Biologic compounds

Abstract

The present invention relates to amino acid sequences that are directed against TRAIL cell surface receptor 2 (herein also “DR5”), as well as to compounds or constructs thereof, and in particular proteins and polypeptides and nucleotides that encode them (referred to herein in their entirety as “NB agents”) and fragments thereof, and pharmaceutically effective variants thereof, and their use in the diagnosis and treatment of DR5 associated diseases and disorders.

What is claimed is: 
     
         1 . A method of inducing apoptosis in a cell or tissue in a patient, comprising the method of administering to the patient an isolated polypeptide comprising at least one monomer of a single variable domain comprising the sequence of the complementarity determining region 3 (CDR3) sequence of SEQ ID NO: 66, the sequence of the complementarity determining region 2 (CDR2) sequence of SEQ ID NO: 53, and the sequence of the complementarity determining region 1 (CDR1) sequence of SEQ ID NO: 43, Wherein the polypeptide is capable of binding to DR5 on a surface of the cell or tissue and inducing apoptosis. 
     
     
         2 . The method of  claim 1 , wherein the cell or tissue is a cancer or proliferative disease. 
     
     
         3 . The method of  claim 2 , wherein the cell or tissue is selected from:
 a) one or more solid cancers selected from primary and metastatic cancers, renal cell carcinoma, and cancers of the lung, pancreas, hematopoietic malignancy, glioma, astrocytoma, mesothelioma, colorectal cancers, prostate cancer, osteosarcoma, melanoma, lymphoma lymphoma, breast cancer, endometrial cancer, liver cancer, gastric cancer, skin cancer, ovarian cancer and squamous cell cancers of any origin, squamous cell cancers of the lung, head and neck, breast, thyroid, cervix, skin, and/or esophageal; and   b) one or more liquid cancers selected from leukemias, a T-cell leukemia such as acute T-cell leukemia (T-ALL), acute B-cell leukemia (B-ALL), chronic myelogenous leukemia (CML), acute myelogenous leukemia (AML), plasma cell myeloma and multiple myeloma (MM); and   c) one or more non-cancer indications, said indications comprising one or more of inflammatory and autoimmune diseases, systemic lupus erythematosus, Hashimoto's disease, rheumatoid arthritis, graft-versus-host disease, Sjogren's syndrome, pernicious anemia, Addison disease, scleroderma, Goodpasture's syndrome, Crohn's disease, autoimmune hemolytic anemia, sterility, myasthenia gravis, multiple sclerosis, Basedow's disease, thrombotic throbocytopenia, thrombopenia purpurea, insulin-dependent diabetes mellitus, allergy; asthma, atopic disease; arteriosclerosis; myocarditis; cardiomyopathy; globerula nephritis; and hypoplastic anemia.   
     
     
         4 . The method of  claim 2 , wherein the proliferative disease is pancreatic cancer. 
     
     
         5 . The method of  claim 2 , wherein the proliferative disease is T-ALL. 
     
     
         6 . The method of  claim 2 , wherein the proliferative disease is mesothelioma. 
     
     
         7 . The method of  claim 2 , wherein the proliferative disease is squamous cell carcinoma of any tissue origin. 
     
     
         8 . The method of  claim 2 , wherein the proliferative disease is AML. 
     
     
         9 . The method of  claim 2 , wherein the proliferative disease is melanoma. 
     
     
         10 . The method of  claim 2 , wherein the proliferative disease is myeloma. 
     
     
         11 . The method of  claim 1 , wherein the single variable domain comprises:
 (a) the sequence of SEQ ID NO: 30; or   (b) the sequence of SEQ ID NO: 30 wherein one or more of the amino acid positions 1-30, 36-49, 66-97 and/or 112-122 have been substituted, modified or deleted and/or wherein one or more amino acids have been inserted between any amino acids at 1-30, 36-49, 66-97 and/or 112-122.   
     
     
         12 . The method of  claim 1 , wherein the isolated polypeptide comprises at least three, at least four, or at least five monomeric subunits of a single variable domain that specifically binds to human DR5, wherein said subunits are linked by a linker, and wherein the variable domain comprises the sequence of the complementarity determining region 3 (CDR3) sequence of SEQ ID NO: 66, the sequence of the complementarity determining region 2 (CDR2) sequence of SEQ ID NO: 53, and the sequence of the complementarity determining region 1 (CDR1) sequence of SEQ ID NO: 43. 
     
     
         13 . The method of  claim 12 , wherein said single variable domain comprises the sequence of SEQ ID NO: 30. 
     
     
         14 . The method of  claim 13 , wherein the single variable domain comprises the amino acid sequence of SEQ ID NO: 30 wherein one or more of the amino acid positions 1-30, 36-49, 66-97 and/or 112-122 have been substituted, modified or deleted, and/or wherein one or more amino acids have been inserted between any amino acids at 1-30, 36-49, 66-97 and/or 112-122. 
     
     
         15 . The method of  claim 1 , wherein the polypeptide comprises the amino acid sequence of SEQ ID NO: 5, 20, 21, 22, 30, 31, 32, 33, or 87. 
     
     
         16 . The method of  claim 1 , wherein the polypeptide has:
 (a) the amino acid sequence of SEQ ID NO: 30; or   (b) the amino acid sequence of SEQ ID NO: 30, in which one or more of the amino acid positions 11, 37, 44, 45, 47, 83, 84, 112, 113 and 117 of SEQ ID NO: 30 are humaneered, substituted, modified or deleted and/or wherein one or more amino acids have been inserted between any amino acids at 1-30, 36-49, 66-97 and/or 112-122, wherein the isolated polypeptide specifically binds to human DR5.   
     
     
         17 . The method of  claim 1 , wherein the polypeptide has:
 (a) the sequence of SEQ ID NO: 30; and/or   (b) the sequence of SEQ ID NO: 30 wherein one or more of the amino acid positions 1-30, 36-49, 66-97 and/or 112-122 have been substituted, modified or deleted, and/or wherein one or more amino acids have been inserted between any amino acids at 1-30, 36-49, 66-97 and/or 112-122, and wherein the isolated polypeptide is capable of enhancing cell apoptosis and wherein the isolated polypeptide specifically binds to human DR5.   
     
     
         18 . The method according to  claim 17 , wherein the polypeptide comprises at least three, four, five or more copies of the sequence of SEQ ID NO: 30 wherein one or more of the amino acid positions 1-30, 36-49, 66-97 and/or 112-122 have been substituted, modified or deleted, and/or wherein one or more amino acids have been inserted between any amino acids at 1-30, 36-49, 66-97 and/or 112-122, and wherein the copies constitute monovalent binding polypeptides that are all directed against the same binding region of human DR5. 
     
     
         19 . The method of  claim 1 , wherein the polypeptide comprises at least three, four, five or more copies of the sequence of SEQ ID NO: 30 wherein one or more of the amino acid positions 1-30, 36-49, 66-97 and/or 112-122 have been substituted, modified or deleted and/or wherein one or more amino acids have been inserted between any amino acids at 1-30, 36-49, 66-97 and/or 112-122, wherein the isolated polypeptide is capable of enhancing cell apoptosis and wherein the copies constitute monovalent binding polypeptides that are all directed against the same binding region of human DR5, and wherein the said isolated polypeptide specifically binds to human DR5. 
     
     
         20 . The method of  claim 1 , wherein the polypeptide comprises the CDR1 sequence of SEQ ID NO: 43, the CDR2 sequence of SEQ ID NO: 53, and the CDR3 sequence of SEQ ID NO: 66, wherein the polypeptide is capable of contacting, within 5 Angstroms, amino acids H86, S88, E89, D90, D93, I95, K98, Q101, D102, L111, F112, R115, and R118 of a DR5 polypeptide of SEQ ID NO: 89. 
     
     
         21 . The method of  claim 1 , wherein the polypeptide comprises the amino acid sequence of SEQ ID NO: 30.