JPWO2019191295A5 - - Google Patents

Claims (29)

(A) First polypeptide containing the interleukin-2 (IL2) polypeptide; and (b) Second polypeptide containing the extracellular domain of the interleukin-2 receptor alpha (IL2Rα) polypeptide;
Including, here,
(I) The extracellular domain of the IL2Rα polypeptide has at least one less glycosylation compared to the extracellular domain of native IL2Rα (SEQ ID NO: 7); and / or (ii) the IL2 polypeptide is native IL2 (i). At least one less glycosylation compared to SEQ ID NO: 2);
A fusion protein that has IL2 activity. The fusion protein of claim 1, wherein the IL2 polypeptide has at least one less glycosylation as compared to native IL2 (SEQ ID NO: 2). The first polypeptide is at least about 60%, at least about 70%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97% relative to SEQ ID NO: 2. The fusion protein of claim 1 or 2, comprising an amino acid sequence having at least about 98%, at least about 99% or about 100% sequence identity . The second polypeptide is at least about 60%, at least about 70%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97% with respect to SEQ ID NO: 12. , The fusion protein of any of claims 1-3 , comprising an amino acid sequence having at least about 98%, at least about 99% or about 100% sequence identity . The fusion protein according to any one of claims 1 to 4 , wherein the extracellular domain of the IL2Rα polypeptide having at least one less glycosylation comprises a mutation that eliminates glycosylation as compared to the native IL2Rα polypeptide . Amino acids 192 to 219, amino acids 167 to 219, amino acids 168 to 219, amino acids 169 to 219, amino acids 170 to 219, amino acids 171 to 219, amino acids 172 to 219, amino acids 173 to 219, corresponding to SEQ ID NO: 7. Amino acids 174 to 219, amino acids 175 to 219, amino acids 176 to 219, amino acids 177 to 219, amino acids 178 to 219, amino acids 179 to 219, amino acids 180 to 219, amino acids 181 to 219, amino acids 182 to 219, amino acids 183 to 219, 5. A deletion of amino acids 184-219, amino acids 185-219, amino acids 186-219, amino acids 187-219, amino acids 188-219, amino acids 189-219, amino acids 190-219, or amino acids 191-219. The fusion protein described in . The fusion protein of claim 6 , wherein the second polypeptide comprises the amino acid sequence set forth in SEQ ID NO: 11 or SEQ ID NO: 12 . (A) The mutation is one or more substitutions, where the one or more substitutions are amino acid N49, amino acid N68, amino acid T74, amino acid T85, amino acid T197, amino acid T203, amino acid T208 and amino acid T216 or any combination thereof. Yes, where the amino acid position corresponds to SEQ ID NO: 7 ; or
(B) The mutation is one or more substitutions, where the one or more substitutions are amino acid S50, amino acid S51, amino acid T69, amino acid T70, amino acid C192 or any combination thereof, where the amino acid position is Corresponding to SEQ ID NO: 7,
The fusion protein according to claim 5 . The fusion protein comprises a mutation within the first polypeptide, wherein the mutation within the first polypeptide is one or more substitutions at amino acids T3 and / or C125 as compared to SEQ ID NO: 2. Item 2. The fusion protein according to Item 2. The fusion of claim 2 , wherein the fusion protein comprises a mutation within the first polypeptide, wherein the mutation within the first polypeptide is a deletion in amino acid A1 as compared to SEQ ID NO: 2. protein. The fusion protein according to any one of claims 1 to 10 , wherein the fusion protein is enzymatically or chemically deglycosylated. The fusion protein according to any one of claims 1 to 11 , further comprising a linker fused in frame between the first polypeptide and the second polypeptide. The fusion protein of claim 12 , wherein the linker is a glycine / serine linker. 13. The fusion protein of claim 13 , wherein the glycine / serine linker comprises the amino acid sequence of (GGGS) ₃ . The fusion protein according to any one of claims 1 to 14 , wherein the fusion protein further comprises a heterologous moiety fused to the first polypeptide and / or the second polypeptide. The fusion protein according to claim 15 , wherein the heterologous moiety is an extended half-life moiety. Heterogeneous parts are albumin, immunoglobulin constant region or part thereof, immunoglobulin-binding polypeptide, immunoglobulin G (IgG), albumin-binding polypeptide (ABP), PASylation part, HESylation part, XTEN, pegged part, Fc region , The fusion protein according to claim 16 , which comprises any combination thereof. The fusion protein according to claim 1, wherein the fusion protein comprises the amino acid sequence set forth in SEQ ID NO: 13 to SEQ ID NO: 70, SEQ ID NO: 202, or SEQ ID NO: 203. The fusion protein according to claim 1, wherein the fusion protein comprises the amino acid sequence set forth in SEQ ID NO: 16. The fusion protein according to any one of claims 1 to 19 , which is a monomer or a dimer . A nucleic acid encoding the fusion protein according to any one of claims 1 to 20 . A vector comprising the nucleic acid of claim 21 . A host cell comprising the nucleic acid of claim 21 . (A) the fusion protein according to any one of claims 1 to 20 , the nucleic acid according to claim 21 , the vector according to claim 22 or the host cell according to claim 23 ; and (b) pharmaceutically. A pharmaceutical composition comprising an acceptable additive. The fusion protein according to any one of claims 1 to 20 , the nucleic acid according to claim 21 , the vector according to claim 22 , the host cell according to claim 23 , or the pharmaceutical composition according to claim 24. , And instructions for administering the fusion protein to subjects in need thereof. A method for producing a fusion protein, comprising culturing the host cell according to claim 23 under appropriate conditions and recovering the fusion protein. The fusion protein of any of claims 1-20 , the nucleic acid of claim 21 , the vector of claim 22 , for use in the treatment of a disease or disorder in a subject in need of treatment. The host cell according to claim 23 , or the pharmaceutical composition according to claim 24 . Claim that the disease or disorder is an inflammatory disease, autoimmune disease , or infectious disease, wherein the infectious disease is due to a pathogenic virus, pathogenic bacterium, pathogenic fungus, or pathogenic parasite. 27. The fusion protein, nucleic acid, vector, host cell, or pharmaceutical composition according to 27 . Inflammatory or autoimmune diseases are type I diabetes, multiple sclerosis, rheumatoid arthritis, celiac disease, systemic erythematosus, lupus nephritis, cutaneous lupus, juvenile idiopathic arthritis, Crohn's disease, ulcerative colitis or systemic Sclerosis, transplant-to-host disease, psoriasis, alopecia, HCV-induced vasculitis, Sjogren's syndrome, scoliosis, tonic spondylitis, Bechet's disease, Wegener's granulomas, hyperan disease, autoimmune hepatitis, sclerosing bile duct 28. The fusion protein, nucleic acid, vector, host cell, or pharmaceutical composition of claim 28 , selected from the group consisting of flame, Gugel-Sjogren and macrophage activation syndrome.