T cell recruiting polypeptides based on tcr alpha/beta reactivity

Abstract

T cell recruiting polypeptides are provided that bind the constant domain of TCR on a T cell. The polypeptides can be used in methods for treatment of cancers.

1 .- 81 . (canceled) 
     
     
         82 . A process for the production of the polypeptide, said process comprising
 culturing a host cell transformed or transfected with a nucleic acid or nucleic acid sequence encoding a polypeptide comprising a first and a second immunoglobulin single variable domain (ISV), wherein
 said first ISV specifically binds to the constant domain of the T cell receptor (TCR) present on a T cell; 
 said second ISV specifically binds to a first antigen on a target cell; 
 wherein said first antigen is different from said TCR; and 
 wherein said target cell is different from said T cell 
   under conditions allowing the expression of the polypeptide, and   recovering the produced polypeptide from the culture.   
     
     
         83 .- 86 . (canceled) 
     
     
         87 . A method for the treatment or amelioration of a disease selected from the group consisting of a proliferative disease, an inflammatory disease, an infectious disease and an autoimmune disease, comprising
 administering to a subject in need thereof a polypeptide comprising a first and a second immunoglobulin single variable domain (ISV), wherein
 said first ISV specifically binds to the constant domain of the T cell receptor (TCR) present on a T cell; 
 said second ISV specifically binds to a first antigen on a target cell; 
 wherein said first antigen is different from said TCR; and 
 wherein said target cell is different from said T cell. 
   
     
     
         88 . The method according to  claim 87 , wherein said proliferative disease is cancer. 
     
     
         89 . The method according to  claim 88 , wherein said cancer is chosen from the group consisting of carcinomas, gliomas, mesotheliomas, melanomas, lymphomas, leukemias, adenocarcinomas: breast cancer, ovarian cancer, cervical cancer, glioblastoma, multiple myeloma (including monoclonal gammopathy of undetermined significance, asymptomatic and symptomatic myeloma), prostate cancer, and Burkitt's lymphoma, head and neck cancer, colon cancer, colorectal cancer, non-small cell lung cancer, small cell lung cancer, cancer of the esophagus, stomach cancer, pancreatic cancer, hepatobiliary cancer, cancer of the gallbladder, cancer of the small intestine, rectal cancer, kidney cancer, bladder cancer, prostate cancer, penile cancer, urethral cancer, testicular cancer, vaginal cancer, uterine cancer, thyroid cancer, parathyroid cancer, adrenal cancer, pancreatic endocrine cancer, carcinoid cancer, bone cancer, skin cancer, retinoblastomas, Hodgkin's lymphoma, non-Hodgkin's lymphoma, Kaposi's sarcoma, multicentric Castleman's disease or AIDS-associated primary effusion lymphoma, neuroectodermal tumors, rhabdomyosarcoma; as well as any metastasis of any of the above cancers. 
     
     
         90 . The method according to  claim 88 , wherein the treatment is a combination treatment. 
     
     
         91 . (canceled) 
     
     
         92 . The method according to  claim 87 , wherein said first ISV binds to the constant domain of a T cell receptor α (TCR-α) (SEQ ID NO: 348 and/or SEQ ID NO: 484) and/or the constant domain of the T cell receptor β (TCR-β) (SEQ ID NO: 349 and/or SEQ ID NO: 485), or polymorphic variants or isoforms thereof. 
     
     
         93 . The method according to  claim 87 , wherein said first ISV essentially consists of 4 framework regions (FR1 to FR4, respectively) and 3 complementarity determining regions (CDR1 to CDR3, respectively), in which:
 (i) CDR1 is chosen from the group consisting of:
 (a) SEQ ID NOs: 119-133; or 
 (b) amino acid sequences that have 4, 3, 2, or 1 amino acid(s) difference with the amino acid sequence of SEQ ID NO: 123 or with any of SEQ ID NOs: 119-133; and/or 
   (ii) CDR2 is chosen from the group consisting of:
 (c) SEQ ID NOs: 134-163; or 
 (d) amino acid sequences that have 4, 3, 2, or 1 amino acid(s) difference with the amino acid sequence of SEQ ID NO: 153 or with any of SEQ ID NOs: 134-163; and/or 
   (iii) CDR3 is chosen from the group consisting of:
 (e) SEQ ID NOs: 164-174; or 
 (f) amino acid sequences that have 4, 3, 2, or 1 amino acid(s) difference with the amino acid sequence of SEQ ID NO: 170 or with any of SEQ ID NOs: 164-174. 
   
     
     
         94 . The method according to  claim 87 , further comprising a third ISV, which specifically binds to a second antigen on a target cell, wherein said second antigen is different from said first antigen. 
     
     
         95 . The method according to  claim 87 , wherein said first antigen on a target cell is a tumour antigen, preferably a tumour associated antigen (TAA). 
     
     
         96 . The method according to  claim 94 , wherein said second antigen on a target cell is a tumour antigen, preferably a tumour associated antigen (TAA). 
     
     
         97 . The method according to  claim 94 , wherein said first antigen and said second antigen are present on the same target cell. 
     
     
         98 . The method according to  claim 94 , wherein said first antigen and said second antigen are present on different target cells. 
     
     
         99 . The method according to  claim 87 , further comprising a serum protein binding moiety. 
     
     
         100 . The method according to  claim 99 , wherein said serum protein binding moiety is an ISV binding serum albumin. 
     
     
         101 . The method according to  claim 87 , wherein said first ISV and said second ISV are linked via a linker. 
     
     
         102 . The method according to  claim 101 , wherein said linker is chosen from the group consisting of linkers of 5GS, 7GS, 9GS, 10GS, 15GS, 18GS, 20GS, 25GS, 30GS and 35GS (SEQ ID NOs: 376 to 385). 
     
     
         103 . The method according to  claim 87 , wherein said ISV is a Nanobody, a V HH , a humanized VHH, or a camelized V H . 
     
     
         104 . The method according to  claim 95 , wherein said TAA is chosen from the group consisting of Melanoma-associated Chondroitin Sulfate Proteoglycan (MCSP), Epidermal Growth Factor Receptor (EGFR), Fibroblast Activation Protein (FAP), MART-1, carcinoembryonic antigen (CEA), gp100, MAGE-1, HER-2, LewisY antigens, CD123, CD44, CLL-1, CD96, CD47, CD32, CXCR4, Tim-3, CD25, TAG-72, Ep-CAM, PSMA, PSA, GD2, GD3, CD4, CD5, CD19, CD20, CD22, CD33, CD36, CD45, CD52, CD147, growth factor receptors including ErbB3 and ErbB4, Cytokine receptors including Interleukin-2 receptor gamma chain (CD132 antigen), Interleukin-10 receptor alpha chain (IL-10R-A), Interleukin-10 receptor beta chain (IL-10R-B), Interleukin-12 receptor beta-1 chain (IL-12R-beta1), Interleukin-12 receptor beta-2 chain (IL-12 receptor beta-2), Interleukin-13 receptor alpha-1 chain (IL-13R-alpha-1) (CD213a1 antigen), Interleukin-13 receptor alpha-2 chain (Interleukin-13 binding protein), Interleukin-17 receptor (IL-17 receptor), Interleukin-17B receptor (IL-17B receptor), Interleukin 21 receptor precursor (IL-21R), Interleukin-1 receptor type I (IL-1R-1) (CD121a), Interleukin-1 receptor type II (IL-1R-beta) (CDw121b), Interleukin-1 receptor antagonist protein (IL-1ra), Interleukin-2 receptor alpha chain (CD25 antigen), Interleukin-2 receptor beta chain (CD122 antigen), Interleukin-3 receptor alpha chain (IL-3R-alpha) (CD123 antigen), CD30, IL23R, IGF-1R, IL5R, IgE, CD248 (endosialin), CD44v6, gpA33, Ron, Trop2, PSCA, claudin 6, claudin 18.2, CLEC12A, CD38, ephA2, c-Met, CD56, MUC16, EGFRvIII, AGS-16, CD27L, Nectin-4, SLITRK6, mesothelin, folate receptor, tissue factor, axl, glypican-3, CA9, Cripto, CD138, CD37, MUC1, CD70, gastrin releasing peptide receptor, PAP, CEACAM5, CEACAM6, CXCR7, N-cadherin, FXYD2 gamma a, CD21, CD133, Na/K-ATPase, mIgM (membrane-bound IgM), mIgA (membrane-bound IgA), Mer, Tyro2, CD120, CD95, CA 195, DR5, DR6, DcR3 and CAIX, and related polymorphic variants and isoforms. 
     
     
         105 . The method according to  claim 96 , wherein said TAA is chosen from the group consisting of Melanoma-associated Chondroitin Sulfate Proteoglycan (MCSP), Epidermal Growth Factor Receptor (EGFR), Fibroblast Activation Protein (FAP), MART-1, carcinoembryonic antigen (CEA), gp100, MAGE-1, HER-2, LewisY antigens, CD123, CD44, CLL-1, CD96, CD47, CD32, CXCR4, Tim-3, CD25, TAG-72, Ep-CAM, PSMA, PSA, GD2, GD3, CD4, CD5, CD19, CD20, CD22, CD33, CD36, CD45, CD52, CD147, growth factor receptors including ErbB3 and ErbB4, Cytokine receptors including Interleukin-2 receptor gamma chain (CD132 antigen), Interleukin-10 receptor alpha chain (IL-10R-A), Interleukin-10 receptor beta chain (IL-10R-B), Interleukin-12 receptor beta-1 chain (IL-12R-beta1), Interleukin-12 receptor beta-2 chain (IL-12 receptor beta-2), Interleukin-13 receptor alpha-1 chain (IL-13R-alpha-1) (CD213a1 antigen), Interleukin-13 receptor alpha-2 chain (Interleukin-13 binding protein), Interleukin-17 receptor (IL-17 receptor), Interleukin-17B receptor (IL-17B receptor), Interleukin 21 receptor precursor (IL-21R), Interleukin-1 receptor type I (IL-1R-1) (CD121a), Interleukin-1 receptor type II (IL-1R-beta) (CDw121b), Interleukin-1 receptor antagonist protein (IL-1ra), Interleukin-2 receptor alpha chain (CD25 antigen), Interleukin-2 receptor beta chain (CD122 antigen), Interleukin-3 receptor alpha chain (IL-3R-alpha) (CD123 antigen), CD30, IL23R, IGF-1R, IL5R, IgE, CD248 (endosialin), CD44v6, gpA33, Ron, Trop2, PSCA, claudin 6, claudin 18.2, CLEC12A, CD38, ephA2, c-Met, CD56, MUC16, EGFRvIII, AGS-16, CD27L, Nectin-4, SLITRK6, mesothelin, folate receptor, tissue factor, axl, glypican-3, CA9, Cripto, CD138, CD37, MUC1, CD70, gastrin releasing peptide receptor, PAP, CEACAM5, CEACAM6, CXCR7, N-cadherin, FXYD2 gamma a, CD21, CD133, Na/K-ATPase, mIgM (membrane-bound IgM), mIgA (membrane-bound IgA), Mer, Tyro2, CD120, CD95, CA 195, DR5, DR6, DcR3 and CAIX, and related polymorphic variants and isoforms. 
     
     
         106 . The method according to  claim 94 , wherein said first antigen and said second antigen are chosen from the group consisting of:
 EGFR as a first antigen and CEA as a second antigen;   CD19 as a first antigen and CD20 as a second antigen;   CD19 as a first antigen and CD22 as a second antigen;   CD123 as a first antigen and Tim-3 as a second antigen; and   CD123 as a first antigen and CD69 as a second antigen.