Chimeric proteins with cell-targeting specificity and apoptosis-inducing activities

Abstract

The present invention relates to chimeric proteins with cell-targeting specificity and apoptosis-inducing activities. In particular, the invention is illustrated by a recombinant chimeric protein between human interleukin-2 (IL2) and Bax. The chimeric protein specifically targets IL2 receptor (IL2R)-expressing cells and induces cell-specific apoptosis.

1 . A chimeric protein comprising a cell-specific targeting moiety and an apoptosis-inducing moiety, wherein the chimeric protein is produced by a recombinant DNA method, the targeting moiety is selected from the group consisting of a cytokine, a growth factor, a hormone, an antibody and a binding fragment of an antibody, and the apoptosis-inducing moiety is a caspase or a caspase subunit that induces apoptosis, cytochrome C, a DNA fragmentation factor (DFF), a domain of an apoptotic protein of the Bcl-2 family which domain has apoptosis-inducing activity, or a domain of DFF which domain has apoptosis-inducing activity. 
     
     
         2 . The chimeric protein of  claim 1  in which the apoptosis-inducing moiety is a protein of human origin. 
     
     
         3 . The chimeric protein of  claim 1  in which the apoptosis-inducing moiety is DFF 40. 
     
     
         4 . The chimeric protein of  claim 1  in which the apoptosis-inducing moiety is a caspase. 
     
     
         5 . The chimeric protein of  claim 1  in which the apoptosis-inducing moiety is caspase 3. 
     
     
         6 . The chimeric protein of  claim 1  in which the apoptosis-inducing moiety is cytochrome C. 
     
     
         7 . The chimeric protein of  claim 1  in which the apoptosis-inducing moiety is a fragment of an apoptotic protein of the Bcl-2 family. 
     
     
         8 . The chimeric protein of  claim 1  in which the cell-specific targeting moiety binds interleukin 2 receptor-expressing cells. 
     
     
         9 . The chimeric protein of  claim 1  in which the cell-specific targeting moiety is an interleukin. 
     
     
         10 . The chimeric protein of  claim 9  in which the cell-specific targeting moiety is an interleukin 2. 
     
     
         11 . The chimeric protein of  claim 1  in which the cell-specific targeting moiety is myelin basic protein. 
     
     
         12 . The chimeric protein of  claim 1  in which the cell-specific targeting moiety binds tumor cells. 
     
     
         13 . The chimeric protein of  claim 1  in which the cell-specific targeting moiety is an antibody or a fragment thereof. 
     
     
         14 . The chimeric protein of  claim 13  in which the cell-specific targeting moiety is a single chain antibody. 
     
     
         15 . The chimeric protein of  claim 13  in which the cell-specific targeting moiety is a Fc fragment of an IgE antibody. 
     
     
         16 . The chimeric protein of  claim 1  in which the cell-specific targeting moiety is a cytokine. 
     
     
         17 . The chimeric protein of  claim 1  in which the cell-specific targeting moiety is epidermal growth factor. 
     
     
         18 . The chimeric protein of  claim 1  in which the cell-specific targeting moiety is insulin-like growth factor. 
     
     
         19 . The chimeric protein of  claim 1  in which the two moieties are connected by a polylinker. 
     
     
         20 . A pharmaceutical composition comprising a chimeric protein according to  claim 1  and a pharmaceutically acceptable carrier. 
     
     
         21 . A pharmaceutical composition comprising a chimeric protein according to  claim 2  and a pharmaceutically acceptable carrier. 
     
     
         22 . A pharmaceutical composition comprising a chimeric protein according to  claim 5  and a pharmaceutically acceptable carrier. 
     
     
         23 . A pharmaceutical composition comprising a chimeric protein according to  claim 7  and a pharmaceutically acceptable carrier. 
     
     
         24 . A pharmaceutical composition comprising a chimeric protein according to  claim 8  and a pharmaceutically acceptable carrier. 
     
     
         25 . A pharmaceutical composition comprising a chimeric protein according to  claim 11  and a pharmaceutically acceptable carrier. 
     
     
         26 . A pharmaceutical composition comprising a chimeric protein according to  claim 12  and a pharmaceutically acceptable carrier. 
     
     
         27 . A pharmaceutical composition comprising a chimeric protein according to  claim 13 - 15  and a pharmaceutically acceptable carrier. 
     
     
         28 . A pharmaceutical composition comprising a chimeric protein according to  claim 16  and a pharmaceutically acceptable carrier. 
     
     
         29 . A pharmaceutical composition comprising a chimeric protein according to  claim 19  and a pharmaceutically acceptable carrier. 
     
     
         30 . A method of treating a pathological disorder associated with undesirable target cells, comprising administering to a patient in need thereof a chimeric protein comprising a targeting moiety specific for the undesirable cells and an apoptosis-inducing moiety, wherein the apoptosis-inducing moiety is a caspase, cytochrome C, a DNA fragmentation factor (DFF), a fragment of an apoptotic protein of the Bcl-2 family which fragment has apoptosis-inducing activity, or a domain of caspase, cytochrome C or DFF which domain has apoptosis-inducing activity. 
     
     
         31 . A method of treating a malignant or pre-malignant condition, comprising administering to a patient in need thereof a chimeric protein of  claim 1  or a pharmaceutical composition of  claim 20 . 
     
     
         32 . A method of treating an immune disorder associated with cells expressing and IL-2 receptor comprising administering to a patient in need thereof a chimeric protein of  claim 10 . 
     
     
         33 . A method of treating a hypersensitivity condition, comprising administering to a patient in need thereof a chimeric protein of  claim 1  or a pharmaceutical composition of  claim 20 . 
     
     
         34 . A method of treating an infectious disease comprising administering to a patient in need thereof a chimeric protein of  claim 1  or a pharmaceutical composition of  claim 20 . 
     
     
         35 . A pharmaceutical composition comprising a chimeric protein according to  claim 4  and a pharmaceutically acceptable carrier. 
     
     
         36 . A pharmaceutical composition comprising a chimeric protein according to  claim 9  and a pharmaceutically acceptable carrier. 
     
     
         37 . A pharmaceutical composition comprising a chimeric protein according to  claim 10  and a pharmaceutically acceptable carrier. 
     
     
         38 . The chimeric protein of  claim 1  wherein the apoptosis-inducing moiety is the domain of an apoptotic protein of the Bcl-2 family which domain has apoptosis-inducing activity and the domain is a BH3 domain. 
     
     
         39 . A chimeric protein comprising a cell-specific targeting moiety and an apoptosis-inducing moiety, wherein the chimeric protein is produced by a recombinant DNA method, the targeting moiety is selected from the group consisting of a cytokine, a growth factor, a hormone, an antibody and a binding fragment of an antibody, and the apoptosis-inducing moiety is a caspase or a caspase subunit that induces apoptosis. 
     
     
         40 . The Chimeric protein of  claim 39  wherein the targeting moiety is an IL-2.