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 - 38 . (canceled) 
     
     
         39 . A method of administering a neurturin conjugate to a subject having a pancreatic disorder, comprising administering to said subject a pharmaceutical composition comprising a neurturin conjugate and a pharmaceutically acceptable carrier, diluent and/or adjuvant, wherein said neurturin conjugate comprises a polyol moiety covalently bound to a human neurturin protein product, wherein said neurturin protein product is capable of dimerizing with a neurturin protein having the amino acid sequence of SEQ ID NO: 5, wherein said neurturin protein product comprises the amino acid sequence of SEQ ID NO: 5 or a biologically active fragment thereof, but with 1, 2, 3, or 4 amino acid alterations, wherein the amino acid alterations are: a) the substitution of 1, 2, 3 or 4 arginine residues at amino acid positions 51, 52, 54, 56, 57, 58, 60, 61, 63 or 65 of SEQ ID NO: 5 with a lysine, and/or b) the substitution of 1, 2, 3 or 4 arginine residues at amino acid positions 51, 52, 54, 56, 57, 58, 60, 61 or 65 of SEQ ID NO: 5 with a glutamic acid. 
     
     
         40 . The method of  claim 39 , wherein at least one of the amino acid alterations facilitates the conjugation of the polyol moiety covalently bound to the human neurturin protein product. 
     
     
         41 . The method of  claim 39 , wherein the amino acid alterations are the substitution of 1, 2, 3 or 4 arginine residues at amino acid positions 51, 52, 54, 56, 57, 58, 60, 61, 63 or 65 of SEQ ID NO: 5 with a lysine. 
     
     
         42 . The method of  claim 41 , wherein the neurturin conjugate 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. 
     
     
         43 . The method of  claim 39 , wherein the arginine residue 63 is substituted by lysine. 
     
     
         44 . The method of  claim 39 , wherein the amino acid alterations are the substitution of 1, 2, 3 or 4 arginine residues at amino acid positions 51, 52, 54, 56, 57, 58, 60, 61 or 65 of SEQ ID NO: 5 with a glutamic acid. 
     
     
         45 . The method of  claim 39 , wherein said neurturin protein product has 1 amino acid alteration. 
     
     
         46 . The method of  claim 39 , wherein said neurturin protein product has 2 amino acid alterations. 
     
     
         47 . The method of  claim 39 , wherein said neurturin protein product has 3 amino acid alterations. 
     
     
         48 . The method of  claim 39 , wherein said neurturin protein product has 4 amino acid alterations. 
     
     
         49 . The method of  claim 39 , wherein said neurturin protein product is a biologically active fragment of a neurturin protein having the amino acid sequence of SEQ ID NO: 5. 
     
     
         50 . The method according to  claim 39 , wherein the polyol moiety is a polyethylene glycol moiety. 
     
     
         51 . The method of  claim 39 , wherein the polyol moiety is a linear chain polyol moiety. 
     
     
         52 . The method of  claim 39 , wherein the polyol moiety is a branched chain polyol moiety. 
     
     
         53 . The method of  claim 39 , wherein the polyol moiety is a polyalkylene glycol moiety. 
     
     
         54 . The method of  claim 39 , wherein said neurturin protein product is mono-PEGylated, comprising a single polyethylene glycol molecule chain. 
     
     
         55 . The method of  claim 39 , wherein said neurturin protein product is oligo- or poly-PEGylated, comprising two, three, four or several polyethylene glycol molecule chains. 
     
     
         56 . The method of  claim 50 , wherein said polyethylene glycol molecule has an average molecular weight of 100 to 10000 Da. 
     
     
         57 . The method of  claim 50 , wherein said polyethylene glycol molecule is linked to the N-terminal amino acid of said neurturin protein product. 
     
     
         58 . The method of  claim 50 , wherein said polyethylene glycol molecule is linked to said neurturin protein product via an acyl or alkyl linkage. 
     
     
         59 . The method of  claim 50 , wherein said polyethylene glycol molecule is terminated by an OH—, OCH 3 — or OEt-group. 
     
     
         60 . The method of  claim 39 , wherein the neurturin conjugate has an increased bioavailability of the active ingredient compared to a composition with an unmodified control neurturin protein product. 
     
     
         61 . The method of  claim 39 , wherein the composition is administered to the subject by injection or infusion. 
     
     
         62 . The method of  claim 61 , wherein the composition is administered to the subject by subcutaneous or intravenous injection. 
     
     
         63 . The method of  claim 39 , wherein the subject is a mammal. 
     
     
         64 . The method of  claim 39 , wherein the pancreatic disorder is Type I diabetes. 
     
     
         65 . The method of  claim 39 , wherein the pancreatic disorder is Type II diabetes. 
     
     
         66 . The method of  claim 39 , wherein the pancreatic disorder is latent autoimmune diabetes in adults (LADA).