238] A9. In certain non-limiting embodiments, the subject matter disclosed herein provides an anti-uPAR antibody or antigen-binding fragment thereof comprising a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:26, SEQ ID NO:28, SEQ ID NO:30, SEQ ID NO:32, SEQ ID NO:48, or SEQ ID NO:56.

[0239] A10. The anti-uPAR antibody or antigen-binding fragment thereof of A9 above, wherein the light chain variable region comprises the amino acid sequence set forth in SEQ ID NO:28.

[0240] A11. In certain non-limiting embodiments, the subject matter disclosed herein provides an anti-uPAR antibody or antigen-binding fragment thereof comprising: (a) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO:25, SEQ ID NO:27, SEQ ID NO:29, SEQ ID NO:31, SEQ ID NO:47, or SEQ ID NO:55; and (b) a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:26, SEQ ID NO:28, SEQ ID NO:30, SEQ ID NO:32, SEQ ID NO:48, or SEQ ID NO:56.

[0241] A12. Any one of the antibodies A to A11 above, or an antigen-binding fragment thereof, comprising: (a) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 25, and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 26; (b) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 27, and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 28; (c) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 29, and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 30; (d) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 31, and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 32; (e) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 47, and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 48; and (f) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 55, and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 56.

[0242] A13. The antibody or antigen-binding fragment thereof according to the above A12, wherein the heavy chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 27, and the light chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 28.

[0243] A14. In certain non-limiting embodiments, the subject matter disclosed herein is an anti-uPAR antibody or antigen-binding fragment thereof comprising a heavy chain variable region comprising a CDR1 domain, a CDR2 domain, and a CDR3 domain, and a light chain variable region comprising a CDR1 domain, a CDR2 domain, and a CDR3 domain, wherein the CDR3 domain of the heavy chain variable region and the light chain variable region comprises: (a) a heavy chain variable region CDR3, CDR3 comprising the amino acid sequence set forth in SEQ ID NO:3 and a conservative modification thereof, and a light chain variable region CDR3, CDR3 comprising the amino acid sequence set forth in SEQ ID NO:6 and a conservative modification thereof; (b) a heavy chain variable region CDR3 comprising the amino acid sequence set forth in SEQ ID NO:9 and a conservative modification thereof, and a light chain variable region CDR3 comprising the amino acid sequence set forth in SEQ ID NO:12 and a conservative modification thereof; (c) an amino acid sequence set forth in SEQ ID NO:15 and a conservative modification thereof; and a conservative modification thereof, and a light chain variable region CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 18 and a conservative modification thereof; (d) a heavy chain variable region CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 21 and a conservative modification thereof, and a light chain variable region CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 24 and a conservative modification thereof; (e) a heavy chain variable region CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 43 and a conservative modification thereof, and a light chain variable region CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46 and a conservative modification thereof; and (f) a heavy chain variable region CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 51 and a conservative modification thereof, and a light chain variable region CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 54 and a conservative modification thereof.

[0244] A15. The CDR2 domains of the heavy chain variable region and the light chain variable region are (a) a heavy chain variable region CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 2 and a conservative modification thereof, and a light chain variable region CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 5 and a conservative modification thereof, (b) a heavy chain variable region CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 8 and a conservative modification thereof, and a light chain variable region CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 11 and a conservative modification thereof, (c) a heavy chain variable region CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 14 and a conservative modification thereof, and a light chain variable region CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 17 and a conservative modification thereof, The antibody or antigen-binding fragment thereof of the aforementioned A14, selected from: (d) a heavy chain variable region CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 20 and a conservative modification thereof, and a light chain variable region CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 23 and a conservative modification thereof; (e) a heavy chain variable region CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 42 and a conservative modification thereof, and a light chain variable region CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 45 and a conservative modification thereof; and (f) a heavy chain variable region CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 50 and a conservative modification thereof, and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 53 and a conservative modification thereof.

[0245] A16. The CDR1 domains of the heavy chain variable region and the light chain variable region are (a) a heavy chain variable region CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 1 and a conservative modification thereof, and a light chain variable region CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 4 and a conservative modification thereof, (b) a heavy chain variable region CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 7 and a conservative modification thereof, and a light chain variable region CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 10 and a conservative modification thereof, (c) a heavy chain variable region CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 13 and a conservative modification thereof, and a light chain variable region CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 16 and a conservative modification thereof, (d) a heavy chain variable region CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 17 and a conservative modification thereof, and a light chain variable region CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 19 and a conservative modification thereof, and a light chain variable region CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 20 and a conservative modification thereof, and a heavy chain variable region CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 21 and a conservative modification thereof, and a light chain variable region CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 22 and a conservative modification thereof, and a light chain variable region CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 23 and a conservative modification thereof, and a light chain variable region CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 24 and a conservative modification thereof, and a The antibody or antigen-binding fragment thereof of A14 or A15 described above, selected from: (a) a heavy chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 19 and a conservative modification thereof, and a light chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 22 and a conservative modification thereof; (b) a heavy chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 41 and a conservative modification thereof, and a light chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 44 and a conservative modification thereof; and (c) a heavy chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 49 and a conservative modification thereof, and a light chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 52 and a conservative modification thereof.

[0246] A17. An antibody or an antigen-binding fragment thereof according to any one of the above A14 to A16, wherein one or more of the CDR sequences have up to about 5 amino acid substitutions.

[0247] A18. An antibody or an antigen-binding fragment thereof according to any one of the above A14 to A16, wherein one or more of the CDR sequences have up to about 3 amino acid substitutions.

[0248] A19. In certain non-limiting embodiments, the subject matter disclosed herein comprises a CDR1 having an amino acid sequence as set forth in SEQ ID NO:1, a CDR2 having an amino acid sequence as set forth in SEQ ID NO:2, and a CDR3 having an amino acid sequence as set forth in SEQ ID NO:3; (b) a CDR1 having an amino acid sequence as set forth in SEQ ID NO:7, a CDR2 having an amino acid sequence as set forth in SEQ ID NO:8, and a CDR3 having an amino acid sequence as set forth in SEQ ID NO:9; (c) a CDR1 having an amino acid sequence as set forth in SEQ ID NO:13, a CDR2 having an amino acid sequence as set forth in SEQ ID NO:14, and a CDR3 having an amino acid sequence as set forth in SEQ ID NO:15; (d) a CDR1 having an amino acid sequence as set forth in SEQ ID NO:16, a CDR2 having an amino acid sequence as set forth in SEQ ID NO:17, and a CDR3 having an amino acid sequence as set forth in SEQ ID NO:19; The present invention provides an anti-uPAR antibody or an antigen-binding fragment thereof, comprising a heavy chain variable region comprising: (a) CDR1 having the amino acid sequence set forth in sequence number 19, a CDR2 having the amino acid sequence set forth in sequence number 20, and a CDR3 having the amino acid sequence set forth in sequence number 21; (b) CDR1 having the amino acid sequence set forth in sequence number 41, a CDR2 having the amino acid sequence set forth in sequence number 42, and a CDR3 having the amino acid sequence set forth in sequence number 43; or (c) CDR1 having the amino acid sequence set forth in sequence number 49, a CDR2 having the amino acid sequence set forth in sequence number 50, and a CDR3 having the amino acid sequence set forth in sequence number 51.

[0249] A20. In certain non-limiting embodiments, the subject matter disclosed herein comprises: (a) a CDR1 comprising the amino acid sequence set forth in SEQ ID NO:4, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO:5, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO:6; (b) a CDR1 comprising the amino acid sequence set forth in SEQ ID NO:10, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO:11, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO:12; (c) a CDR1 comprising the amino acid sequence set forth in SEQ ID NO:16, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO:17, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO:18; (d) a CDR1 comprising the amino acid sequence set forth in SEQ ID NO:19, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO:20; ) an anti-uPAR antibody or an antigen-binding fragment thereof comprising a light chain variable region comprising: (a) CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 22, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 23, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 24; (b) a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 44, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 45, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46; or (c) a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 52, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 53, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 54.

[0250] A21. In certain non-limiting embodiments, the subject matter disclosed herein comprises: (a) a heavy chain variable region comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO:1, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO:2, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO:3; and a light chain variable region comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO:4, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO:5, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO:6; (b) a light chain variable region comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO:7, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO:8; and CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 9; and a light chain variable region comprising CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 10, CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 11, and CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 12; (c) a heavy chain variable region comprising CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 13, CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 14, and CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 15, and CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 16, and (d) a light chain variable region comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 19, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 20, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 21, and a light chain variable region comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 22, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 23, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 24; and (e) a light chain variable region comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 41. a heavy chain variable region comprising CDR1 comprising the amino acid sequence of SEQ ID NO: 43, CDR2 comprising the amino acid sequence of SEQ ID NO: 44, CDR2 comprising the amino acid sequence of SEQ ID NO: 45, and CDR3 comprising the amino acid sequence of SEQ ID NO: 46; or (f) a light chain variable region comprising CDR1 comprising the amino acid sequence of SEQ ID NO: 49, CDR2 comprising the amino acid sequence of SEQ ID NO: 50, and CDR3 comprising the amino acid sequence of SEQ ID NO: 51.The present invention provides an anti-uPAR antibody or an antigen-binding fragment thereof, comprising a heavy chain variable region and a light chain variable region comprising CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 52, CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 53, and CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 54.

[0251] A22. The anti-uPAR antibody or its antigen-binding fragment of A21 described above, wherein the heavy chain variable region comprises CDR1 having the amino acid sequence set forth in SEQ ID NO: 7, CDR2 having the amino acid sequence set forth in SEQ ID NO: 8, and CDR3 having the amino acid sequence set forth in SEQ ID NO: 9, and the light chain variable region comprises CDR1 having the amino acid sequence set forth in SEQ ID NO: 10, CDR2 having the amino acid sequence set forth in SEQ ID NO: 11, and CDR3 having the amino acid sequence set forth in SEQ ID NO: 12.

[0252] A23. Any one of the antibodies or antigen-binding fragments thereof among the above-mentioned A to A22, wherein the antibody or antigen-binding fragment thereof binds to uPAR or a fragment thereof comprising the amino acid sequence set forth in SEQ ID NO:61.

[0253] A24. The antibody sequence is in the light-heavy variable chain orientation (V L -V H ) any one of the antibodies A to A23 or antigen-binding fragments thereof.

[0254] A25. Any one of the antibodies A to A24 above, or an antigen-binding fragment thereof, wherein the antibody comprises a human variable region framework region.

[0255] A26. Any one of the antibodies or antigen-binding fragments thereof among the above-mentioned A to A25, which is fully human or an antigen-binding fragment thereof.

[0256] A27. Any one of the antibodies or antigen-binding fragments thereof selected from the above-mentioned A to A26, which is a chimeric antibody or an antigen-binding fragment thereof.

[0257] A28. Any one of the antibodies or antigen-binding fragments thereof selected from the above-mentioned A to A27, which is a humanized antibody or an antigen-binding fragment thereof.

[0258] A29. Any one of the antibodies or antigen-binding fragments thereof among the above-mentioned A to A28, wherein the antigen-binding fragment is Fab, Fab', F(ab')2, a variable fragment (Fv), or a single-chain variable region (scFv).

[0259] A30. The antibody or antigen-binding fragment thereof according to A29 above, wherein the antigen-binding fragment is an scFv.

[0260] A31. In certain non-limiting embodiments, the subject matter disclosed herein provides an antibody or antigen-binding fragment thereof that cross-competes with an antibody or antigen-binding fragment thereof of any one of A-A30 for binding to uPAR.

[0261] A32. In certain non-limiting embodiments, the subject matter disclosed herein provides an antibody or antigen-binding fragment thereof that binds to the same epitope region on uPAR as any one of the antibodies or antigen-binding fragments thereof of A-A30.

[0262] B. In certain non-limiting embodiments, the presently disclosed subject matter provides a composition comprising an antibody or antigen-binding fragment thereof of any one of A-A32.

[0263] B1. The composition of B above, which is a pharmaceutical composition further comprising a pharma- ceutically acceptable carrier.

[0264] C. In certain non-limiting embodiments, the presently disclosed subject matter provides an immunoconjugate comprising an antibody, or antigen-binding fragment thereof, of any one of A-A32, linked to a therapeutic agent.

[0265] C1. The immunoconjugate of C above, wherein the therapeutic agent is a drug, a cytotoxin, or a radioisotope.

[0266] D. In certain non-limiting embodiments, the presently disclosed subject matter provides compositions comprising an immunoconjugate of C or C1.

[0267] D1. The composition of D above, which is a pharmaceutical composition further comprising a pharma- ceutically acceptable carrier.

[0268] E. In certain non-limiting embodiments, the presently disclosed subject matter provides multispecific molecules comprising any one of antibodies, or antigen-binding fragments thereof, of A-A32, linked to one or more functional moieties.

[0269] E1. A multispecific molecule as defined in E above, wherein one or more functional moieties have a binding specificity different from that of an antibody or antigen-binding fragment thereof.

[0270] F. In certain non-limiting embodiments, the presently disclosed subject matter provides compositions comprising E or E1 multispecific molecules.

[0271] F1. The composition of F above, which is a pharmaceutical composition further comprising a pharma- ceutically acceptable carrier.

[0272] G. In certain non-limiting embodiments, the presently disclosed subject matter provides a nucleic acid encoding an antibody or antigen-binding fragment thereof of any one of A-A32.

[0273] G1. In certain non-limiting embodiments, the presently disclosed subject matter provides a vector comprising a nucleic acid molecule of G.

[0274] H. In certain non-limiting embodiments, the presently disclosed subject matter provides a host cell comprising a G vector or a G1 nucleic acid.

[0275] I. In certain non-limiting embodiments, the subject matter disclosed herein provides a method for detecting uPAR in a cell, tissue, or blood sample comprising contacting the cell, tissue, or blood sample with an antibody or antigen-binding fragment thereof of any one of A-A32, where the antibody or antigen-binding fragment thereof comprises a detectable label, and determining the amount of labeled antibody or antigen-binding fragment thereof bound to the cell, tissue, or blood sample by measuring the amount of detectable label associated with the cell, tissue, or blood sample, where the amount of bound antibody or antigen-binding fragment thereof indicates the amount of uPAR in the cell, tissue, or blood sample.

[0276] J. In certain non-limiting embodiments, the subject matter disclosed herein provides a method of treating or alleviating a disease or disorder in a subject, the method comprising administering to the subject an antibody or antigen-binding fragment thereof of any one of A-A32, an immunoconjugate of C or C1, a multispecific molecule of E or E1, or a composition of any one of B, B1, D, D1, F, and F1.

[0277] J1. The method of J above, wherein the disease or disorder is selected from the group consisting of tumors, aging-related conditions, and age-related tissue loss.

[0278] J2. The method of J1 above, wherein the disease or disorder is an aging-related pathology.

[0279] J3. The method of J1 above, wherein the aging-related pathology is selected from the group consisting of pulmonary fibrosis, atherosclerosis, Alzheimer's disease, diabetes, osteoarthritis, liver fibrosis, chronic kidney disease, cardiac fibrosis, and Parkinson's disease.

[0280] J4. The method of J2 above, wherein the disease or disorder is a tumor.

[0281] J5. The method of J4 above, wherein the tumor is selected from the group consisting of breast cancer, endometrial cancer, ovarian cancer, colon cancer, rectal cancer, lung cancer, stomach cancer, prostate cancer, gastric cancer, kidney cancer, pancreatic cancer, blood cancer, cervical cancer, head and neck cancer, liver cancer, urothelial cancer, melanoma, and brain cancer.

[0282] J6. The method of J5 above, wherein the hematological cancer is selected from the group consisting of acute lymphoblastic leukemia (ALL), chronic lymphocytic leukemia (CLL), acute myeloid leukemia (AML), myelofibrosis, polycythemia vera, myelodysplastic syndrome, and erythroleukemia.

[0283] J7. Any one of the methods J4 to J6 above, wherein the tumor is cancer.

[0284] K. In certain non-limiting embodiments, the subject matter disclosed herein provides a method of increasing production of immune activating cytokines in response to tumor cells in a subject, the method comprising administering to the subject an antibody or antigen-binding fragment thereof of any one of A-A32, an immunoconjugate of C or C1, a multispecific molecule of E or E1, or a composition of any one of B, B1, D, D1, F, and F1.

[0285] K1. The method of claim K, wherein the immune stimulating cytokine is selected from the group consisting of granulocyte-macrophage colony stimulating factor (GM-CSF), IFN-α, IFN-β, IFN-γ, TNF-α, IL-1, IL-2, IL-3, IL-6, IL-11, IL-7, IL-8, IL-12, IL-15, IL-21, interferon regulatory factor 7 (IRF7), CCL1, CCL2, CCL3, CCL5, CCL7, CCL8, CCL13, CCL16, CXCL1, CXCL3, CXCL5, CXCL9, CXCL10, and combinations thereof.

[0286] K2. Any one of the methods J-J7, K, and K1 above, wherein the subject is a human.

[0287] L. In certain non-limiting embodiments, the subject matter disclosed herein provides a kit for treating or alleviating a disease or disorder in a subject and / or increasing production of immune stimulatory cytokines in response to tumor cells in a subject, the kit comprising an antibody or antigen-binding fragment thereof of any one of A-A32, an immunoconjugate of C or C1, a multispecific molecule of E or E1, or a composition of any one of B, B1, D, D1, F, and F1.

[0288] L1. The kit of L above, wherein the kit further comprises written instructions for using the antibody or antigen-binding fragment thereof, immunoconjugate, multispecific molecule, or composition to treat or alleviate a disease or disorder in a subject and / or to increase production of immune stimulatory cytokines in response to tumor cells in a subject. EXAMPLES

[0289] The following examples are presented to provide those of skill in the art with a complete disclosure and description of how to make and use the antibodies, multispecific antibodies, compositions comprising them, screening, and treatment methods of the presently disclosed subject matter, and are not intended to limit the scope of what the inventors regard as the presently disclosed subject matter. It will be understood that various other embodiments may be practiced given the general description provided above.

[0290] Example 1 - Generation of anti-uPAR antibodies and scFv Recombinant uPAR from R&D (https: / / www.rndsystems.com / products / recombinant-human-upar-protein_807-uk) was used to generate the antibodies and antigen-binding fragments thereof disclosed herein. This recombinant uPAR is a human uPAR protein (Leu23-Arg303) with a C-terminal 6-his tag.

[0291] Six clones were generated: 8B1, 11E10, 17C9, 19D7, 6C8, and 14C5. The binding affinities K D The evaluation was carried out. D Values ​​were calculated using single cycle kinetic analysis in Biacore. K D is 9.52 x 10 -8 The K of 8B1 antibody was D is 3.48 x 10 -9 The K of the 17C9 antibody was D is 1.86 x 10 -8 The K of 6C8 antibody was D is 6.61 x 10 -9 The K of 14C5 antibody was D is 4.24 x 10 -8 It was M.

[0292] Embodiments of the subject matter disclosed herein From the above description, it will be apparent that variations and modifications may be made to the subject matter disclosed herein to adapt it to various applications and conditions. Such embodiments are also within the scope of the following claims.

[0293] The recitation of a list of elements in any definition of a variable herein includes definition of that variable as any single element or combination (or subcombination) of the listed elements. The recitation of an embodiment herein includes that embodiment as any single embodiment or in combination with any other embodiment or portion thereof.

[0294] All patents and publications mentioned in this specification are herein incorporated by reference to the same extent as if each individual patent and publication was specifically and individually indicated to be incorporated by reference.

Claims

1. An anti-uPAR antibody or an antigen-binding fragment thereof comprising a heavy chain variable region comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, at least about 100% homologous or identical to the amino acid sequence set forth in SEQ ID NO: 25, SEQ ID NO: 27, SEQ ID NO: 29, SEQ ID NO: 31, SEQ ID NO: 47, or SEQ ID NO:

55.

2. The anti-uPAR antibody or an antigen-binding fragment thereof according to claim 1, wherein the heavy chain variable region comprises an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, at least about 100% homologous or identical to the amino acid sequence set forth in SEQ ID NO:

27.

3. An anti-uPAR antibody or an antigen-binding fragment thereof comprising a light chain variable region comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, at least about 100% homologous or identical to the amino acid sequence set forth in SEQ ID NO: 26, SEQ ID NO: 28, SEQ ID NO: 30, SEQ ID NO: 32, SEQ ID NO: 48, or SEQ ID NO:

56.

4. The anti-uPAR antibody or an antigen-binding fragment thereof according to claim 3, wherein the light chain variable region comprises an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, at least about 100% homologous or identical to the amino acid sequence set forth in SEQ ID NO:

28.

5. (a) a heavy chain variable region comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or at least about 100% homologous or identical to the amino acid sequence set forth in SEQ ID NO: 25, SEQ ID NO: 27, SEQ ID NO: 29, SEQ ID NO: 31, SEQ ID NO: 47, or SEQ ID NO: 55, and (b) a light chain variable region comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or at least about 100% homologous or identical to the amino acid sequence set forth in SEQ ID NO: 26, SEQ ID NO: 28, SEQ ID NO: 30, SEQ ID NO: 32, SEQ ID NO: 48, or SEQ ID NO: 56, an anti-uPAR antibody or an antigen-binding fragment thereof. **Claim 6** An anti-uPAR antibody or an antigen-binding fragment thereof comprising a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region and the light chain variable region are (a) a heavy chain variable region comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or at least about 100% homologous or identical to the amino acid sequence set forth in SEQ ID NO: 25, and a light chain variable region comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or at least about 100% homologous or identical to the amino acid sequence set forth in SEQ ID NO: 26, (b) a heavy chain variable region comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, at least about 100% homologous or identical to the amino acid sequence set forth in SEQ ID NO: 27, and a light chain variable region comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, at least about 100% homologous or identical to the amino acid sequence set forth in SEQ ID NO: 28, (c) a heavy chain variable region comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, at least about 100% homologous or identical to the amino acid sequence set forth in SEQ ID NO: 29, and a light chain variable region comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, at least about 100% homologous or identical to the amino acid sequence set forth in SEQ ID NO: 30, (d) a heavy chain variable region comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, at least about 100% homologous or identical to the amino acid sequence set forth in SEQ ID NO: 31, and a light chain variable region comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, at least about 100% homologous or identical to the amino acid sequence set forth in SEQ ID NO: 32, (e) a heavy chain variable region comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, at least about 100% homologous or identical to the amino acid sequence set forth in SEQ ID NO: 47, and a light chain variable region comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, at least about 100% homologous or identical to the amino acid sequence set forth in SEQ ID NO: 48, and (f) an anti-uPAR antibody or an antigen-binding fragment thereof selected from the group consisting of a heavy chain variable region comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, at least about 100% homologous or identical to the amino acid sequence set forth in SEQ ID NO: 55, and a light chain variable region comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, at least about 100% homologous or identical to the amino acid sequence set forth in SEQ ID NO:

56.

7. The anti-uPAR antibody or an antigen-binding fragment thereof according to claim 6, wherein the heavy chain variable region comprises an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, at least about 100% homologous or identical to the amino acid sequence set forth in SEQ ID NO: 27, and the light chain variable region comprises an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, at least about 100% homologous or identical to the amino acid sequence set forth in SEQ ID NO:

28.

8. An anti-uPAR antibody or an antigen-binding fragment thereof, comprising a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 25, SEQ ID NO: 27, SEQ ID NO: 29, SEQ ID NO: 31, SEQ ID NO: 47, or SEQ ID NO:

55.

9. The anti-uPAR antibody or an antigen-binding fragment thereof according to claim 8, wherein the heavy chain variable region comprises the amino acid sequence set forth in SEQ ID NO:

27.

10. An anti-uPAR antibody or an antigen-binding fragment thereof, comprising a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 26, SEQ ID NO: 28, SEQ ID NO: 30, SEQ ID NO: 32, SEQ ID NO: 48, or SEQ ID NO:

56.

11. The anti-uPAR antibody or an antigen-binding fragment thereof according to claim 10, wherein the light chain variable region comprises the amino acid sequence set forth in SEQ ID NO:

28.

12. (a) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 25, SEQ ID NO: 27, SEQ ID NO: 29, SEQ ID NO: 31, SEQ ID NO: 47, or SEQ ID NO: 55, and (b) an anti-uPAR antibody or an antigen-binding fragment thereof, comprising a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 26, SEQ ID NO: 28, SEQ ID NO: 30, SEQ ID NO: 32, SEQ ID NO: 48, or SEQ ID NO:

56.

13. (a) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 25 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 26, (b) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 27 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 28, (c) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 29 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 30, (d) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 31 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 32, (e) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 47 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 48, and The antibody or antigen-binding fragment thereof according to claim 1, comprising a heavy-chain variable region containing the amino acid sequence set forth in SEQ ID NO: 55 and a light-chain variable region containing the amino acid sequence set forth in SEQ ID NO:

56.

14. The antibody or antigen-binding fragment thereof according to claim 13, wherein the heavy-chain variable region contains the amino acid sequence set forth in SEQ ID NO: 27 and the light-chain variable region contains the amino acid sequence set forth in SEQ ID NO:

28.

15. An anti-uPAR antibody or antigen-binding fragment thereof, comprising a heavy-chain variable region containing a CDR1 domain, a CDR2 domain, and a CDR3 domain, and a light-chain variable region containing a CDR1 domain, a CDR2 domain, and a CDR3 domain, wherein the CDR3 domains of the heavy-chain variable region and the light-chain variable region are (a) a heavy-chain variable region CDR3 containing the amino acid sequence set forth in SEQ ID NO: 3 and its conservative modifications, and a light-chain variable region CDR3 containing the amino acid sequence set forth in SEQ ID NO: 6 and its conservative modifications, (b) a heavy-chain variable region CDR3 containing the amino acid sequence set forth in SEQ ID NO: 9 and its conservative modifications, and a light-chain variable region CDR3 containing the amino acid sequence set forth in SEQ ID NO: 12 and its conservative modifications, (c) a heavy-chain variable region CDR3 containing the amino acid sequence set forth in SEQ ID NO: 15 and its conservative modifications, and a light-chain variable region CDR3 containing the amino acid sequence set forth in SEQ ID NO: 18 and its conservative modifications, (d) a heavy-chain variable region CDR3 containing the amino acid sequence set forth in SEQ ID NO: 21 and its conservative modifications, and a light-chain variable region CDR3 containing the amino acid sequence set forth in SEQ ID NO: 24 and its conservative modifications, (e) a heavy-chain variable region CDR3 containing the amino acid sequence set forth in SEQ ID NO: 43 and its conservative modifications, a light-chain variable region CDR3 containing the amino acid sequence set forth in SEQ ID NO: 46 and its conservative modifications, and An anti-uPAR antibody or an antigen-binding fragment thereof, selected from a heavy chain variable region CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 51 and its conservative modifications, and a light chain variable region CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 54 and its conservative modifications. **Claim 16** The CDR2 domains of the heavy chain variable region and the light chain variable region are (a) a heavy chain variable region CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 2 and its conservative modifications, and a light chain variable region CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 5 and its conservative modifications, (b) a heavy chain variable region CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 8 and its conservative modifications, and a light chain variable region CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 11 and its conservative modifications, (c) a heavy chain variable region CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 14 and its conservative modifications, and a light chain variable region CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 17 and its conservative modifications, (d) a heavy chain variable region CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 20 and its conservative modifications, and a light chain variable region CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 23 and its conservative modifications, (e) a heavy chain variable region CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 42 and its conservative modifications, a light chain variable region CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 45 and its conservative modifications, and (f) an antibody or an antigen-binding fragment thereof according to claim 15, selected from a heavy chain variable region CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 50 and its conservative modifications, and a light chain variable region CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 53 and its conservative modifications. **Claim 17** The CDR1 domains of the heavy chain variable region and the light chain variable region are (a) a heavy chain variable region CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 1 and its conservative modifications, and a light chain variable region CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 4 and its conservative modifications, (b) a heavy chain variable region CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 7 and its conservative modifications, and a light chain variable region CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 10 and its conservative modifications (c) a heavy chain variable region CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 13 and its conservative modifications, and a light chain variable region CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 16 and its conservative modifications (d) a heavy chain variable region CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 19 and its conservative modifications, and a light chain variable region CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 22 and its conservative modifications (e) a heavy chain variable region CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 41 and its conservative modifications, a light chain variable region CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 44 and its conservative modifications, and (f) The antibody or antigen-binding fragment thereof according to claim 15, selected from a heavy chain variable region CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 49 and its conservative modifications, and a light chain variable region CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 52 and its conservative modifications.

18. The antibody or antigen-binding fragment thereof according to claim 15, wherein one or more of the CDR sequences have up to about 5 amino acid substitutions.

19. The antibody or antigen-binding fragment thereof according to claim 15, wherein one or more of the CDR sequences have up to about 3 amino acid substitutions.

20. (a) CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 1, CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 2, and CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 3 (b) CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 7, CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 8, and CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 9 (c) CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 13, CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 14, and CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 15, (d) CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 19, CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 20, and CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 21, (e) CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 41, CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 42, and CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 43, or (f) an anti-uPAR antibody or an antigen-binding fragment thereof comprising a heavy chain variable region comprising CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 49, CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 50, and CDR3 comprising the amino acid sequence set forth in SEQ ID NO:

51.

21. (a) CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 4, CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 5, and CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 6, (b) CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 10, CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 11, and CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 12, (c) CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 16, CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 17, and CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 18, (d) CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 22, CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 23, and CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 24, (e) CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 44, CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 45, and CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46, or (f) an anti-uPAR antibody or an antigen-binding fragment thereof comprising a light chain variable region comprising CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 52, CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 53, and CDR3 comprising the amino acid sequence set forth in SEQ ID NO:

54.

22. (a) A heavy chain variable region comprising a CDR1 containing the amino acid sequence set forth in SEQ ID NO: 1, a CDR2 containing the amino acid sequence set forth in SEQ ID NO: 2, and a CDR3 containing the amino acid sequence set forth in SEQ ID NO: 3, and a CDR1 containing the amino acid sequence set forth in SEQ ID NO: 4, a CDR2 containing the amino acid sequence set forth in SEQ ID NO: 5, and a CDR3 containing the amino acid sequence set forth in SEQ ID NO: 6, which together form a light chain variable region. (b) A heavy chain variable region comprising a CDR1 containing the amino acid sequence set forth in SEQ ID NO: 7, a CDR2 containing the amino acid sequence set forth in SEQ ID NO: 8, and a CDR3 containing the amino acid sequence set forth in SEQ ID NO: 9, and a CDR1 containing the amino acid sequence set forth in SEQ ID NO: 10, a CDR2 containing the amino acid sequence set forth in SEQ ID NO: 11, and a CDR3 containing the amino acid sequence set forth in SEQ ID NO: 12, which together form a light chain variable region. (c) A heavy chain variable region comprising a CDR1 containing the amino acid sequence set forth in SEQ ID NO: 13, a CDR2 containing the amino acid sequence set forth in SEQ ID NO: 14, and a CDR3 containing the amino acid sequence set forth in SEQ ID NO: 15, and a CDR1 containing the amino acid sequence set forth in SEQ ID NO: 16, a CDR2 containing the amino acid sequence set forth in SEQ ID NO: 17, and a CDR3 containing the amino acid sequence set forth in SEQ ID NO: 18, which together form a light chain variable region. (d) A heavy chain variable region comprising a CDR1 containing the amino acid sequence set forth in SEQ ID NO: 19, a CDR2 containing the amino acid sequence set forth in SEQ ID NO: 20, and a CDR3 containing the amino acid sequence set forth in SEQ ID NO: 21, and a CDR1 containing the amino acid sequence set forth in SEQ ID NO: 22, a CDR2 containing the amino acid sequence set forth in SEQ ID NO: 23, and a CDR3 containing the amino acid sequence set forth in SEQ ID NO: 24, which together form a light chain variable region. (e) A heavy chain variable region comprising a CDR1 containing the amino acid sequence set forth in SEQ ID NO: 41, a CDR2 containing the amino acid sequence set forth in SEQ ID NO: 42, and a CDR3 containing the amino acid sequence set forth in SEQ ID NO: 43, and a CDR1 containing the amino acid sequence set forth in SEQ ID NO: 44, a CDR2 containing the amino acid sequence set forth in SEQ ID NO: 45, and a CDR3 containing the amino acid sequence set forth in SEQ ID NO: 46, which together form a light chain variable region, or (f) A heavy chain variable region comprising a CDR1 containing the amino acid sequence set forth in SEQ ID NO: 49, a CDR2 containing the amino acid sequence set forth in SEQ ID NO: 50, and a CDR3 containing the amino acid sequence set forth in SEQ ID NO: 51, and a CDR1 containing the amino acid sequence set forth in SEQ ID NO: 52, a CDR2 containing the amino acid sequence set forth in SEQ ID NO: 53, and a CDR3 containing the amino acid sequence set forth in SEQ ID NO: 54, an anti-uPAR antibody or an antigen-binding fragment thereof comprising a light chain variable region.

23. The anti-uPAR antibody or antigen-binding fragment thereof according to claim 22, wherein the heavy chain variable region comprises a CDR1 containing the amino acid sequence set forth in SEQ ID NO: 7, a CDR2 containing the amino acid sequence set forth in SEQ ID NO: 8, and a CDR3 containing the amino acid sequence set forth in SEQ ID NO: 9, and the light chain variable region comprises a CDR1 containing the amino acid sequence set forth in SEQ ID NO: 10, a CDR2 containing the amino acid sequence set forth in SEQ ID NO: 11, and a CDR3 containing the amino acid sequence set forth in SEQ ID NO:

12.

24. The antibody or antigen-binding fragment thereof according to claim 1, wherein the antibody or antigen-binding fragment thereof binds to uPAR or a fragment thereof containing the amino acid sequence set forth in SEQ ID NO:

61.

25. The antibody or antigen-binding fragment thereof according to claim 1, wherein the sequence of the antibody is in a light-heavy variable chain orientation (V Ｌ -V Ｈ ).

26. The antibody or antigen-binding fragment thereof according to claim 1, wherein the antibody comprises a human variable region framework region.

27. The antibody or antigen-binding fragment thereof according to claim 1, which is a fully human or antigen-binding fragment thereof.

28. The antibody or antigen-binding fragment thereof according to claim 1, which is a chimeric antibody or antigen-binding fragment thereof.

29. The antibody or antigen-binding fragment thereof according to claim 1, which is a humanized antibody or antigen-binding fragment thereof.

30. The antigen-binding fragment is Fab, Fab', F(ab') ２ , variable fragment (Fv), or single-chain variable region (scFv), the antibody or antigen-binding fragment according to claim 1.

31. The antigen-binding fragment is scFv, the antibody or antigen-binding fragment according to claim 30.

32. An antibody or its antigen-binding fragment that cross-competes with the antibody or its antigen-binding fragment according to any one of claims 1 to 31 for binding to uPAR.

33. An antibody or its antigen-binding fragment that binds to the same epitope region on uPAR as the antibody or its antigen-binding fragment according to any one of claims 1 to 31.

34. A composition comprising the antibody or its antigen-binding fragment according to claim 1.

35. The composition according to claim 34, which is a pharmaceutical composition further comprising a pharmaceutically acceptable carrier.

36. An immunoconjugate comprising the antibody or its antigen-binding fragment according to claim 1 conjugated to a therapeutic agent.

37. The immunoconjugate according to claim 36, wherein the therapeutic agent is a drug, a cytotoxin, or a radioisotope.

38. A composition comprising the immunoconjugate according to claim 36.

39. The composition according to claim 38, which is a pharmaceutical composition further comprising a pharmaceutically acceptable carrier.

40. A multispecific molecule comprising the antibody or its antigen-binding fragment according to claim 1 linked to one or more functional moieties.

41. The multispecific molecule according to claim 40, wherein the one or more functional moieties have a binding specificity different from that of the antibody or its antigen-binding fragment.

42. A composition comprising the multispecific molecule according to claim 40.

43. The composition according to claim 42, which is a pharmaceutical composition further comprising a pharmaceutically acceptable carrier.

44. A nucleic acid encoding the antibody according to claim 1 or an antigen-binding fragment thereof.

45. A vector comprising the nucleic acid molecule according to claim 44.

46. A host cell comprising the nucleic acid according to claim 44.

47. An in vitro or ex vivo method for detecting uPAR in a cell, tissue, or blood sample, comprising: contacting the cell, tissue, or blood sample with the antibody according to claim 1 or an antigen-binding fragment thereof, wherein the antibody or an antigen-binding fragment thereof comprises a detectable label; and determining the amount of the labeled antibody or an antigen-binding fragment thereof bound to the cell, tissue, or blood sample by measuring the amount of the detectable label associated with the cell, tissue, or blood sample, wherein the amount of the bound antibody or an antigen-binding fragment thereof indicates the amount of uPAR in the cell, tissue, or blood sample.

48. A composition for treating or alleviating a disease or disorder in a subject, comprising the composition comprising the antibody according to claim 1 or an antigen-binding fragment thereof.

49. The composition according to claim 48, wherein the disease or disorder is selected from the group consisting of tumors, age-related pathologies, and tissue involution associated with aging.

50. The composition according to claim 49, wherein the disease or disorder is an age-related pathology.

51. The composition according to claim 50, wherein the aging-related pathological condition is selected from the group consisting of pulmonary fibrosis, atherosclerosis, Alzheimer's disease, diabetes, osteoarthritis, liver fibrosis, chronic kidney disease, cardiac fibrosis, and Parkinson's disease.

52. The composition according to claim 48, wherein the disease or disorder is a tumor.

53. The composition according to claim 52, wherein the tumor is selected from the group consisting of breast cancer, endometrial cancer, ovarian cancer, colon cancer, rectal cancer, lung cancer, stomach cancer (stomach cancer), prostate cancer, stomach cancer (gastric cancer), kidney cancer, pancreatic cancer, blood cancer, cervical cancer, head and neck cancer, liver cancer, urothelial cancer, melanoma, and brain cancer.

54. The composition according to claim 53, wherein the blood cancer is selected from the group consisting of acute lymphoblastic leukemia (ALL), chronic lymphocytic leukemia (CLL), acute myeloid leukemia (AML), myelofibrosis, polycythemia vera, myelodysplastic syndrome, and erythroleukemia.

55. The composition according to claim 52, wherein the tumor is cancer.

56. A composition for increasing the production of an immune-activating cytokine in response to tumor cells in a subject, the composition comprising the antibody or antigen-binding fragment thereof according to claim 1.

57. The composition according to claim 56, wherein the immune-activating cytokine is selected from the group consisting of granulocyte macrophage colony-stimulating factor (GM-CSF), IFN-α, IFN-β, IFN-γ, TNF-α, IL-1, IL-2, IL-3, IL-6, IL-11, IL-7, IL-8, IL-12, IL-15, IL-21, interferon regulatory factor 7 (IRF7), CCL1, CCL2, CCL3, CCL5, CCL7, CCL8, CCL13, CCL16, CXCL1, CXCL3, CXCL5, CXCL9, CXCL10, and combinations thereof.

58. The composition according to claim 48, wherein the subject is a human. **Claim 59** A kit for treating or alleviating a disease or disorder in a subject and / or increasing the production of an immunostimulatory cytokine in response to tumor cells in the subject, the kit comprising the antibody or antigen-binding fragment thereof according to claim 1. **Claim 60** The kit according to claim 59, further comprising written instructions for using the antibody or antigen-binding fragment thereof, immunoconjugate, multispecific molecule, or composition for treating or alleviating a disease or disorder in a subject and / or increasing the production of an immunostimulatory cytokine in response to tumor cells in the subject.