Novel neurturin conjugates for pharmaceutical use

Abstract

The present invention relates to neurturin protein products conjugated to polyols and to pharmaceutical compositions comprising neurturin conjugates as active ingredients, preferably PEGylated neurturin conjugates or variants thereof, having increased bioavailability.

1 - 2 . (canceled) 
     
     
         3 . The neurturin conjugate according to  claim 9 , wherein the polyol moiety is a polyethylene glycol moiety. 
     
     
         4 . The neurturin conjugate according to  claim 9 , wherein the polyol moiety is a single chain polyol moiety. 
     
     
         5 . The neurturin conjugate according to  claim 9 , wherein the polyol moiety is a branched chain polyol moiety. 
     
     
         6 . The neurturin conjugate according to  claim 9 , wherein the polyol moiety is a polyalkylene glycol moiety. 
     
     
         7 . The neurturin conjugate according to  claim 9 , having the same or a higher in vivo neurturin activity as native human neurturin. 
     
     
         8 . (canceled) 
     
     
         9 . A neurturin conjugate comprising a polyol moiety covalently bound to a human neurturin protein product or a biologically active fragment thereof, wherein said neurturin protein product has an amino acid sequence that is at least 90% identical to SEQ ID NO: 5, wherein said neurturin protein product or biologically active fragment thereof is capable of dimerizing with a neurturin protein having the amino acid sequence of SEQ ID NO: 5, and wherein said neurturin protein product or biologically active fragment thereof has:
 a) at least one of the arginine residues corresponding to amino acid positions 51, 52, 54, 56, 57, 58, 60, 61, 63 or 65 of SEQ ID NO: 5 substituted with a lysine, and/or   b) at least one of the arginine residues corresponding to amino acid positions 51, 52, 54, 56, 57, 58, 60, 61 or 65 of SEQ ID NO: 5 substituted with a neutral or acidic amino acid.   
     
     
         10 . The neurturin conjugate according to  claim 9 , wherein at least one of the arginine residues 51, 52, 54, 56, 57, 58, 60, 61, 63 or 65 is substituted with a lysine. 
     
     
         11 . The neurturin conjugate according to  claim 9 , wherein at least one of the arginine residues 51, 52, 54, 56, 57, 58, 60, 61, or 65 is substituted by a neutral or acidic amino acid residue. 
     
     
         12 - 13 . (canceled) 
     
     
         14 . The neurturin conjugate of  claim 9 , wherein the arginine residue 63 is substituted by lysine. 
     
     
         15 . The neurturin conjugate of  claim 10 , which consists of two neurturin monomers, and which comprises 1-4 polyol moieties bound to the N-terminus and/or bound to a lysine side chain. 
     
     
         16 . A pharmaceutical composition comprising at least one neurturin protein product and/or a biologically active fragment thereof conjugated to at least one polyethylene glycol molecule as an active ingredient together with a pharmaceutically acceptable carrier, diluent and/or adjuvant, wherein said neurturin protein product has an amino acid sequence that is at least 90% identical to SEQ ID NO: 5, wherein said neurturin protein product or biologically active fragment thereof is capable of dimerizing with a neurturin protein having the amino acid sequence of SEQ ID NO: 5, and wherein said neurturin protein product or biologically active fragment thereof has:
 a) at least one of the arginine residues 51, 52, 54, 56, 57, 58, 60, 61, 63 or 65 substituted with a lysine, and/or   b) at least one of the arginine residues 51, 52, 54, 56, 57, 58, 60, 61, or 65 substituted with a neutral or acidic amino acid.   
     
     
         17 . The composition of  claim 16 , wherein said neurturin protein product is a biologically active fragment thereof. 
     
     
         18 . The composition of  claim 16 , wherein said neurturin protein product is mono-PEGylated, comprising a single polyethylene glycol molecule chain. 
     
     
         19 . The composition of  claim 16 , wherein said neurturin protein product is oligo- or poly-PEGylated, comprising two, three, four or several polyethylene glycol molecule chains. 
     
     
         20 . The composition of  claim 16 , wherein said neurturin protein product comprises at least one linear polyethylene glycol molecule chain. 
     
     
         21 . The composition of  claim 16 , wherein said neurturin protein product comprises at least one branched polyethylene glycol molecule chain. 
     
     
         22 . The composition of  claim 16 , wherein said polyethylene glycol molecule has an average molecular weight of 100 to 10000 Da. 
     
     
         23 . The composition of  claim 16 , wherein said polyethylene glycol molecule is linked to the N-terminal amino acid of said neurturin protein product. 
     
     
         24 . The composition of  claim 16 , wherein said polyethylene glycol molecule is linked to said neurturin protein product via an acyl or alkyl linkage. 
     
     
         25 . The composition of  claim 16 , wherein said polyethylene glycol molecule is terminated by an OH-, OCH 3 - or OEt-group. 
     
     
         26 . The composition of  claim 16 , which has an increased bioavailability of the active ingredient compared to a composition with an unmodified control neurturin protein product. 
     
     
         27 . The composition of  claim 16 , wherein said neurturin protein is present in an amount to provide an at least 2-fold increase in the bioavailability of the active ingredient. 
     
     
         28 - 31 . (canceled) 
     
     
         32 . The composition of  claim 16  for administration to a mammal. 
     
     
         33 . The variant of human neurturin of  claim 34 , wherein said neurturin protein product comprises 2, 3 or 4 amino acid changes compared to wild-type human neurturin. 
     
     
         34 . A variant of human neurturin comprising a human neurturin protein product or a biologically active fragment thereof, wherein said human neurturin protein product has an amino acid sequence that is at least 90% identical to SEQ ID NO: 5, wherein said neurturin protein product or biologically active fragment thereof is capable of dimerizing with a neurturin protein having the amino acid sequence of SEQ ID NO: 5, and wherein said neurturin protein product or biologically active fragment thereof has:
 a) at least one of the arginine residues 51, 52, 54, 56, 57, 58, 60, 61, 63 or 65 substituted with a lysine, and/or   b) at least one of the arginine residues 51, 52, 54, 56, 57, 58, 60, 61, or 65 substituted with a neutral or acidic amino acid.   
     
     
         35 . (canceled) 
     
     
         36 . A method for administering a pharmaceutical composition to a subject in need thereof comprising a pharmaceutically active amount of the neurturin protein product or a biologically active fragment thereof of  claim 9 , wherein said neurturin protein product or biologically active fragment thereof is conjugated to at least one polyethylene glycol molecule as an active ingredient together with a pharmaceutically acceptable carrier, diluent and/or adjuvant. 
     
     
         37 . The method of  claim 36 , wherein the subject is human. 
     
     
         38 . (canceled) 
     
     
         39 . The variant of human neurturin according to  claim 34 , wherein the arginine residue at position 63 is substituted by lysine. 
     
     
         40 . The variant of human neurturin according to  claim 34 , wherein at least one arginine residue 51, 52, 54, 56, 57, 58, 60, 61, or 65 is substituted by glutamic acid. 
     
     
         41 . The variant of human neurturin according to  claim 34 , wherein said at least one amino acid change is a substitution of at least one arginine residue with a neutral or acidic amino acid residue.