Interleukin-4 receptor-binding fusion proteins and uses thereof

Abstract

The present invention relates to interleukin-4 receptor binding fusion proteins. More specifically, the invention provides, in part, fusion proteins that include an interleukin-4 receptor binding protein moiety joined to a pro-apoptotic Bcl-2 family member protein moiety.

What is claimed is: 
     
         1 . A fusion protein comprising an interleukin-4 (1L-4) receptor binding protein and a pro-apoptotic Bcl-2 family polypeptide 
     
     
         2 . The fusion protein of  claim 1  wherein the IL-4 receptor binding protein is circularly permuted (cp). 
     
     
         3 . The fusion protein of  claim 1  or  2  wherein the pro-apoptotic Bcl-2 family polypeptide comprises a BH3 domain. 
     
     
         4 . The fusion protein of  claim 3  wherein the pro-apoptotic Bcl-2 family polypeptide comprising a BH3 domain is Bad, Bik/Nbk, Bid, Bim/Bod, Hrk, Bak or Bax. 
     
     
         5 . The fusion protein of  claim 3  or  4  wherein the pro-apoptotic Bcl-2 family polypeptide comprising a BH3 domain further comprises a mutation that reduces phosphorylation. 
     
     
         6 . The fusion protein of  claim 5  wherein the pro-apoptotic Bel-2 family polypeptide comprising a BH3 domain that further comprises a mutation that reduces phosphorylation is a Bad polypeptide. 
     
     
         7 . The fusion protein of any one of  claims 1  to  6 , wherein the fusion protein is capable of inhibiting cell survival, inhibiting cell proliferation, or enhancing cell death or apoptosis of a target cell expressing an IL-4 receptor (IL-4R). 
     
     
         8 . The fusion protein of any one of  claims 1  to  7  wherein the IL-4 receptor binding protein is a mutant IL-4 or IL-13 selective for binding to a Type I or a Type II IL-4R. 
     
     
         9 . The fusion protein of  claim 8  wherein the mutant IL-4 selective for binding to a Type II IL-4R comprises a KFR variant or a KF variant or the mutant IL-4 selective for binding to a Type I IL-4R comprises an RGA variant. 
     
     
         10 . The fusion protein of  claim 8  wherein the mutant IL-13 comprises an All variant or a DN variant. 
     
     
         11 . The fusion protein of any one of  claims 1  to  10  further comprising a linker. 
     
     
         12 . The fusion protein of  claim 11  wherein the linker has the sequence GS or is a ubiquitin or ubiquitin variant molecule. 
     
     
         13 . The fusion protein of  claim 1  or  2 , comprising the amino acid sequence of any one of SEQ ID NOs: 24-27. 
     
     
         14 . A nucleic acid molecule encoding the fusion protein of any one of  claims 1  to  13 . 
     
     
         15 . A nucleic acid molecule comprising the nucleic acid sequence of any one of SEQ ID NOs: 35-38. 
     
     
         16 . A vector comprising the nucleic acid molecule of  claim 14  or  15 . 
     
     
         17 . A host cell comprising the vector of  claim 16 . 
     
     
         18 . A pharmaceutical composition comprising the fusion protein of any one of  claims 1  to  13 , the nucleic acid molecule of  claim 14  or  15 , the vector of  claim 16 , or the host cell of  claim 17 . 
     
     
         19 . A method of inducing cell death comprising administering: the fusion protein of any one of  claims 1  to  13 , the nucleic acid molecule of  claim 14  or  15 , the vector of  claim 16 , or the host cell of  claim 17 , to a subject in need thereof. 
     
     
         20 . A method of inducing cell death comprising contacting a target cell that expresses an IL-4R with the fusion protein of any one of  claims 1  to  13 , the nucleic acid molecule of  claim 14  or  15 , or the vector of  claim 16 . 
     
     
         21 . A method of treating cancer comprising administering: the fusion protein of any one of  claims 1  to  13 , the nucleic acid molecule of  claim 14  or  15 , the vector of  claim 16 , or the host cell of  claim 17 , to a subject in need thereof 
     
     
         22 . A method of treating cancer comprising contacting a neoplastic cell that expresses an IL-4R with the fusion protein of any one of  claims 1  to  13 , the nucleic acid molecule of  claim 14  or  15 , or the vector of  claim 16 . 
     
     
         23 . A method of treating cancer comprising contacting a non-malignant cell that expresses an IL-4R in a tumour microenvironment in a subject in need thereof with the fusion protein of any one of  claims 1  to  13 , the nucleic acid molecule of  claim 14  or  15 , the vector of  claim 16 , or the host cell of  claim 17 . 
     
     
         24 . The method of  claim 23  wherein the non-malignant cell is contacted prior to said subject starting a therapy. 
     
     
         25 . A method of treating a hyperproliferative or differentiative disorder comprising administering: the fusion protein of any one of  claims 1  to  13 , the nucleic acid molecule of  claim 14  or  15 , the vector of  claim 16 , or the host cell of  claim 17 , to a subject in need thereof. 
     
     
         26 . The method of  claim 25  wherein the hyperproliferative or differentiative disorder is a fibrosis or hyperplasia, an inflammatory conditions or an autoimmune condition. 
     
     
         27 . The method of  claim 26  wherein the fibrosis or hyperplasia is pulmonary fibrosis or hyperplasia (such as benign prostatic hyperplasia), cardiac fibrosis, or liver fibrosis; the inflammatory condition is prostatitis, vernal keratoconjunctivitis, artherosclerosis, or idiopathic pulmonary pneumonia; and the autoimmune condition is Graves disease. 
     
     
         28 . Use of the fusion protein of any one of  claims 1  to  13 , the nucleic acid molecule of  claim 14  or  15 , the vector of  claim 16 , or the host cell of  claim 17 , for inducing cell death or treating cancer or treating a hyperproliferative or differentiative disorder in a subject in need thereof. 
     
     
         29 . The method of  claim 19 ,  21 ,  23 - 27  or the use of  claim 28 , wherein the subject is a human.