FUSION PROTEINS COMPRISING A BINDING PROTEIN AND AN INTERLEUKIN-15 POLYPEPTIDE HAVING A REDUCED AFFINITY FOR IL15Ra AND THERAPEUTIC USES THEREOF

Abstract

The present invention relates to fusion proteins comprising a binding protein and an IL-15 polypeptide as well as uses thereof, pharmaceutical compositions comprising such fusion proteins and a method for producing such fusion proteins.

1 . A method of treating a disease comprising administering a therapeutically effective amount of the fusion protein to a subject,
 the fusion protein comprising:   a) a binding protein comprising at least one binding site, wherein the binding site binds to an antigen associated with a target cell; and   b) an IL-15 polypeptide, wherein the IL-15 polypeptide comprises at least one amino acid substitution at one or more positions corresponding to position(s) 92, 94, 95, 97, 98, 114, and/or 115 of the amino acid sequence shown in SEQ ID NO:1;   thereby having a reduced affinity for IL-15Rα compared to the affinity of wild-type IL-15 of SEQ ID NO: 1 (Uniprot number: P40933-1).   
     
     
         2 . The method protein of  claim 1 , wherein the binding protein is selected from the group consisting of an antibody, a divalent antibody fragment, a monovalent antibody fragment, or a proteinaceous binding molecule with antibody-like binding properties. 
     
     
         3 . The method of  claim 1 , wherein the target cell expresses a tumor associated antigen (TAA) and/or an antigen associated with autoimmune diseases. 
     
     
         4 . The method of  claim 3 , wherein the TAA is selected from the group consisting of CD19, CD20, CD10, CD21, CD22, CD25, CD30, CD33, CD34, CD37, CD38, CD44v6, CD45, CDw52, Fms-like tyrosine kinase 3 (FLT-3, CD135), c-Kit (CD117), CSF1R, (CD115), CD123, CD133, PDGFR-α (CD140a), PDGFR-β (CD140b), chondroitin sulfate proteoglycan 4 (CSPG4, melanoma-associated chondroitin sulfate proteoglycan), Muc-1, EGFR, de2-7-EGFR, EGFRvIII, Folate blocking protein, Her2neu, Her3, PSMA, PSCA, PSA, TAG-72, HLA-DR, IGFR, CD133, IL3R, fibroblast activating protein (FAP), Carboanhydrase IX (MN/CA IX), Carcinoembryonic antigen (CEA), EpCAM, CDCP1, Derlin1, Tenascin, frizzled 1-10, the vascular antigens VEGFR2 (KDR/FLK1), VEGFR3 (FLT4, CD309), Endoglin, CLEC14, Tem1-8, Tie2, mesothelin, epithelial glycoprotein 2 (EGP2), epithelial glycoprotein 40 (EGP40), cancer antigen 72-4 (CA72-4), interleukin 13 receptor alpha-2 subunit, IL13Rα2, Ig kappa light chain (κ), GD3-ganglioside (GD3), GD2-ganglioside (GD2), acetylated variants of GD2 and GD3, CD171, NCAM, alpha folate receptor (αFR), Lewis (Y), fetal acetylcholine receptor (FAR), avian erythroblastic leukemia viral oncogene homolog 3 (ERBB3), avian erythroblastic leukemia viral oncogene homolog 4 (ERBB4), avian erythroblastic leukemia viral oncogene homolog 2 (ERBB2), hepatocyte growth factor receptor (HGFR/c-Met), claudin 18.2, claudin 3, claudin 4, claudin 1, claudin 12, claudin 2, claudin 5, claudin 8, claudin 7, claudin 6, membrane bound CEA, Robo4, CD138, tenascin and the extra domain-B of fibronectin. 
     
     
         5 . The method of  claim 3 , wherein the target cell expresses an antigen associated with autoimmune diseases, which antigen is selected from the group consisting of CD20, CD22, CD52 and TNFR, CD19, CD25, CD40. 
     
     
         6 . The method of  claim 1 , wherein the target cell is a tumor/cancer cell and/or a B cell. 
     
     
         7 . The method of  claim 1 , wherein the at least one amino acid substitution is selected from the group consisting of L92D, E94K, L95D, V97D, I98D, L114D, L114E, I115D, I115E. 
     
     
         8 . The method of  claim 1 ,
 (i) wherein the IL-15 polypeptide does not bind to IL-15Rα; and/or   (ii) wherein the IL-15 polypeptide binds to IL-2/IL-15Rβγ.   
     
     
         9 . The method of  claim 1 , wherein the IL-15 polypeptide comprises at least an amino acid sequence as shown in SEQ ID NO: 4, which comprises at least one amino acid substitution at one or more positions. at least one amino acid substitution at one or more positions corresponding to position(s) 44, 46, 47, 49, 50, 66, and/or 67 of the amino acid sequence shown in SEQ ID NO: 4. 
     
     
         10 . The method of  claim 1 , wherein the fusion protein further comprises a linker. 
     
     
         11 . The method of  claim 1 , wherein the disease is a proliferatory or autoimmune disease. 
     
     
         12 . The method of  claim 11 , wherein the disease is a proliferatory disease selected from the group consisting of adrenal cancer, anal cancer, bile duct cancer, bladder cancer, bone cancer, brain and spinal cord tumors, breast cancer, Castleman disease, cervical cancer, colon cancer, endometrial cancer, esophagus cancer, Ewing family of tumors, eye cancer, gallbladder cancer, gastrointestinal carcinoid tumors, gastrointestinal stromal tumor (GIST), gestational trophoblastic disease, Hodgkin disease, Kaposi sarcoma, kidney cancer, laryngeal and hypopharyngeal cancer, leukemia, acute lymphocytic leukemia (ALL), acute myeloid leukemia (AML), chronic lymphocytic leukemia (CLL), chronic myeloid leukemia (CML), chronic myelomonocytic leukemia (CMML), liver cancer, lung cancer, non-small cell lung cancer, small cell lung cancer, lung carcinoid tumor, lymphoma, lymphoma of the skin, malignant mesothelioma, multiple myeloma, myelodysplastic syndrome, nasal cavity and paranasal sinus cancer, nasopharyngeal cancer, neuroblastoma, non-Hodgkin lymphoma, oral cavity and oropharyngeal cancer, osteosarcoma, ovarian cancer, pancreatic cancer, penile cancer, pituitary tumors, prostate cancer, rectum cancer, retinoblastoma, rhabdomyosarcoma, salivary gland cancer, sarcoma, skin cancer, basal and squamous cell cancer, melanoma, merkel cell cancer, small intestine cancer, stomach cancer, testicular cancer, thymus cancer, thyroid cancer, uterine sarcoma, vaginal cancer, vulvar cancer, Waldenstrom macroglobulinemia, or Wilms tumor. 
     
     
         13 . The method of  claim 11 , wherein the disease is an autoimmune disease selected from the group consisting of Systemic lupus erythematosus (SLE), Goodpasture's syndrome, Sarcoidosis, Scleroderma, Rheumatoid arthritis, Dermatomyositis, Sjögren's Syndrome, Scleroderma, Dermatomyositis, Psoriasis, Vitiligo, Alopecia areata, Type 1 diabetes mellitus, Autoimmune pancreatitis, Hashimoto's thyroiditis, Addison's disease, Multiple sclerosis, Myasthenia gravis, Polyarteritis nodosa, Idiopathic thrombocytopenic purpura, Hemolytic anemia, Antiphospholipid antibody syndrome, Pernicious anemia, Gastrointestinal diseases, Celiac disease, Inflammatory bowel disease, Autoimmune hepatitis or Primary biliary cirrhosis. 
     
     
         14 . The method of  claim 1 , wherein the subject is a vertebrate 
     
     
         15 . The method of  claim 14 , wherein the vertebrate is a human being. 
     
     
         16 . The method of  claim 1 , wherein side effects of the treatment are reduced. 
     
     
         17 . The method of  claim 16 , wherein the side effects include at least one of infusion reaction, elevated temperature/fever, dypnoe, circulatory system problems, immunogenicity, hypersensitivity reactions, immunosuppression, infections, anemia, autoimmune haemolytic anaemia, leukopenia, thrombopenia, pancytopenia, cytopenia, worsening heart failure, tumor lysis, cytokine release syndrome, thyroid disorders, cardiotoxicity, local skin reaction, elevated liver transaminases, hypotension, serum sickness, mucocutaneous reactions, hepatitis reactivation, progressive multifocal leukoencephalopathy (PML), renal toxicity, cardiac arrhythmias.