Modified soluble fgf receptor fc fusions method

Abstract

The invention relates to modified soluble FGF receptor Fc fusions comprising a fusion of a soluble fragment or domain of the FGF receptor part (targeting or binding moiety) with an Fc region of an immunoglobulin part (effector function moiety), having improved biological activity including ADCC/CDC activities, compositions containing them, and method of producing such modified soluble FGF receptor Fc fusion molecules.

1 . A modified soluble FGF receptor Fc fusion comprising a fusion of a soluble fragment or domain of a FGF receptor with an Fc region of an immunoglobulin, wherein at least the 5th N-glycosylation site of the FGF receptor moiety is occupied, and at most 45% of the N-glycans of said FGF receptor moiety have no sialyl group. 
     
     
         2 . The modified soluble FGF receptor Fc fusion of  claim 1 , wherein, in addition, the 3rd, 4th, 6th and 7th N-glycosylation sites of the FGF receptor moiety are occupied. 
     
     
         3 . The modified soluble FGF receptor Fc fusion of  claim 2 , wherein at least 7 N-glycosylation sites of the FGF receptor moiety are occupied. 
     
     
         4 . The modified soluble FGF receptor Fc fusion of  claim 3 , wherein all N-glycosylation sites of the FGF receptor moiety are occupied. 
     
     
         5 . The modified soluble FGF receptor Fc fusion of  claim 1 , wherein the average number of sialic acid per N-glycan of the FGF receptor moiety is 0.9 or above. 
     
     
         6 . The modified soluble FGF receptor Fc fusion of  claim 5 , wherein the average number of sialic acid per N-glycan of the FGF receptor moiety is 1.2 or above. 
     
     
         7 . (canceled) 
     
     
         8 . (canceled) 
     
     
         9 . The modified soluble FGF receptor Fc fusion of  claim 1 , wherein said fusion possesses ADCC and/or CDC activities. 
     
     
         10 . The modified soluble FGF receptor Fc fusion of  claim 1 , wherein the N-glycans of said fusion are 60-100% fucosylated. 
     
     
         11 . The modified soluble FGF receptor Fc fusion of  claim 1 , wherein said modified soluble FGF receptor Fc fusion comprises 3 mannose residues, a mean of 1.5 to 3.0 galactose residues, a mean of 3.5 to 5 of N-acetylglucosamine residues, and a mean of 0.6 to 1 fucose residues per molecule of glycan. 
     
     
         12 . The modified soluble FGF receptor Fc fusion of  claim 1 , wherein the N-glycans of said fusion are 0-60% fucosylated. 
     
     
         13 . The modified soluble FGF receptor Fc fusion of  claim 1 , wherein the FGF receptor is FGF receptor 1 (FGFR1) or FGF receptor 2 (FGFR2). 
     
     
         14 . The modified soluble FGF receptor Fc fusion of  claim 13 , wherein the FGF receptor is FGF receptor 1 isotype IIIc or FGF receptor 2 isotype IIIc. 
     
     
         15 . The modified soluble FGF receptor Fc fusion of  claim 1 , wherein the FGF receptor soluble domain has a sequence as set forth in SEQ ID NO: 4, or a sequence having an identity of at least 95% with the SEQ ID NO: 4. 
     
     
         16 . The modified soluble FGF receptor Fc fusion of  claim 1 , wherein the Fc portion has a sequence as set forth in SEQ ID NO: 6, or a sequence having an identity of at least 95% with the SEQ ID NO: 6. 
     
     
         17 . The modified soluble FGF receptor Fc fusion of  claim 1 , wherein said modified soluble FGF receptor Fc fusion further comprises a linker sequence of at least 3 amino acid residues. 
     
     
         18 . The modified soluble FGF receptor Fc fusion of  claim 17 , wherein the linker sequence is SAL (Ser-Ala-Leu). 
     
     
         19 . (canceled) 
     
     
         20 . The modified soluble FGF receptor Fc fusion of  claim 1 , wherein modified soluble FGF receptor Fc fusion further comprises the signal peptide of SEQ ID NO: 8. 
     
     
         21 . (canceled) 
     
     
         22 . (canceled) 
     
     
         23 . (canceled) 
     
     
         24 . (canceled) 
     
     
         25 . (canceled) 
     
     
         26 . A pharmaceutical composition comprising a modified fusion FGF receptor of  claim 1 . 
     
     
         27 . (canceled) 
     
     
         28 . (canceled) 
     
     
         29 . (canceled) 
     
     
         30 . (canceled) 
     
     
         31 . (canceled) 
     
     
         32 . A method for treating cancer comprising administering to a subject with said cancer a therapeutically effective amount of the modified soluble FGF receptor Fc fusion of  claim 1 . 
     
     
         33 . The method of  claim 32 , wherein the method further comprises a further therapeutic agent. 
     
     
         34 . The method of  claim 33 , wherein said further therapeutic agent is an anti-angiogenic agent or a chemotherapeutic agent. 
     
     
         35 . The method of  claim 34 , wherein said anti-angiogenic agent is a tumor necrosis factor, or an antagonist of an acidic or basic fibroblast growth factor (FGF), hepatocyte growth factor (HGF), tissue factor (TF), protein C, protein S, platelet-derived growth factor (PDGF), or HER2 receptor. 
     
     
         36 . The method of  claim 34 , wherein said chemotherapeutic agent is selected from the group consisting of: anti-microtubule agents; platinum coordination complexes; alkylating agents; antibiotic agents; topoisomerase II inhibitors; antimetabolites; topoisomerase I inhibitors; hormones and hormone analogues; signal transduction pathway inhibitors; non-receptor tyrosine kinase angiogenesis inhibitors; immunotherapeutic agents; pro-apoptotic agents; and cell cycle signaling inhibitors. 
     
     
         37 . The method of  claim 34 , wherein said chemotherapeutic agent is selected from the group consisting of taxol and taxotere. 
     
     
         38 . The method of  claim 32 , wherein said cancer is selected from the group consisting of carcinoma, including that of the bladder, breast, colon, head and neck, kidney, including renal cell carcinoma, liver, lung, ovary, pancreas, stomach, cervix, thyroid and skin; including squamous cell carcinoma; hematopoietic tumors of lymphoid lineage, including leukemia, acute lymphocytic leukemia, acute lymphoblastic leukemia, B-cell lymphoma, T-cell lymphoma, Burkitt's lymphoma; hematopoietic tumors of myeloid lineage, including acute and chronic myelogenous leukemias and promyelocytic leukemia; tumors of mesenchymal origin, including fibrosarcoma and rhabdomyoscarcoma; other tumors, including melanoma, seminoma, tetratocarcinoma, neuroblastoma and glioma; tumors of the central and peripheral nervous system, including astrocytoma, neuroblastoma, glioma, and schwannomas; tumors of mesenchymal origin, including fibrosarcoma, rhabdomyoscarama, and osteosarcoma; and other tumors, including melanoma, xeroderma pigmentosum, keratoactanthoma, seminoma, thyroid follicular cancer and teratocarcinoma. 
     
     
         39 . The method of  claim 38 , wherein said cancer is selected from the group consisting of melanoma, leukemia, renal cancer, colon cancer, ovarian cancer, prostate cancer, lung cancer, bladder cancer, breast cancer, and head and neck cancer