Biologically active complex of NR6 and cardiotrophin-like-cytokine

Abstract

The present invention relates generally to a biologically active complex comprising at least two heterologous molecules. More particularly, the biologically active complex of the present invention comprises at least two polypeptides or parts, fragments, truncates or protease-activated forms of one or more of the polypeptides wherein the complex alone or in association with a receptor, ligand or other molecule facilitates proliferation, differentiation and/or survival of a cell. The identification of the biologically active complex of the present invention permits the assay for agonists and antagonists of the formation of the biologically active complex as well as therapeutic and diagnostic reagents based on the biologically active complex or interaction between the biologically active complex and a receptor, ligand or other molecule.

1 . A biologically active complex comprising at least two heterologous molecules, which complex alone or in association with a receptor, ligand or other molecule facilitates proliferation, differentiation and/or survival of a cell.  
     
     
         2 . A biologically active complex according to  claim 1  wherein the heterologous molecules are peptides, polypeptides or proteins or parts, fragments, truncates or protease-activated forms thereof.  
     
     
         3 . A biologically active complex according to  claim 2  wherein the two peptides, polypeptides or proteins are encoded by two different genes or are encoded by splice variants of the same gene.  
     
     
         4 . A biologically active complex according to  claim 3  wherein at least one polypeptide or protein within the complex is a soluble haemopoietin receptor.  
     
     
         5 . A biologically active complex according to  claim 4  wherein the at least one polypeptide or protein is the haemopoietin receptor, NR6.  
     
     
         6 . A biologically active complex according to  claim 4  or  5  wherein the other of said polypeptide or protein is a cytokine or cytokine-like molecule.  
     
     
         7 . A biologically active complex according to  claim 6  wherein said other polypeptide is cardiotrophin-like cytokine (CLC) or a part, fragment, truncate or protease-activated form thereof.  
     
     
         8 . A biologically active complex according to  claim 2  comprising the structure:  
         [X 1 ] n  (a) [X 2 ] n1  (b) [X 3 ] n2  . . . [X d ] n3    
       wherein 
 X 1  and X 2  are different and one is NR6 and the other is CLC or parts, fragments, truncates or protease-activated forms thereof;  
 X 3  . . . X d  are optionally present represent other members of the complex such as a cytokine or cytokine-like molecule;  
 n and n 1  may be the same or different and each is from about 1 to about 50;  
 n 2  and n 3  may be the same or different and each is from 0 to about 50;  
 (a) and (b) may be the same or different and represent the bonds, interactions or other “forces” which keep the members together in the complex.  
 
     
     
         9 . A biologically active complex according to  claim 8  wherein X 3  is selected from CNTFR, gp130, LIFRα or other receptor molecule or cytokine-like molecule.  
     
     
         10 . A biologically active molecule according to  claim 2  comprising the structure:  
         [X 1 ] a3  [NR6] a  [CLC] a1  [NR6] a2  [X 1 ] a4    
       wherein: 
 X 1  is optionally present and is a cytokine or cytokine-like molecule;  
 a is from about 0 to 10;  
 a 1  is from 1 to about 10;  
 a 2  is from 0 to 10;  
 with the proviso that if one of a or a 2  is 0 then the other of a or a 2  cannot be 0;  
 a 3  is from about 0 to 10;  
 a 4  is from about 0 to 10;  
 with the proviso that if X 1  is present then either a 3  or a 4  is 0.  
 
     
     
         11 . A biologically active complex according to any one of  claims 5  to  10  wherein the NR6 and/or CLC is of animal or avian origin.  
     
     
         12 . A biologically active complex according to  claim 11  wherein the NR6 and/or CLC is of human, primate or murine origin.  
     
     
         13 . A genetic construct comprising a nucleotide sequence encoding a biologically active complex according to  claim 2 .  
     
     
         14 . A genetic construct according to  claim 13  comprising a first nucleotide sequence encoding one or other of NR6 or CLC or modified forms thereof and a second nucleotide sequence encoding the other of NR6 or CLC.  
     
     
         15 . A genetic construct according to  claim 14  comprising a sequence of nucleotides substantially as set forth in <400>1 or a nucleotide sequence having at least 60% similarity to the nucleotide sequence set forth in <400>1 or a nucleotide sequence capable of hybridizing thereto under low stringency conditions at 42° C. and a sequence of nucleotides encoding CLC or a modified form thereof.  
     
     
         16 . A genetic construct according to  claim 14  comprising a sequence of nucleotides substantially as set forth in <400>3 or a nucleotide sequence having at least 60% similarity to the nucleotide sequence set forth in <400>3 or a nucleotide sequence capable of hybridizing thereto under low stringency conditions at 42° C. and a sequence of nucleotides encoding CLC or a modified form thereof.  
     
     
         17 . A genetic construct according to  claim 14  comprising a sequence of nucleotides substantially as set forth in <400>5 or a nucleotide sequence having at least 60% similarity to the nucleotide sequence set forth in <400>5 or a nucleotide sequence capable of hybridizing thereto under low stringency conditions at 42° C. and a sequence of nucleotides encoding CLC or a modified form thereof.  
     
     
         18 . A genetic construct according to  claim 14  comprising a sequence of nucleotides substantially as set forth in <400>7 or a nucleotide sequence having at least 60% similarity to the nucleotide sequence set forth in <400>7 or a nucleotide sequence capable of hybridizing thereto under low stringency conditions at 42° C. and a sequence of nucleotides encoding CLC or a modified form thereof.  
     
     
         19 . A genetic construct according to  claim 14  comprising a sequence of nucleotides substantially as set forth in <400>9 or a nucleotide sequence having at least 60% similarity to the nucleotide sequence set forth in <400>9 or a nucleotide sequence capable of hybridizing thereto under low stringency conditions at 42° C. and a sequence of nucleotides encoding CLC or a modified form thereof.  
     
     
         20 . A genetic construct according to  claim 14  comprising a sequence of nucleotides substantially as set forth in <400>11 or a nucleotide sequence having at least 60% similarity to the nucleotide sequence set forth in <400>11 or a nucleotide sequence capable of hybridizing thereto under low stringency conditions at 42° C. and a sequence of nucleotides encoding CLC or a modified form thereof.  
     
     
         21 . A genetic construct according to  claim 14  comprising a sequence of nucleotides substantially as set forth in <400>12 or a nucleotide sequence having at least 60% similarity to the nucleotide sequence set forth in <400>12 or a nucleotide sequence capable of hybridizing thereto under low stringency conditions at 42° C. and a sequence of nucleotides encoding CLC or a modified form thereof.  
     
     
         22 . A genetic construct according to  claim 14  comprising a sequence of nucleotides substantially as set forth in <400>13 or a nucleotide sequence having at least 60% similarity to the nucleotide sequence set forth in <400>13 or a nucleotide sequence capable of hybridizing thereto under low stringency conditions at 42° C. and a sequence of nucleotides encoding CLC or a modified form thereof.  
     
     
         23 . An expression vector encoding a biologically active complex according to  claim 2 .  
     
     
         24 . An expression vector according to  claim 23  comprising a nucleic acid molecule encoding NR6 and CLC or modified forms thereof, said expression vector capable of expression in a selected host cell.  
     
     
         25 . An expression vector according to  claim 23  comprising a sequence of nucleotides encoding NR6 or a derivative thereof having an amino acid sequence as set forth in <400>2 or having at least about 50% similarity to all or part thereof, said genetic construct further comprising a sequence of nucleotides encoding CLC or a modified form thereof.  
     
     
         26 . An expression vector according to  claim 23  comprising a sequence of nucleotides encoding NR6 or a derivative thereof having an amino acid sequence as set forth in <400>4 or having at least about 50% similarity to all or part thereof, said genetic construct further comprising a sequence of nucleotides encoding CLC or a modified form thereof.  
     
     
         27 . An expression vector according to  claim 23  comprising a sequence of nucleotides encoding NR6 or a derivative thereof having an amino acid sequence as set forth in <400>6 or having at least about 50% similarity to all or part thereof, said genetic construct further comprising a sequence of nucleotides encoding CLC or a modified form thereof.  
     
     
         28 . An expression vector according to  claim 23  comprising a sequence of nucleotides encoding NR6 or a derivative thereof having an amino acid sequence as set forth in <400>8 or having at least about 50% similarity to all or part thereof, said genetic construct further comprising a sequence of nucleotides encoding CLC or a modified form thereof.  
     
     
         29 . An expression vector according to  claim 23  comprising a sequence of nucleotides encoding NR6 or a derivative thereof having an amino acid sequence as set forth in <400>14 or having at least about 50% similarity to all or part thereof, said genetic construct further comprising a sequence of nucleotides encoding CLC or a modified form thereof.  
     
     
         30 . An expression vector according to  claim 23  comprising a sequence of nucleotides encoding NR6 or a derivative thereof having an amino acid sequence as set forth in <400>15 or having at least about 50% similarity to all or part thereof, said genetic construct further comprising a sequence of nucleotides encoding CLC or a modified form thereof.  
     
     
         31 . An expression vector according to  claim 23  comprising a sequence of nucleotides encoding NR6 or a derivative thereof having an amino acid sequence as set forth in <400>19 or having at least about 50% similarity to all or part thereof, said genetic construct further comprising a sequence of nucleotides encoding CLC or a modified form thereof.  
     
     
         32 . An expression vector according to  claim 23  comprising a sequence of nucleotides encoding NR6 or a derivative thereof having an amino acid sequence as set forth in <400>20 or having at least about 50% similarity to all or part thereof, said genetic construct further comprising a sequence of nucleotides encoding CLC or a modified form thereof.  
     
     
         33 . A method of modulating the activity of a biological complex according to any one of  claims 1  to  12  in a subject, said method comprising administering to said subject a modulating effective amount of a molecule for a time and under conditions sufficient to increase or decrease the biological activity of the complex.  
     
     
         34 . A composition comprising a biologically active complex according to any one of  claims 1  to  12  and one or more pharmaceutically active carriers and/or diluents.  
     
     
         35 . A method of treatment or prophylaxis of a subject, said method comprising administering to said subject a biologically active complex comprising at least two heterologous molecules which complex alone or in association with a receptor, ligand or other molecule facilitates proliferation, differentiation and/or survival of a cell.  
     
     
         36 . A method according to  claim 35  wherein the heterologous molecules are peptides, polypeptides or proteins or parts, fragments, truncates or protease-activated forms thereof.  
     
     
         37 . A method according to  claim 36  wherein the two peptides, polypeptides or proteins are encoded by two different genes or are encoded by splice variants of the same gene.  
     
     
         38 . A method according to  claim 36  wherein at least one polypeptide or protein within the complex is a soluble haemopoietin receptor.  
     
     
         39 . A method according to  claim 38  wherein the at least one polypeptide or protein is the haemopoietin receptor, NR6.  
     
     
         40 . A method according to  claim 38  or  39  wherein the other of said polypeptide or protein is a cytokine or cytokine-like molecule.  
     
     
         41 . A method according to  claim 40  wherein said other polypeptide is a cardiotrophin-like cytokine (CLC) or a part, fragment, truncate or protease-activated form thereof.  
     
     
         42 . A method according to  claim 35  comprising the structure:  
         [X 1 ] a3  [NR6] a  [CLC] a1  [NR6] a2  [X 1 ] a4    
       wherein: 
 X 1  is optionally present and is a cytokine or cytokine-like molecule;  
 a is from about 0 to 10;  
 a 1  is from 1 to about 10;  
 a 2  is from 0 to 10;  
 with the proviso that if one of a or a 2  is 0 then the other of a or a 2  cannot be 0;  
 a 3  is from about 0 to 10;  
 a 4  is from about 0 to 10;  
 with the proviso that if X 1  is present then either a 3  or a 4  is 0.  
 
     
     
         43 . A method according to  claim 42  wherein X 3  is selected from CNTFR, gp130, LIFRα or other receptor molecule or cytokine-like molecule.  
     
     
         44 . A method according to  claim 35  comprising the structure:  
         [X 1 ] a3  [NR6] a  [CLC] n1  [NR6] a2  [X 1 ] a4    
       wherein: 
 X 1  is optionally present and is a cytokine or cytokine-like molecule;  
 a is from about 0 to 10;  
 a 1  is from 1 to about 10;  
 a 2  is from 0 to 10;  
 with the proviso that if one of a or a 2  is 0 then the other of a or a 2  cannot be 0;  
 a 3  is from about 0 to 10;  
 a 4  is from about 0 to 10;  
 with the proviso that if X 1  is present then either a 3  or a 4  is 0.  
 
     
     
         45 . A method according to any one of  claims 39  to  44  wherein the NR6 and/or CLC is of animal or avian origin.  
     
     
         46 . A method according to  claim 45  wherein the NR6 and/or CLC is of human, primate or murine origin.  
     
     
         47 . Use of a biologically active complex according to any one of  claims 1  to  12  in the manufacture of a medicament for the treatment and/or prophylaxis of a disease condition in a subject.  
     
     
         48 . An antibody to a biologically active complex according to any one of  claims 1  to  12  or an antigenic binding portion of said antibody.  
     
     
         49 . An antibody according to  claim 48  where the antibody is a monoclonal antibody.  
     
     
         50 . A method of identifying an agent capable of modulating the effects of a biologically active complex according to any one of  claims 1  to  12 , said method comprising screening for agents which are capable of interacting with the complex or interfering or otherwise antagonizing or promoting or otherwise agonizing interaction between the heterologous molecules of said complex.  
     
     
         51 . An agent useful for modulating the effects of a biologically active complex according to any one of  claims 1  to  12  wherein said agent is capable of interacting with the complex or interfering or otherwise antagonizing or promoting or otherwise agonizing interaction between the heterologous molecules of said complex.