Antibody selective for dr4 and uses thereof

Abstract

An antibody of the invention interacts with human DR5 or with human DR4 to produce agonistic or antagonistic effects downstream of the receptor including inhibition of cell proliferation and apoptosis. Nucleic acid sequences and amino acid sequences of DR5 and DR4 antibodies have been elucidated and vectors and cells containing and expressing these sequences have been generated. Methods and uses for the antibodies are detailed including treatment of apoptosis-related disease and treatment of dysregulated cell growth.

1 . A purified antibody which specifically binds a TRAIL receptor DR4 and does not bind TRAIL receptor DR5, DcR1 or DcR2, wherein the antibody induces apoptosis in vitro in at least 50% of target cells expressing DR4 at an antibody concentration of less than 100 ng/ml. 
     
     
         2 . The purified antibody of  claim 1 , wherein said antibody is a monoclonal antibody. 
     
     
         3 . The purified antibody of  claim 1 , wherein the DR4 is human DR4. 
     
     
         4 . The purified antibody of  claim 1 , wherein the target cell is a cancer cell. 
     
     
         5 . The purified antibody of  claim 1 , wherein the target cell is a rheumatoid arthritis synovial cell. 
     
     
         6 . The purified antibody of  claim 1 , wherein the target cell is a lymphocyte. 
     
     
         7 . The antibody of  claim 1 , wherein said antibody is humanized. 
     
     
         8 . A method of selectively inducing apoptosis or inhibiting proliferation in target cells expressing DR4, comprising the step of contacting the target cells with a therapeutic quantity of the antibody of  claim 1 . 
     
     
         9 . The method of  claim 8 , wherein the contacting is in vivo. 
     
     
         10 . The method of  claim 8 , wherein the contacting is in vitro. 
     
     
         11 . The method of  claim 8 , wherein the target cell is a cancer cell. 
     
     
         12 . The method of  claim 8 , wherein the target cell is a rheumatoid arthritis synovial cell. 
     
     
         13 . The method of  claim 8 , wherein the target cell is a lymphocyte. 
     
     
         14 . The method of  claim 8 , wherein the target cell is a virally infected cell. 
     
     
         15 . The method of  claim 8 , further comprising contacting the target cells with a therapeutic quantity of an antibody which specifically binds a TRAIL receptor DR5, wherein said second antibody, in its soluble form, has in vivo and in vitro apoptosis-inducing activity in target cells expressing DR5. 
     
     
         16 . The method of  claim 8 , further comprising contacting the target cells with a therapeutic agent. 
     
     
         17 . The method of  claim 16 , wherein the therapeutic agent is a chemotherapeutic agent. 
     
     
         18 . The method of  claim 16 , wherein the therapeutic agent is selected from the group consisting of bleomycin, carboplatin, chlorambucil, cisplatin, colchicine, cyclophosphamide, daunorubicin, actinomycin, diethylstilbestrol doxorubicin, etoposide, 5-fluorouracil, floxuridine, melphalan, methotrexate, mitomycin, 6-mercaptopurine, teniposide, 6-thioguanine, vincristine and vinblastine. 
     
     
         19 . The method of  claim 16 , wherein the therapeutic agent is doxorubicin. 
     
     
         20 . The method of  claim 16 , wherein the therapeutic agent is paclitaxel. 
     
     
         21 . The method of  claim 16 , wherein the therapeutic agent is methotrexate. 
     
     
         22 . The method of  claim 8 , further comprising irradiating the target cells. 
     
     
         23 . The method of  claim 15 , further comprising contacting the target cells with a therapeutic agent. 
     
     
         24 . The method of  claim 23 , wherein the therapeutic agent is a chemotherapeutic agent. 
     
     
         25 . The method of  claim 23 , wherein the therapeutic agent is selected from the group consisting of bleomycin, carboplatin, chlorambucil, cisplatin, colchicine, cyclophosphamide, daunorubicin, actinomycin, diethylstilbestrol doxorubicin, etoposide, 5-fluorouracil, floxuridine, melphalan, methotrexate, mitomycin, 6-mercaptopurine, teniposide, 6-thioguanine, vincristine and vinblastine. 
     
     
         26 . The method of  claim 23 , wherein the therapeutic agent is doxorubicin. 
     
     
         27 . The method of  claim 23 , wherein the therapeutic agent is paclitaxel. 
     
     
         28 . The method of  claim 23 , wherein the therapeutic agent is methotrexate. 
     
     
         29 . The method of  claim 15 , further comprising irradiating the target cells. 
     
     
         30 . The method of  claim 16 , further comprising irradiating the target cells. 
     
     
         31 . A composition comprising a therapeutic amount of the antibody of  claim 1  and a pharmaceutically acceptable carrier. 
     
     
         32 . A method of treating a subject with cancer, comprising administering to the subject a therapeutic amount of the antibody of  claim 1 , wherein the therapeutic amount of the antibody selectively induces apoptosis of cancer cells in the subject. 
     
     
         33 . The method of  claim 32 , further comprising administering to the subject a therapeutic quantity of an antibody which specifically binds a TRAIL receptor DR5, wherein said second antibody, in its soluble form, has in vivo and in vitro apoptosis-inducing activity in malignant cells expressing DR5. 
     
     
         34 . The method of  claim 32 , further comprising administering to the subject a therapeutic agent. 
     
     
         35 . The method of  claim 34 , wherein the therapeutic agent is a chemotherapeutic agent. 
     
     
         36 . The method of  claim 34 , wherein the therapeutic agent is selected from the group consisting of bleomycin, carboplatin, chlorambucil, cisplatin, colchicine, cyclophosphamide, daunorubicin, actinomycin, diethylstilbestrol doxorubicin, etoposide, 5-fluorouracil, floxuridine, melphalan, methotrexate, mitomycin, 6-mercaptopurine, teniposide, 6-thioguanine, vincristine and vinblastine. 
     
     
         37 . The method of  claim 34 , wherein the therapeutic agent is doxorubicin. 
     
     
         38 . The method of  claim 34 , wherein the therapeutic agent is methotrexate. 
     
     
         39 . The method of  claim 32 , further comprising administering to the subject irradiation treatment. 
     
     
         40 . The method of  claim 33 , further comprising administering to the subject a therapeutic agent. 
     
     
         41 . The method of  claim 40 , wherein the therapeutic agent is a chemotherapeutic agent. 
     
     
         42 . The method of  claim 40 , wherein the therapeutic agent is selected from the group consisting of bleomycin, carboplatin, chlorambucil, cisplatin, colchicine, cyclophosphamide, daunorubicin, actinomycin, diethylstilbestrol doxorubicin, etoposide, 5-fluorouracil, floxuridine, melphalan, methotrexate, mitomycin, 6-mercaptopurine, teniposide, 6-thioguanine, vincristine and vinblastine. 
     
     
         43 . The method of  claim 40 , wherein the therapeutic agent is doxorubicin. 
     
     
         44 . The method of  claim 40 , wherein the therapeutic agent is paclitaxel. 
     
     
         45 . The method of  claim 40 , wherein the therapeutic agent is methotrexate. 
     
     
         46 . The method of  claim 33 , further comprising administering to the subject irradiation treatment. 
     
     
         47 . The method of  claim 34 , further comprising administering to the subject irradiation therapy. 
     
     
         48 . A method of treating a subject having an inflammatory or autoimmune disease, comprising administering to the subject a therapeutic amount of the antibody of claim  1 , wherein the therapeutic amount of the antibody selectively induces apoptosis of target cells having DR4 receptors. 
     
     
         49 . The method of  claim 48 , wherein the target cells are activated immune cells. 
     
     
         50 . The method of  claim 49 , wherein the activated immune cells are activated lymphocytes. 
     
     
         51 . The method of  claim 48 , wherein the target cells are rheumatoid arthritis synovial cells. 
     
     
         52 . The method of  claim 48 , further comprising administering to the subject a therapeutic amount of a second antibody that induces apoptosis of the target cells. 
     
     
         53 . The method of  claim 48 , further comprising administering to the subject a therapeutic amount of a therapeutic agent used in the treatment of inflammatory disease or autoimmune disease. 
     
     
         54 . The method of  claim 48 , further comprising administering to the subject a therapeutic amount of a therapeutic agent used in the treatment of inflammatory disease or autoimmune disease.