Agent for the treatment and/or prophylaxis of an autoimmune disease and for the formation of regulatory t cells

Abstract

The present invention relates to an agent for the treatment and/or prophylaxis of an autoimmune disease, an agent for the formation of regulatory T cells (T Reg ) in an organism and various methods in which the agents according to the invention are used.

1 . Method for the preparation of a medicament for the treatment and/or prophylaxis of an autoimmune disease, comprising the following steps:
 (1) providing a mutein of human interleukin-2 (hIL-2 mutein) or of a fragment thereof, which is numbered in accordance with the hIL-2 wild type and has an amino acid substitution in at least one of the positions 20, 88 or 126,   (2) formulating said mutein of fragment into a pharmaceutically acceptable carrier.   
     
     
         2 . Method according to  claim 1 , wherein through the substitution at position 88 an asparagine is exchanged for an amino acid which is selected from the group consisting of:
 arginine (hIL-2-N88R), glycine (hIL-2-N88G), or isoleucine (hIL-2-N88I).   
     
     
         3 . Method according to  claim 1 , wherein through the substitution at position 20 an aspartic acid is exchanged for an amino acid which is selected from the group consisting of: histidine (hIL-2-D2OH), isoleucine (hIL-2-D2OH), or tyrosine (hIL-2-D20Y). 
     
     
         4 . Method according to  claim 1 , wherein through the substitution at position 126, a glutamine is exchanged for a leucine (hIL-2-Q126L). 
     
     
         5 . Method according to  claim 1 , wherein the hIL-2 mutein or the fragment thereof has at least one further amino acid substitution in any position except the positions 20, 88 or 126, so that the thus further substituted hIL-2 mutein or the thus further substituted fragment thereof has an amino acid sequence which is at least 80% identical with the amino acid sequence of the hIL-2 mutein or of the section thereof, which is not further substituted, compared to the hIL-2 wild type, other than in at least one of the positions 20, 88 or 126. 
     
     
         6 . Method according to  claim 5 , wherein the further substitution in at least any position, except the positions 20, 88 or 126, is a conservative amino acid substitution. 
     
     
         7 . Method according to  claim 1 , wherein the medicament in addition contains an immunosuppressant. 
     
     
         8 . Method according to  claim 7 , wherein the immunosuppressant is selected from the group consisting of: glucocorticoid, including decortin, prednisol; azathioprine; cyclosporin A; mycophenolate mofetil; tacrolimus; anti-T lymphocyte globulin, anti-CD3 antibodies, including muromonab; anti-CD25 antibodies, including basiliximab and daclizumab; anti-TNF-α antibodies, including infliximab and adalimumab; azathioprine; methotrexate; cyclosporin; sirolimus; everolimus; fingolimod; CellCept®; myfortic; and cyclophosphamide. 
     
     
         9 . Method for the treatment and/or prophylaxis of an autoimmune disease in an organism, which comprises the following steps:
 (a) providing a mutein of human interleukin-2 (hIL-2 mutein) or of a fragment thereof, which is numbered in accordance with the hIL-2 wild type and has an amino acid substitution in at least one of the positions 20, 88 or 126,   (b) administering said hIL-2 mutein or of said fragment thereof to an organism, and   (c) if necessary repeating the steps (a) and (b).   
     
     
         10 . Method according to  claim 9 , wherein through the substitution at position 88 an asparagine is exchanged for an amino acid which is selected from the group consisting of: arginine (hIL-2-N88R), glycine (hIL-2-N88G), or isoleucine (hIL-2-N88I). 
     
     
         11 . Method according to  claim 9 , wherein through the substitution at position 20 an aspartic acid is exchanged for an amino acid which is selected from the group consisting of: histidine (hIL-2-D2OH), isoleucine (hIL-2-D2OH), or tyrosine (hIL-2-D20Y). 
     
     
         12 . Method according to  claim 9 , wherein through the substitution at position 126, a glutamine is exchanged for a leucine (hIL-2-Q126L). 
     
     
         13 . Method according to  claim 10 , wherein the hIL-2 mutein or the fragment thereof has at least one further amino acid substitution in any position except the positions 20, 88 or 126, so that the thus further substituted hIL-2 mutein or the thus further substituted fragment thereof has an amino acid sequence which is at least 80% identical with the amino acid sequence of the hIL-2 mutein or of the section thereof, which is not further substituted, compared to the hIL-2 wild type, other than in at least one of the positions 20, 88 or 126. 
     
     
         14 . Method according to  claim 13 , wherein the further substitution in at least any position, except the positions 20, 88 or 126, is a conservative amino acid substitution. 
     
     
         15 . Method according to  claim 9 , wherein the medicament in addition contains an immunosuppressant. 
     
     
         16 . Method according to  claim 15 , wherein the immunosuppressant is selected from the group consisting of: glucocorticoid, including decortin, prednisol; azathioprine; cyclosporin A; mycophenolate mofetil; tacrolimus; anti-T lymphocyte globulin, anti-CD3 antibodies, including muromonab; anti-CD25 antibodies, including basiliximab and daclizumab; anti-TNF-α antibodies, including infliximab and adalimumab; azathioprine; methotrexate; cyclosporin; sirolimus; everolimus; fingolimod; CellCept®; myfortic; and cyclophosphamide. 
     
     
         17 . Method for the treatment and/or prophylaxis of an autoimmune disease in an organism, which comprises the following steps:
 (a) providing a mutein of human interleukin-2 (hIL-2 mutein) or of a fragment thereof, which is numbered in accordance with the hIL-2 wild type and has an amino acid substitution in at least one of the positions 20, 88 or 126,   (b) contacting said hIL-2 mutein or of said fragment thereof with peripheral mononuclear blood cells (PBMCs) deriving from a first organism,   (c) incubating said hIL-2 mutein or of the fragment thereof with the PBMCs to obtain a cell population which contains regulatory T cells (T Reg ), and   (d) introducing the cell population into a second organism.   
     
     
         18 . Method according to  claim 17 , wherein the first and the second organisms are identical organisms.