TARGETED BINDING AGENTS DIRECTED TO UPAR AND USES THEREOF
Targeted binding agents directed to the antigen uPAR and uses of such antibodies are described. In particular, fully human monoclonal antibodies directed to the antigen uPAR. Nucleotide sequences encoding, and amino acid sequences comprising, heavy and light chain immunoglobulin molecules, particularly sequences corresponding to contiguous heavy and light chain sequences spanning the framework regions and/or complementarity determining regions (CDR's), specifically from FR1 through FR4 or CDR1 through CDR3. Hybridomas or other cell lines expressing such immunoglobulin molecules and monoclonal antibodies.
1 - 63 . (canceled)
64 . An isolated targeted binding agent that specifically binds to urokinase-type plasminogen activator receptor (uPAR) with a Kd of less than 10 nanomolar and inhibits binding of urokinase-type plasminogen activator (uPA) to uPAR.
65 . The targeted binding agent of claim 64 , wherein said targeted binding agent at a concentration of 1 μg/ml inhibits greater than 90% of the activation of plasminogen by 6×10 4 U937 cells spiked with 3 nM scuPA.
66 . The targeted binding agent of claim 64 , wherein said targeted binding agent at a concentration of 1 μg/ml inhibits the activation of plasminogen by 6×10 4 U937 cells spiked with 3 nM scuPA with an IC 50 of less than 0.08 μg/ml.
67 . The targeted binding agent of claim 64 , wherein said targeted binding agent at a concentration of 2 μg/ml incubated with 18×10 4 U937 cells in 100 ul inhibits greater than 80% of uPAR-mediated cell adhesion of U937 cells spiked with 25 nM scuPA to tissue culture plastic coated with 5 □g/ml vitronectin.
68 . The targeted binding agent of claim 64 , wherein said targeted binding agent at a concentration of 10 μg/ml incubated with 10 4 HT1080 cells inhibits greater than 90% of cell invasion through a gel comprising 7.7 mg per ml Matrigel™.
69 . The targeted binding agent of claim 64 , wherein said targeted binding agent inhibits tumor growth and metastasis in a mammal.
70 . The targeted binding agent of claim 64 , wherein said targeted binding agent binds uPAR with a Kd of less than 500 picomolar (pM).
71 . A targeted binding agent according to claim 64 , wherein said targeted binding agent is
any one of the monoclonal antibodies as shown in Table 1;
monoclonal antibody 2.19.2 (ATCC Accession Number PTA-7474);
monoclonal antibody 2.6.1 (ATCC Accession Number PTA-7475); or
monoclonal antibody 4.18.1 (ATCC Accession Number PTA-7476).
72 . A targeted binding agent wherein said targeted binding agent is derivable from the monoclonal antibody of claim 71 .
73 . The targeted binding agent according to claim 64 , wherein said targeted binding agent comprises a polypeptide comprising the sequence of
SEQ ID NO.: 26, wherein SEQ ID NO.: 26 has any one of the unique combinations of germline and non-germline residues indicated by each row of Table 15;
SEQ ID NO.: 28, wherein SEQ ID NO.: 28 has any one of the unique combinations of germline and non-germline residues indicated by each row of Table 14;
SEQ ID NO.: 30, wherein SEQ ID NO.: 30 has any one of the unique combinations of germline and non-germline residues indicated by each row of Table 13;
SEQ ID NO.: 32, wherein SEQ ID NO.: 32 has any one of the unique combinations of germline and non-germline residues indicated by each row of Table 12;
SEQ ID NO.: 50, wherein SEQ ID NO.: 50 has any one of the unique combinations of germline and non-germline residues indicated by each row of Table 17; or
SEQ ID NO.: 52, wherein SEQ ID NO.: 52 has any one of the unique combinations of germline and non-germline residues indicated by each row of Table 16.
74 . The targeted binding agent according to claim 64 , wherein said targeted binding agent comprises a polypeptide having the sequence of
SEQ ID NO.: 26;
SEQ ID NO.: 28;
SEQ ID NO.: 30;
SEQ ID NO.: 32;
SEQ ID NO.: 50; or
SEQ ID NO.: 52.
75 . A targeted binding agent which cross-competes with the targeted binding agent of claim 64 for binding to uPAR.
76 . A targeted binding agent that binds to the same epitope on uPAR as the targeted binding agent of claim 64 .
77 . A targeted binding agent comprising an amino acid sequence, comprising
a) any one of a CDR1, CDR2 or CDR3 sequence as shown in Table 18 or Table 19;
b) a CDR1, CDR2 and CDR3 sequence as shown in Table 18;
c) a CDR1, CDR2 and CDR3 sequence as shown in Table 19; or
d) a CDR1, CDR2 and CDR3 sequence as shown in Table 18 and a CDR1, CDR2 and CDR3 sequence as shown in Table 19.
78 . A targeted binding agent according to claim 77 wherein said targeted binding agent is:
an antibody;
a monoclonal antibody;
a fully human monoclonal antibody; or
a binding fragment of a fully human monoclonal antibody.
79 . The targeted binding agent of claim 78 , wherein said binding fragment is selected from the group consisting of Fab, Fab′, F(ab′) 2 , Fv and Dab.
80 . A nucleic acid molecule encoding the targeted binding agent of claim 77 .
81 . A vector comprising the nucleic acid molecule of claim 80 .
82 . A host cell comprising the vector of claim 81 .
83 . A method of treating a malignant tumor in an animal, comprising:
selecting an animal in need of treatment for a malignant tumor; and
administering to said animal a therapeutically effective dose of the targeted binding agent of claim 64 .
84 . The method of claim 83 , wherein said malignant tumor is selected from the group consisting of: melanoma, small cell lung cancer, non-small cell lung cancer, glioma, hepatocellular (liver) carcinoma, thyroid tumor, gastric (stomach) cancer, prostate cancer, breast cancer, ovarian cancer, bladder cancer, lung cancer, glioblastoma, endometrial cancer, kidney cancer, colon cancer, pancreatic cancer, esophageal carcinoma, head and neck cancers, mesothelioma, sarcomas, biliary (cholangiocarcinoma), small bowel adenocarcinoma, pediatric malignancies and epidermoid carcinoma.
85 . A method of treating uPAR induced disease-related cell adhesion and/or invasion, comprising:
selecting an animal in need of treatment for disease-related cell adhesion and/or invasion; and
administering to said animal a therapeutically effective dose of the targeted binding agent of claim 64 .
86 . The method of claim 85 , wherein said disease-related cell adhesion and/or invasion is tumor metastasis.
87 . A method of treating a non-neoplastic disease, comprising:
selecting an animal in need of treatment for a non-neoplastic disease; and
administering to said animal a therapeutically effective dose of the targeted binding agent of claim 64 .
88 . The method of claim 87 , wherein the non-neoplastic disease is selected from the group consisting of arthritis, atherosclerosis and allergic conjunctivitis.
89 . A method of producing an anti-uPAR antibody of the invention by culturing the host cell of claim 82 under conditions wherein a nucleic acid molecule is expressed to produce the antibody, followed by recovering the antibody.