Il-15 and il-15ralpha sushi domain based modulokines

Abstract

The present invention relates to an immunocytokine comprising (a) a conjugate, and (b) an immunomodulatory antibody or a fragment thereof directly or indirectly linked by covalence to said conjugate, wherein said conjugate comprises (i) a polypeptide comprising the amino acid sequence of the interleukin 15 or derivatives thereof, and a polypeptide comprising the amino acid sequence of the sushi domain of the IL-15Rα or derivatives thereof; and uses thereof.

We claim: 
     
         1 . An immunocytokine comprising a conjugate and an immunomodulatory antibody or antigen binding fragment thereof,
 wherein said conjugate comprises
 (i) an interleukin 15-containing polypeptide comprising an amino acid sequence having at least 92.5% identity to SEQ ID NO:3, and 
 (ii) an IL-15Rα sushi domain-containing polypeptide comprising an amino acid sequence having at least 92% identity to SEQ ID NO:8; and 
   wherein said immunomodulatory antibody or antigen binding fragment thereof is capable of binding PD-1, PD-L1, or PD-L2, and wherein said antibody or antigen binding fragment thereof is a PD-1, PD-L1, or PD-L2 antagonist, and   wherein the conjugate and immunomodulatory antibody or antigen binding fragment thereof are covalently linked.   
     
     
         2 . The immunocytokine of  claim 1 , wherein the interleukin 15 polypeptide amino acid sequence has at least 96% identity, or 99% identity to SEQ ID NO:3. 
     
     
         3 . The immunocytokine of  claim 1 , wherein the interleukin 15 polypeptide amino acid sequence has at least one amino acid substitution selected from L45D, L45E, S51D, L52D, N72D, N72E, N72A, N72S, N72Y and N72P of SEQ ID NO: 3. 
     
     
         4 . The immunocytokine of  claim 1 , wherein conjugate is an IL-15 agonist or superagonist. 
     
     
         5 . The immunocytokine of  claim 1 , wherein the IL-15Rα sushi domain-containing polypeptide has at least 96% identity, or at least 98% identity to SEQ ID NO:8. 
     
     
         6 . The immunocytokine of  claim 5 , wherein the IL-15Rα sushi domain-containing polypeptide comprises SEQ ID NO:9. 
     
     
         7 . The immunocytokine of  claim 1 , wherein the IL-15Rα sushi domain-containing polypeptide further comprises the hinge domain of IL-15Rα, and the polypeptide comprises an amino acid sequence having at least 93% identity to SEQ ID NO:12. 
     
     
         8 . The immunocytokine of  claim 1 , wherein (i) and (ii) in the conjugate are either non-covalently linked or covalently linked 
     
     
         9 . The immunocytokine of  claim 8 , wherein the wherein (i) and (ii) in the conjugate are covalently linked using a bifunctional protein coupling agent selected from the group consisting of N-succinimidyl (2-pyridyldithio) propionate (SPDP), succinimidyl (N-maleimidomethyl) cyclohexane-1-carboxylate, iminothiolane (IT), bifunctional imidoesters, esters, aldehydes, bis-azido compounds, bis-diazonium compounds, diisocyanates, and bis-active fluorine compounds. 
     
     
         10 . The immunocytokine of  claim 1 , wherein (i) and (ii) in the conjugate are covalently linked as a fusion protein. 
     
     
         11 . The immunocytokine of  claim 10 , wherein the conjugate that is a fusion protein comprising (iii) a flexible linker covalently linking (i) and (ii), the flexible linker optionally consisting of 15-25 amino acids. 
     
     
         12 . The immunocytokine of  claim 11 , wherein the flexible linker has an amino acid sequence selected from the group consisting of SEQ ID NO: 13, SEQ ID NO: 14, and SEQ ID NO: 15. 
     
     
         13 . The immunocytokine of  claim 10 , wherein the conjugate that is a fusion protein has an amino acid sequence according to SEQ ID NO:16 or SEQ ID NO:17. 
     
     
         14 . The immunocytokine of  claim 1 , wherein said antibody is selected in the group consisting of nivolumab (BMS-936558 or MDX1106), Merck 3745 (MK-3475 or SCH-900475), CT-01 1 (hBAT or hBAT-1), lambrolizumab, AMP514, MDX-1 105, and YW243.55.S70 (BMS-936559) or the antibody fragment is selected from the group consisting of fragments of said antibodies. 
     
     
         15 . The immunocytokine of  claim 1 , wherein said conjugate (i) is in a C-terminal position relative to (ii). 
     
     
         16 . The immunocytokine of  claim 1 , wherein said conjugate is in a C-terminal position or an N-terminal position relative to the antibody or fragment thereof. 
     
     
         17 . The immunocytokine of  claim 16 , wherein said conjugate is in a C-terminal position relative to a heavy chain constant region of the antibody or fragment thereof. 
     
     
         18 . The immunocytokine of  claim 16 , wherein said conjugate is linked to the antibody or fragment thereof by a linker amino acid sequence. 
     
     
         19 . The immunocytokine of  claim 16 , wherein the immunocytokine is a fusion protein and wherein the conjugate and the antibody or fragment thereof are not separated by any linker. 
     
     
         20 . A nucleic acid encoding the immunocytokine of  claim 1 . 
     
     
         21 . A vector comprising the nucleic acid of  claim 19 . 
     
     
         22 . A host cell comprising the vector of  claim 20 , optionally being an animal cell. 
     
     
         23 . A pharmaceutical composition comprising the immunocytokine of  claim 1  or a nucleic acid or vector encoding the immunocytokine of  claim 1 , optionally comprising a pharmaceutically acceptable carrier, optionally configured for treating cancer in a subject, wherein optionally the cancer is advanced cancer, and optionally the advanced cancer is Renal Cancer Carcinoma (RCC). 
     
     
         24 . A method for treating cancer or inhibiting tumor growth in a subject comprising administering to a subject the immunocytokine of  claim 1 .