Neurturin conjugates for pharmaceutical use
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. A neurturin conjugate comprising 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.
2. The neurturin conjugate according to claim 1 , wherein the polyol moiety is a polyethylene glycol moiety.
3. The neurturin conjugate according to claim 1 , wherein the polyol moiety is a single chain polyol moiety.
4. The neurturin conjugate according to claim 1 , wherein the polyol moiety is a branched chain polyol moiety.
5. The neurturin conjugate according to claim 1 , wherein the polyol moiety is a polyalkylene glycol moiety.
6. The neurturin conjugate according to claim 1 , wherein at least one of the arginine residues 51, 52, 54, 56, 57, 58, 60, 61, 63 or 65 is substituted with a lysine.
7. The neurturin conjugate of claim 6 , 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.
8. The neurturin conjugate of claim 1 , wherein the arginine residue 63 is substituted by lysine.
9. The neurturin conjugate of claim 1 , 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.
10. The neurturin conjugate of claim 1 , wherein said neurturin protein product has 1 amino acid alteration.
11. The neurturin conjugate of claim 1 , wherein said neurturin protein product has 2 amino acid alterations.
12. The neurturin conjugate of claim 1 , wherein said neurturin protein product has 3 amino acid alterations.
13. The neurturin conjugate of claim 1 , wherein said neurturin protein product has 4 amino acid alterations.
14. A pharmaceutical composition comprising at least one neurturin protein product 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 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.
15. The composition of claim 14 , wherein said neurturin protein product is a biologically active fragment of a neurturin protein having the amino acid sequence of SEQ ID NO: 5.
16. The composition of claim 14 , wherein said neurturin protein product is mono-PEGylated, comprising a single polyethylene glycol molecule chain.
17. The composition of claim 14 , wherein said neurturin protein product is oligo- or poly-PEGylated, comprising two, three, four or several polyethylene glycol molecule chains.
18. The composition of claim 14 , wherein said neurturin protein product comprises at least one linear polyethylene glycol molecule chain.
19. The composition of claim 14 , wherein said neurturin protein product comprises at least one branched polyethylene glycol molecule chain.
20. The composition of claim 14 , wherein said polyethylene glycol molecule has an average molecular weight of 100 to 10000 Da.
21. The composition of claim 14 , wherein said polyethylene glycol molecule is linked to the N-terminal amino acid of said neurturin protein product.
22. The composition of claim 14 , wherein said polyethylene glycol molecule is linked to said neurturin protein product via an acyl or alkyl linkage.
23. The composition of claim 14 , wherein said polyethylene glycol molecule is terminated by an OH—, OCH 3 — or OEt-group.
24. The composition of claim 14 , which has an increased bioavailability of the active ingredient compared to a composition with an unmodified control neurturin protein product.
25. The composition of claim 14 for injection or infusion.
26. The composition of claim 25 for subcutaneous or intravenous injection.
27. The composition of claim 14 for administration to a mammal.
28. The pharmaceutical composition of claim 14 , wherein said neurturin protein product has 1 amino acid alteration.
29. The pharmaceutical composition of claim 14 , wherein said neurturin protein product has 2 amino acid alterations.
30. The pharmaceutical composition of claim 14 , wherein said neurturin protein product has 3 amino acid alterations.
31. The pharmaceutical composition of claim 14 , wherein said neurturin protein product has 4 amino acid alterations.
32. A variant of human neurturin comprising a human neurturin protein product, wherein said human neurturin protein product is capable of dimerizing with a neurturin protein having the amino acid sequence of SEQ ID NO: 5, wherein said human 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.
33. The variant of human neurturin according to claim 32 , wherein the arginine residue at position 63 is substituted by lysine.
34. The variant of human neurturin according to claim 32 , wherein at least one arginine residue at amino acid position 51, 52, 54, 56, 57, 58, 60, 61 or 65 is substituted by glutamic acid.
35. The variant of human neurturin according to claim 32 , 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.
36. The human neurturin variant of claim 32 , wherein said neurturin protein product has 1 amino acid alteration.
37. The human neurturin variant of claim 32 , wherein said neurturin protein product has 2 amino acid alterations.
38. The human neurturin variant of claim 32 , wherein said neurturin protein product has 3 amino acid alterations.
39. The human neurturin variant of claim 32 , wherein said neurturin protein product has 4 amino acid alterations.