Humanized L243 Antibodies

Abstract

Humanized antibodies are provided that specifically bind HLA-DR. The antibodies recognize the epitope recognized by the murine monoclonal antibody L243. Processes for preparing such antibodies, pharmaceutical compositions containing such antibodies, and clinical therapeutic and diagnostic, as well as research-related uses for such antibodies, are provided.

What is claimed is: 
     
         1 . A method of treating multiple myeloma comprising administering a humanized anti-HLA-DR antibody or fragment thereof to a human subject with an HLA-DR positive multiple myeloma. 
     
     
         2 . A method of treating acute lymphocytic leukemia comprising administering a humanized anti-HLA-DR antibody or fragment thereof to a human subject with an HLA-DR positive acute lymphocytic leukemia (ALL). 
     
     
         3 . A method of treating chronic lymphocytic leukemia comprising administering a humanized anti-HLA-DR antibody or fragment thereof to a human subject with an HLA-DR positive chronic lymphocytic leukemia (CLL). 
     
     
         4 . A method of treating hairy cell leukemia comprising administering a humanized anti-HLA-DR antibody or fragment thereof to a human subject with an HLA-DR positive hairy cell leukemia. 
     
     
         5 . A method of treating a B-cell lymphoma or leukemia comprising subcutaneously administering an anti-HLA-DR antibody or fragment thereof to a human subject with a B-cell lymphoma or leukemia. 
     
     
         6 . The method of  claim 1 , wherein the humanized anti-HLA-DR antibody or fragment thereof comprises human IgG4 constant region sequences and wherein the humanized anti-HLA-DR antibody or fragment thereof comprises a Ser241Pro point mutation in the hinge region of the antibody or fragment thereof. 
     
     
         7 . The method of  claim 1 , wherein the humanized anti-HLA-DR antibody or antigen-binding fragment thereof is a naked antibody or fragment thereof. 
     
     
         8 . The method of  claim 7 , further comprising administering at least one therapeutic agent to the subject. 
     
     
         9 . The method of  claim 8 , wherein the therapeutic agent is selected from the group consisting of antibodies, antibody fragments, drugs, chemotherapeutic agents, toxins, hormones, hormone antagonists, immunomodulators and cytokines. 
     
     
         10 . The method of  claim 9 , wherein the chemotherapeutic agent is a taxane, a nitrogen mustard, an ethylenimine, an alkyl sulfonate, a nitrosourea, a triazene, a folic acid analog, a pyrimidine analog, a purine analog, an antibiotic, a platinum coordination complex, a COX-2 inhibitor, an apoptotic agent, a substituted urea, a methyl hydrazine, a steroid, a progestin, an estrogen, an antiestrogen, an androgen, a camptothecin, actinomycin, azaribine, anastrozole, azacytidine, bleomycin, bryostatin-1, busulfan, carmustine, celecoxib, chlorambucil, cisplatinum, irinotecan, carboplatin, cladribine, cyclophosphamide, cytarabine, dacarbazine, docetaxel, dacarbazine, dactinomycin, daunorubicin, dexamethasone, diethylstilbestrol, doxorubicin, ethinyl estradiol, estramustine, etoposide, floxuridine, fludarabine, flutamide, 5-fluorouracil, fluoxymesterone, gemcitabine, hydroxyprogesterone caproate, hydroxyurea, idarubicin, ifosfamide, L-asparaginase, leucovorin, lomustine, mechlorethamine, medroprogesterone acetate, megestrol acetate, melphalan, mercaptopurine, methotrexate, mitoxantrone, mitomycin, mitotane, oxaliplatin, phenyl butyrate, prednisone, procarbazine, paclitaxel, pentostatin, semustine, streptozocin, SN-38, tamoxifen, taxanes, testosterone propionate, thalidomide, thioguanine, teniposide, topotecan, uracil mustard, vinblastine, vinorelbine or vincristine. 
     
     
         11 . The method of  claim 9 , wherein the therapeutic agent is a second antibody or antigen-binding fragment thereof that binds to a tumor-associated antigen. 
     
     
         12 . The method of  claim 11 , wherein the tumor-associated antigen is selected from the group consisting of A3, BrE3-antigen, CD1, CD1a, CD3, CD5, CD15, CD19, CD20, CD21, CD22, CD23, CD25, CD30, CD45, CD74, CD79a, CD80, HLA-DR, NCA95, NCA90, HCG, CEA (CEACAM-5), CEACAM-6, CSAp, EGFR, EGP-1, EGP-2, Ep-CAM, Ba 733, HER2/neu, hypoxia inducible factor (HIF), KC4-antigen, KS-1 antigen, KS1-4, Le-Y, macrophage inhibition factor (MIF), MAGE, MUC1, MUC2, MUC3, MUC4, MUC16, PAM-4-antigen, PSA, PSMA, RS5, S100, TAG-72, p53, tenascin, IL-6, IL-8, insulin growth factor-1 (IGF-1), Tn antigen, tumor necrosis antigens, VEGF, 17-1A-antigen, an angiogenesis marker, ED-B fibronectin, an oncogene marker, an oncogene product, HM1.24, VEGF, ILGF, placental growth factor and carbonic anhydrase IX. 
     
     
         13 . The method of  claim 9 , wherein the immunomodulator is selected from the group consisting of a cytokine, a stem cell growth factor, a lymphotoxin, a tumor necrosis factor (TNF), TNF-α, a hematopoietic factor, an interleukin, IL-1, IL-2, IL-3, IL-6, IL-10, IL-12, IL-18, IL-21, a colony stimulating factor, G-CSF, GM-CSF, interferon-α, -β or -γ, erythropoietin and thrombopoietin. 
     
     
         14 . The method of  claim 1 , wherein the humanized anti-HLA-DR antibody or antigen-binding fragment thereof is conjugated to at least one therapeutic or diagnostic agent. 
     
     
         15 . The method of  claim 14 , wherein the therapeutic agent is selected from the group consisting of antibodies, antibody fragments, drugs, chemotherapeutic agents, toxins, enzymes, nucleases, hormones, hormone antagonists, immunomodulators, cytokines, chelators, boron compounds, photoactive agents, dyes and radioisotopes. 
     
     
         16 . The method of  claim 1 , wherein the humanized anti-HLA-DR antibody or antigen-binding fragment thereof is administered intravenously, subcutaneously, or intramuscularly at a dose of between 20 and 2000 mg. 
     
     
         17 . The method  claim 1 , wherein the humanized anti-HLA-DR antibody or antigen-binding fragment thereof is conjugated to one or more lipids, polymeric carriers, micelles, nanoparticles, or a combination thereof. 
     
     
         18 . The method of  claim 15 , wherein the chemotherapeutic agent is selected from the group consisting of a taxane, a nitrogen mustard, an ethylenimine, an alkyl sulfonate, a nitrosourea, a triazene, a folic acid analog, a pyrimidine analog, a purine analog, an antibiotic, a platinum coordination complex, a COX-2 inhibitor, an apoptotic agent, a substituted urea, a methyl hydrazine, a steroid, a progestin, an estrogen, an antiestrogen, an androgen, a camptothecin, actinomycin, azaribine, anastrozole, azacytidine, bleomycin, bryostatin-1, busulfan, carmustine, celecoxib, chlorambucil, cisplatinum, irinotecan (CPT-11), carboplatinum, cladribine, cyclophosphamide, cytarabine, dacarbazine, docetaxel, dacarbazine, dactinomycin, daunorubicin, dexamethasone, diethylstilbestrol, doxorubicin, ethinyl estradiol, estramustine, etoposide, floxuridine, fludarabine, flutamide, 5-fluorouracil, fluoxymesterone, gemcitabine, hydroxyprogesterone caproate, hydroxyurea, idarubicin, ifosfamide, L-asparaginase, leucovorin, lomustine, mechlorethamine, medroprogesterone acetate, megestrol acetate, melphalan, mercaptopurine, methotrexate, mitoxantrone, mitomycin, mitotane, oxaliplatin, phenyl butyrate, prednisone, procarbazine, paclitaxel, pentostatin, semustine, streptozocin, SN-38, tamoxifen, taxanes, testosterone propionate, thalidomide, thioguanine, teniposide, topotecan, uracil mustard, vinblastine, vinorelbine and vincristine. 
     
     
         19 . The method of  claim 15 , wherein the toxin is selected from the group consisting of ricin, abrin, ribonuclease, DNase I, Staphylococcal enterotoxin-A, pokeweed antiviral protein, gelonin, diphtheria toxin, Pseudomonas exotoxin and Pseudomonas endotoxin. 
     
     
         20 . The method of  claim 15 , wherein the immunomodulator is selected from the group consisting of a cytokine, a stem cell growth factor, a lymphotoxin, a tumor necrosis factor (TNF), TNF-α, a hematopoietic factor, an interleukin, IL-1, IL-2, IL-3, IL-6, IL-10, IL-12, IL-18, IL-21, a colony stimulating factor, G-CSF, GM-CSF, interferon-α, -β or -γ, erythropoietin and thrombopoietin. 
     
     
         21 . The method of  claim 15 , wherein the cytokine is interferon-α, interferon-β, interferon-γ or GM-CSF. 
     
     
         22 . The method of  claim 15 , wherein the radioisotope is selected from the group consisting of In-111, Lu-177, Bi-212, Bi-213, At-211, Cu-62, Cu-64, Cu-67, Y-90, 1-125, 1-131, P-32, P-33, Sc-47, Ag-111, Ga-67, Pr-142, Sm-153, Tb-161, Dy-166, Ho-166, Re-186, Re-188, Re-189, Pb-212, Ra-223, Ac-225, Fe-59, Se-75, As-77, Sr-89, Mo-99, Rh-105, Pd-109, Pr-143, Pm-149, Er-169, Ir-194, Au-198, Au-199, Ac-225 and Pb-211. 
     
     
         23 . The method of  claim 15 , wherein the radioisotope is an alpha-particle-emitting radionuclide. 
     
     
         24 . The method of  claim 23 , wherein the alpha particle has a decay energy of between 2,000 and 10,000 keV. 
     
     
         25 . The method of  claim 23 , wherein the alpha particle has a decay energy of between 3,000 and 8,000 keV. 
     
     
         26 . The method of  claim 23 , wherein the alpha particle has a decay energy of between 4,000 and 7,000 keV. 
     
     
         27 . The method of  claim 1 , wherein the humanized anti-HLA-DR antibody is a humanized L243 antibody comprising heavy chain variable domain complementarity determining region (CDR) sequences CDR1 (NYGMN, residues 31 to 35 of SEQ ID NO: 4), CDR2 (WINTYTREPTYADDFKG, residues 50 to 66 of SEQ ID NO:4), and CDR3 (DITAVVPTGFDY, residues 99 to 110 of SEQ ID NO:4) and heavy chain framework residues F27, K38, K46, A68 and F91 and light chain variable domain CDR sequences CDR1 (RASENIYSNLA, residues 24 to 34 of SEQ ID NO:2), CDR2 (AASNLAD, residues 50 to 56 of SEQ ID NO:2), and CDR3 (QHFWTTPWA, residues 89 to 98 of SEQ ID NO:2) and light chain framework residues R37, K39, V48 and F49, wherein the remainder of the humanized anti-HLA-DR antibody framework region and constant region sequences are from one or more human antibodies. 
     
     
         28 . The method of  claim 1 , wherein the humanized anti-HLA-DR antibody or fragment thereof comprises the hL243VK amino acid sequence SEQ ID NO:6 and the hL243VH amino acid sequence SEQ ID NO:8. 
     
     
         29 . A method of treating Hodgkins or non-Hodgkin's lymphoma comprising administering a humanized anti-HLA-DR antibody or fragment thereof to a human subject with Hodgkins or non-Hodgkin's lymphoma, wherein the antibody or fragment is administered subcutaneously. 
     
     
         30 . The method of  claim 29 , wherein the antibody or fragment thereof is administered at a dosage of 200 to 800 mg weekly. 
     
     
         31 . The method of  claim 29 , wherein administration is repeated for at least 2 weeks. 
     
     
         32 . The method of  claim 29 , wherein administration is repeated for between 2 to 12 weeks. 
     
     
         33 . The method of  claim 31 , further comprising administering a maintenance dosage of the antibody or fragment thereof for 1 or 2 weeks every 2 to 3 months.