Active hedgehog protein conjugate

Abstract

A hedgehog conjugate which is characterized in that it contains: a) a polypeptide composed of 10 to 30 hydrophobic amino acids and/or amino acids which form transmembrane helices and are positively charged, b) 1 to 4 aliphatic, saturated or unsaturated hydrocarbon residues with a chain length of 10 to 24 C atoms and with a hydrophobic action or c) a hydrophobic thio compound covalently bound to a hedgehog protein and which has a several-fold increased activity and is suitable as a pharmaceutical agent.

1 . A hedgehog conjugate, comprising from 1 to 4 moieties covalently bound to a hedgehog protein; wherein each moiety is: 
 a) a polypeptide composed of 10 to 30 hydrophobic amino acids and/or of amino acids which form transmembrane helices and are positively charged,    b) an aliphatic, saturated or unsaturated hydrocarbon residues with a chain length of 8 to 24 C atoms and with a hydrophobic action, or    c) a hydrophobic thio group.    
     
     
         2 . The conjugate of  claim 1 , wherein each moiety is a polypeptide containing from 2 to 12 lysines and/or arginines.  
     
     
         3 . The conjugate of  claim 1 , wherein each moiety is 1 to 4 hydrocarbon residues and each hydrocarbon residue is a fatty acid or alkyl alcohol residue which is bound to the hedgehog protein by an ester, thioester, acid amide or disulfide bond.  
     
     
         4 . The conjugate of  claim 3 , wherein the fatty acid is lauric acid, myristic acid, palmitic acid, stearic acid, arachidic acid, behenic acid, palmitoleic acid, oleic acid, linoleic acid, linolenic acid or arachidonic acid.  
     
     
         5 . The conjugate of  claim 1 , wherein each moiety is the hydrophobic thio group thiocholesterol.  
     
     
         6 . The conjugate of  claim 1 , wherein the hydrocarbon residue, the polypeptide or the thio group is bound to the C-terminus and/or N-terminus of the hedgehog protein.  
     
     
         7 . A process for manufacturing the conjugate in  claim 3  comprising covalently coupling the hydrocarbon residue to a free hydroxy, mercapto, carboxy or amino group of the hedgehog protein.  
     
     
         8 . A process for manufacturing the conjugate of  claim 1  comprising covalently coupling a hydrocarbon residue or a hydrophobic thio compound to a hedgehog protein, wherein the covalent coupling of the hydrocarbon residue, the polypeptide or the hydrophobic thio compound and/or the isolation are carried out in the presence of suramin, heparin, anionic polysaccharides or a detergent.  
     
     
         9 . A process for manufacturing the conjugate of  claim 1 , comprising covalently coupling a fatty acid residue or a hydrophobic thio compound to a hedgehog protein, wherein the fatty acid residue is palmitoyl-CoA or palmitoylimidazolide and the hydrophobic thio group is thiocholesterol or a carbon chain containing a thiol group.  
     
     
         10 . A pharmaceutical composition containing the conjugate of  claim 1  in a pharmaceutically effective amount as well as an ingredient selected from the group consisting of pharmaceutical auxiliary substances, detergents, stabilizers matrix materials, suramin, heparin, anionic polysaccharides and sequestering agents.  
     
     
         11 . A hedgehog protein, wherein the thiol group of the N-terminal cysteine is coupled to a thiol-protecting group or said hedgehog protein is a homodimer wherein the N-terminal cysteines are linked via a disulfide bridge.  
     
     
         12 . A hedgehog conjugate comprising from 1 to 4 moieties covalently bound to a hedgehog protein wherein each moiety is: 
 a) a polypeptide composed of 10 to 30 hydrophobic amino acids and/or of amino acids which form transmembrane helices and are positively charged,    b) an aliphatic, saturated or unsaturated hydrocarbon residues with a chain length of 8 to 24 C atoms and with a hydrophobic action, or    c) a hydrophobic thio group.