Novel Class of Monospecific and Bispecific Humanized Antibodies that Target the Insulin-like Growth Factor Type I Receptor (IGF-1R)

Abstract

The present invention provides compositions and methods of use of anti-IGF-1R antibodies or antibody fragments. Preferably the antibodies bind to IGF-1R but not IR; are not agonists for IGF-1R; do not block binding of IGF-1 or IGF-2 to isolated IGF-1R, but effectively neutralize activation of IGF-1R by IGF-1 in intact cells; and block binding of an R1 antibody to IGF-1R. The antibodies may be murine, chimeric, humanized or human R1 antibodies comprising the heavy chain CDR sequences DYYMY (SEQ ID NO:1), YITNYGGSTYYPDTVKG (SEQ ID NO:2) and QSNYDYDGWFAY (SEQ ID NO:3) and the light chain CDR sequences KASQEVGTAVA (SEQ ID NO:4), WASTRHT (SEQ ID NO:5) and QQYSNYPLT (SEQ ID NO:6). Preferably the antibodies bind to an epitope of IGF-1R comprising the first half of the cysteine-rich domain of IGF-1R (residues 151-222). The anti-IGF-1R antibodies may be used for diagnosis or therapy of various diseases such as cancer.

What is claimed is: 
     
         1 . A method of treating cancer comprising administering to an individual with a cancer that expresses IGF-1R an anti-IGF-1R antibody or antigen binding fragment thereof, wherein said anti-IGF-1R antibody or fragment thereof comprises the heavy chain variable region complementarity determining region (CDR) sequences CDR1 (DYYMY, SEQ ID NO:1), CDR2 (YITNYGGSTYYPDTVKG, SEQ ID NO:2) and CDR3 (QSNYDYDGWFAY, SEQ ID NO:3) and the light chain variable region CDR sequences CDR1 (KASQEVGTAVA, SEQ ID NO:4), CDR2 (WASTRHT, SEQ ID NO:5) and CDR3 (QQYSNYPLT, SEQ ID NO:6). 
     
     
         2 . The method of  claim 1 , wherein the anti-IGF-1R antibody is a chimeric, humanized or human antibody. 
     
     
         3 . The method of  claim 1 , wherein the anti-IGF-1R antibody is a humanized R1 antibody comprising the amino acid sequences of SEQ ID NO:9 (hR1 VH) and SEQ ID NO:10 (hR1 VK). 
     
     
         4 . The method of  claim 1 , wherein said anti-IGF-1R antibody is a chimeric R1 (cR1) antibody comprising the amino acid sequences of SEQ ID NO:7 (R1 VH) and SEQ ID NO:8 (R1 VK) attached to human antibody constant region sequences. 
     
     
         5 . The method of  claim 1 , wherein said anti-IGF-1R antibody is a naked antibody. 
     
     
         6 . The method of  claim 5 , further comprising administering to the individual at least one therapeutic agent selected from the group consisting of an immunomodulator, an anti-angiogenic agent, a cytokine, a chemokine, a growth factor, a hormone, a drug, a prodrug, an enzyme, an oligonucleotide, an interference RNA, a pro-apoptotic agent, a photoactive therapeutic agent, a cytotoxic agent, a chemotherapeutic agent, a second antibody, a second antigen-binding antibody fragment and an immunoconjugate. 
     
     
         7 . The method of  claim 6 , wherein the at least one therapeutic agent is selected from the group consisting of an EGFR inhibitor, erlotinib, an anti-EGFR antibody, an IGF-1R inhibitor, a tryphostin, AG1024, AG538, a pyrrolo[2,3-d]-pyrimidine derivative, NVP-AEW541 and a second anti-IGF-1R antibody. 
     
     
         8 . The method of  claim 6 , wherein the at least one therapeutic agent is selected from the group consisting of 5-fluorouracil, aplidin, azaribine, anastrozole, anthracyclines, bendamustine, bleomycin, bortezomib, bryostatin-1, busulfan, calicheamycin, camptothecin, carboplatin, 10-hydroxycamptothecin, carmustine, celebrex, chlorambucil, cisplatin (CDDP), Cox-2 inhibitors, irinotecan (CPT-11), SN-38, carboplatin, cladribine, camptothecans, cyclophosphamide, cytarabine, dacarbazine, docetaxel, dactinomycin, daunorubicin, doxorubicin, 2-pyrrolinodoxorubicine (2P-DOX), cyano-morpholino doxorubicin, doxorubicin glucuronide, epirubicin glucuronide, estramustine, epidophyllotoxin, estrogen receptor binding agents, etoposide (VP16), etoposide glucuronide, etoposide phosphate, floxuridine (FUdR), 3′,5′-O-dioleoyl-FudR (FUdR-dO), fludarabine, flutamide, farnesyl-protein transferase inhibitors, gemcitabine, hydroxyurea, idarubicin, ifosfamide, L-asparaginase, lenolidamide, leucovorin, lomustine, mechlorethamine, melphalan, mercaptopurine, 6-mercaptopurine, methotrexate, mitoxantrone, mithramycin, mitomycin, mitotane, navelbine, nitrosurea, plicomycin, procarbazine, paclitaxel, pentostatin, PSI-341, raloxifene, semustine, streptozocin, tamoxifen, taxol, temazolomide, DTIC, transplatinum, thalidomide, thioguanine, thiotepa, teniposide, topotecan, uracil mustard, vinorelbine, vinblastine, vincristine and vinca alkaloids. 
     
     
         9 . The method of  claim 6 , wherein the immunomodulator is selected from the group consisting of IL-1, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-11, IL-12, IL-13, IL-14, IL-15, IL-16, IL-17, IL-18, IL-21, IL-25, interferon-alpha, interferon-beta, interferon-gamma, TNF-alpha and the stem cell growth factor designated “S1 factor. 
     
     
         10 . The method of  claim 1 , wherein said anti-IGF-1R antibody is attached to at least one therapeutic agent selected from the group consisting of a radionuclide, an immunomodulator, an anti-angiogenic agent, a cytokine, a chemokine, a growth factor, a hormone, a drug, a prodrug, an enzyme, an oligonucleotide, a pro-apoptotic agent, an interference RNA, a photoactive therapeutic agent, a cytotoxic agent, a chemotherapeutic agent, a second antibody, a second antigen-binding antibody fragment and a toxin. 
     
     
         11 . The method of  claim 10 , wherein the at least one therapeutic agent is selected from the group consisting of 5-fluorouracil, aplidin, azaribine, anastrozole, anthracyclines, bendamustine, bleomycin, bortezomib, bryostatin-1, busulfan, calicheamycin, camptothecin, carboplatin, 10-hydroxycamptothecin, carmustine, celebrex, chlorambucil, cisplatin (CDDP), Cox-2 inhibitors, irinotecan (CPT-11), SN-38, carboplatin, cladribine, camptothecans, cyclophosphamide, cytarabine, dacarbazine, docetaxel, dactinomycin, daunorubicin, doxorubicin, 2-pyrrolinodoxorubicine (2P-DOX), cyano-morpholino doxorubicin, doxorubicin glucuronide, epirubicin glucuronide, estramustine, epidophyllotoxin, estrogen receptor binding agents, etoposide (VP16), etoposide glucuronide, etoposide phosphate, floxuridine (FUdR), 3′,5′-O-dioleoyl-FudR (FUdR-dO), fludarabine, flutamide, farnesyl-protein transferase inhibitors, gemcitabine, hydroxyurea, idarubicin, ifosfamide, L-asparaginase, lenolidamide, leucovorin, lomustine, mechlorethamine, melphalan, mercaptopurine, 6-mercaptopurine, methotrexate, mitoxantrone, mithramycin, mitomycin, mitotane, navelbine, nitrosurea, plicornycin, procarbazine, paclitaxel, pentostatin, PSI-341, raloxifene, semustine, streptozocin, tamoxifen, taxol, temazolomide, DTIC, transplatinum, thalidomide, thioguanine, thiotepa, teniposide, topotecan, uracil mustard, vinorelbine, vinblastine, vincristine and vinca alkaloids. 
     
     
         12 . The method of  claim 10 , wherein the immunomodulator is selected from the group consisting of a cytokine, a stem cell growth factor, a lymphotoxin, an hematopoietic factor, a colony stimulating factor (CSF), an interferon (IFN), erythropoietin, thrombopoietin, granulocyte-colony stimulating factor (G-CSF), granulocyte macrophage-colony stimulating factor (GM-CSF), interferon-α, interferon-β, interferon-γ, S1 factor, human growth hormone, N-methionyl human growth hormone, bovine growth hormone, parathyroid hormone, thyroxine, insulin, proinsulin, relaxin, prorelaxin, follicle stimulating hormone (FSH), thyroid stimulating hormone (TSH), luteinizing hormone (LH), hepatic growth factor, prostaglandin, fibroblast growth factor, prolactin, placental lactogen, OB protein, tumor necrosis factor-α, tumor necrosis factor-β, mullerian-inhibiting substance, mouse gonadotropin-associated peptide, inhibin, activin, vascular endothelial growth factor, integrin, NGF-β, platelet-growth factor, TGF-α, TGF-β, insulin-like growth factor-I, insulin-like growth factor-II, macrophage-CSF, IL-1, IL-1α, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-11 IL-12, IL-13, IL-14, IL-15, IL-16, IL-17, IL-18, IL-21, IL-25, LIF, kit-ligand, angiostatin, thrombospondin, endostatin, and lymphotoxin. 
     
     
         13 . The method of  claim 10 , wherein the toxin is selected from the group consisting of ricin, abrin, alpha toxin, saporin, ribonuclease (RNase), e.g., onconase, DNase I, Staphylococcal enterotoxin-A, pokeweed antiviral protein, gelonin, diphtheria toxin,  Pseudomonas  exotoxin, and  Pseudomonas  endotoxin. 
     
     
         14 . The method of  claim 10 , wherein the radionuclide is selected from the group consisting of  111 In,  177 Lu,  212 Bi,  213 Bi,  211 At,  62 Cu,  67 Cu,  90 Y,  125 I,  131 I,  32 P,  33 P,  47 Sc,  111 Ag,  67 Ga,  142 Pr,  153 Sm,  161 Tb,  166 Dy,  166 Ho,  186 Re,  188 Re,  189 Re,  212 Pb,  223 Ra,  225 Ac,  59 Fe,  75 Se,  77 As,  89 Sr,  99 Mo,  105 Rh,  109 Pd,  143 Pr,  149 Pm,  169 Er,  194 Ir,  198 Au,  199 Au,  211 Pb,  58 Co,  67 Ga,  80m Br,  99m Tc,  103m Rh,  109 Pt,  111 In,  119 Sb,  161 Ho,  189m Os,  192 Ir,  152 Dy,  211 At,  212 Bi,  223 Ra,  219 Rn,  215 Po,  211 Bi,  225 Ac,  221 Fr,  217 At,  225 Fm,  11 C,  13 N,  15 O,  75 Br,  198 Au,  224 Ac,  126 I,  133 I,  77 Br,  113m In,  95 Ru,  97 Ru,  103 Ru,  105 Ru,  107 Hg,  203 Hg,  121m Te,  122m Te,  125m Te,  165 Tm,  167 Tm,  168 Tm,  197 Pt,  109 Pd,  105 Rh,  142 Pr,  143 Pr,  161 Tb,  166 Ho,  199 Au,  57 Co,  58 Co,  51 Cr,  59 Fe,  75 Se,  201 Tl,  76 Br,  169 Yb, and an alpha-emitter. 
     
     
         15 . The method of  claim 1 , wherein said anti-IGF-1R antibody or antigen binding fragment thereof comprises constant region sequences of a human IgG1 or IgG4 antibody. 
     
     
         16 . The method of  claim 1 , wherein the cancer is selected from the group consisting of Wilms' tumor, Ewing sarcoma, neuroblastoma, neuroendocrine tumors, melanoma, glioblastoma, breast, colon, rectal, gastric, prostate, liver, renal, biliary, pancreatic, lung, endometrial, cervical, ovarian, esophageal, medullary thyroid, bladder, head-and-neck, skin cancer, acute lymphoblastic leukemia, acute myelogenous leukemia, chronic lymphocytic leukemia, chronic myelogenous leukemia, Hodgkin's lymphoma, non-Hodgkin's lymphoma, multiple myeloma, astrocytoma and glioma. 
     
     
         17 . The method of  claim 1 , wherein said anti-IGF-1R antibody or fragment thereof is part of a fusion protein or a bispecific antibody. 
     
     
         18 . The method of  claim 17 , wherein the bispecific antibody comprises a second antibody or antigen binding fragment thereof that binds to a tumor-associated antigen. 
     
     
         19 . The method of  claim 18 , wherein the tumor-associated antigen is selected from the group consisting of carbonic anhydrase IX, CCCL19, CCCL21, CSAp, CD1, CD1a, CD2, CD3, CD4, CD5, CD8, CD11A, CD14, CD15, CD16, CD18, CD19, CD20, IGF-1R, CD21, CD22, CD23, CD25, CD29, CD30, CD32b, CD1, CD37, CD38, CD40, CD40L, CD45, CD46, CD52, CD54, CD55, CD59, CD64, CD66a-e, CD67, CD70, CD74, CD79a, CD80, CD83, CD95, CD126, CD11, CD138, CD147, CD154, CEACAM5, CEACAM6, B7, ED-B fibronectin, Factor H, FHL-1, Flt-3, folate receptor, GROB, HMGB-1, hypoxia inducible factor (HIF), HM1.24, insulin-like growth factor-1 (ILGF-1), IFN-γ, IFN-α, IFN-β, IL-2, IL-4R, IL-6R, IL-13R, IL-15R, IL-17R, IL-18R, IL-6, IL-8, IL-12, IL-15, IL-17, IL-18, IL-25, IP-10, MAGE, mCRP, MCP-1, MIP-1A, MIP-1B, MIF, MUC1, MUC2, MUC3, MUC4, MUC5, PAM4 antigen, NCA-95, NCA-90, PSMA, EGP-1, EGP-2, AFP, Ia, HM1.24, HLA-DR, tenascin, Le(y), RANTES, T101, TAC, Tn antigen, Thomson-Friedenreich antigens, tumor necrosis antigens, TNF-α, TRAIL receptor (R1 and R2), VEGFR, EGFR, PlGF, complement factors C3, C3a, C3b, C5a, C5, and an oncogene product. 
     
     
         20 . The method of  claim 18 , wherein the second antibody is selected from the group consisting of the hPAM4, hA20, hA19, hIMMU31, hLL1, hLL2, hMu-9, hL243, hMN-14, hMN-15, hMN-3, hRS7, h679 and h734. 
     
     
         21 . The method of  claim 17 , wherein the bispecific antibody comprises a second antibody or antigen-binding fragment thereof that binds to a hapten on a targetable construct. 
     
     
         22 . The method of  claim 21 , further comprising administering to the individual a targetable construct comprising at least one hapten that binds to the second antibody or fragment thereof, wherein the targetable construct comprises at least one therapeutic agent.