Claims
- 1. A GLP-2 derivative comprising a lipophilic substituent attached to any one amino acid residue.
- 2. A GLP-2 derivative of claim 1 with the proviso that only if the substituent has an ω-carboxylic acid group or is an alkyl group can it be attached to the N-terminal or C-terminal amino acid residue of the parent peptide.
- 3. A GLP-2 derivative of claim 1 or 2, wherein the lipophilic substituent comprises from 4 to 40 carbon atoms, more preferred from 8 to 25.
- 4. A GLP-2 derivative of any of the preceding claims, wherein said lipophilic substituent is attached to said amino acid in such a way that a carboxyl group of the lipophilic substituent forms an amide bond with an amino group of the amino acid.
- 5. A GLP-2 derivative of any of claims 1-3, wherein said lipophilic substituent is attached to said amino acid in such a way that an amino group of the lipophilic substituent forms an amide bond with a carboxyl group of the amino acid.
- 6. A GLP-2 derivative of any of the preceding claims, wherein the lipophilic substituent is attached to the parent peptide by means of a spacer.
- 7. A GLP-2 derivative of claim 6, wherein the spacer is an unbranched alkane α,ω-dicarboxylic acid group having from 1 to 7 methylene groups, preferably two methylene groups which form a bridge between an amino group of the parent peptide and an amino group of the lipophilic substituent.
- 8. A GLP-2 derivative of claim 6, wherein the spacer is an amino acid residue except Cys, or a dipeptide such as Gly—Lys.
- 9. A GLP-2 derivative of claim 8, wherein a carboxyl group of the parent peptide forms an amide bond with an amino group of Lys or a dipeptide containing a Lys residue, and the other amino group of the Lys or a dipeptide containing a Lys residue forms an amide bond with a carboxyl group of the lipophilic substituent.
- 10. A GLP-2 derivative of claim 8, wherein an amino group of the parent peptide forms an amide bond with a carboxylic group of the amino acid or dipeptide spacer, and an amino group of the amino acid or dipeptide spacer forms an amide bond with a carboxyl group of the lipophilic substituent.
- 11. A GLP-2 derivative of claim 8, wherein a carboxyl group of the parent peptide forms an amide bond with an amino group of the amino acid or dipeptide spacer, and the carboxyl group of the amino acid or dipeptide spacer forms an amide bond with an amino group of the lipophilic substituent.
- 12. A GLP-2 derivative of claim 8, wherein a carboxyl group of the parent peptide forms an amide bond with an amino group of Asp or Glu, or a dipeptide containing an Asp or Glu residue, and a carboxyl group of the spacer forms an amide bond with an amino group of the lipophilic substituent.
- 13. A GLP-2 derivative of any of the preceding claims, wherein the lipophilic substituent comprises a partially or completely hydrogenated cyclopentanophenathrene skeleton.
- 14. A GLP-2 derivative of any of claims 1-12, wherein the lipophilic substituent is an straight-chain or branched alkyl group.
- 15. A GLP-2 derivative of any of claims 1-12, wherein the lipophilic substituent is the acyl group of a straight-chain or branched fatty acid.
- 16. A GLP-2 derivative of claim 15 wherein the acyl group is selected from the group comprising CH3(CH2),CO—, wherein n is 4 to 38, preferably CH3(CH2)6CO—, CH3(CH2)8CO—, CH3(CH2)10CO—, CH3(CH2)12CO—, CH3(CH2)14CO—, CH3(CH2),6CO—, CH3(CH2)18CO—, CH3(CH2)20CO— and CH3(CH2)22CO—.
- 17. A GLP-2 derivative of any of claims 1-12, wherein the lipophilic substituent is an acyl group of a straight-chain or branched alkane α,ω-dicarboxylic acid.
- 18. A GLP-2 derivative of claim 17 wherein the acyl group is selected from the group comprising HOOC(CH2)mCO—, wherein m is 4 to 38, preferably HOOC(CH2)14CO—, HOOC(CH2)16CO—, HOOC(CH2),8CO—, HOOC(CH20CO— and HOOC(CH2) CO—.
- 19. A GLP-2 derivative of any of claims 1-12, wherein the lipophilic substituent is a group of the formula CH3(CH2)p((CH2)qCOOH)CHNHCO(CH2)2CO wherein p and q are integers and p+q is an integer of from 8 to 40, preferably from 12 to 35.
- 20. A GLP-2 derivative of any of claims 1-12, wherein the lipophlic substitnent is a group of the formula CH3(CH2),CONHCH(COOH)(CH2)2CO, wherein r is an integer of from 10 to 24.
- 21. A GLP-2 derivative of any of claims 1-12, wherein the lipophilic substituent is a group of the formula CH3(CH2),CO—NHCH((CH2)2COOH)CO—, wherein s is an integer of from 8 to 24.
- 22. A GLP-2 derivative of any of claims 1-12, wherein the lipophilic substituent is a group of the formula COOH(CH2),CO— wherein t is an integer of from 8 to 24.
- 23. A GLP-2 derivative of any of claims 1-12, wherein the lipophilic substituent is a group of the formula —NHCH(COOH)(CH2)4NHl—CO(CH2)uCH3, wherein u is an integer of from 8 to 18.
- 24. A GLP-2 derivative of any of claims 1-12, wherein the lipophilic substituent is a group of the formula —NHCH(COOH)(CH2)4NH—COCH((CH2)2COOH)NH—CO(CH2),CH3, wherein w is an integer of from 10 to 16.
- 25. A GLP-2 derivative of any of claims 1-12, wherein the lipophilic substituent is a group of the formula —NHCH(COOH)(CH2)4NH—CO(CH2)2CH(COOH)NH—CO(CH2),CH3, wherein x is an integer of from 10 to 16.
- 26. A GLP-2 derivative of any of claims 1-12, wherein the lipophilic substituent is a group of the formula —NHCH(COOH)(CH2)4NH—CO(CH2)2CH(COOH)NHCO(CH2)yCH3, wherein y is zero or an integer of from 1 to 22.
- 27. A GLP-2 derivative of any of the preceding claims which has one lipophilic substituent.
- 28. A GLP-2 derivative of any of claims 1-26 which has two lipophilic substituents.
- 29. A GLP-2 derivative according any of the preceding claims, wherein the parent peptide is selected from the group comprising GLP-2(1-30); GLP-2(1-31); GLP-2(1-32); GLP-2(1-33); GLP-2(1-34) and GLP-2(1-35) or an analogue or a fragment thereof.
- 30. A GLP-2 derivative of claim 29, wherein the parent peptide is selected from the group comprising GLP-2(1-35) or an analogue or a fragment thereof.
- 31. A GLP-2 derivative of claim 29 or 30 wherein the designation analogue comprises derivatives wherein a total of up to ten amino acid residues have been exchanged with any α-amino acid residue.
- 32. A GLP-2 derivative of any of the preceding claims wherein the parent peptide is selected from the group comprising Lys20GLP-2(1-33); Lys20Arg30GLP-2(1-33); Arg30Lys35GLP-2(1-35); Arg30,35Lys20GLP-2(1-35); Arg35GLP-2(1-35); Arg30Lys34GLP-2(1-34).
- 33. A GLP-2 derivative of claim 1 selected from the group consisting of
Lys2(Nε-tetradecanoyl)GLP-2(1-33); Lys20,30-bis(Nε-tetradecanoyl)GLP-2(1-33); Lys20(Nε-tetradecanoyl)Arg30GLP-2(1-33); Lys20(Nε-tetradecanoyl)Arg30GLP-2(1-33); Arg30Lys35(Nε-tetradecanoyl)GLP-2(1-35); Arg30,35Lys20(Nε-tetradecanoyl)GLP-2(1-35); Arg35Lys30(Nε-tetradecanoyl)GLP-2(1-35); Arg30Lys34(Nε-tetradecanoyl)GLP-2(1-34); Lys20(Nε-(ω-carboxynonadecanoyl))GLP-2(1-33); Lys20,30-bis(Nε-(ω-carboxynonadecanoyl))GLP-2(1-33); Lys20(Nε-(ω-carboxynonadecanoyl))Arg30GLP-2(1-33); Arg30Lys20(Nε-(ω-carboxynonadecanoyl))GLP-2(1-35); Arg35Lys35(Nε-(ω-carboxynonadecanoyl))GLP-2(1-35); Arg35Lys30(Nε-(ω-carboxynonadecanoyl))GLP-2(1-35); and Arg30Lys34(Nε-(ω-carboxynonadecanoyl))GLP-2(1-34).
- 34. A pharmaceutical composition comprising a GLP-2 derivative of any of the preceding claims and a pharmaceutically acceptable vehicle or carrier.
- 35. A method of treating obesity in a patient in need of such a treatment, comprising administering to the patient a therapeutically effective amount of a GLP-2 derivative of any of claims 1-33 together with a pharmaceutically acceptable carrier.
- 36. A method of treating small bowel syndrome in a patient in need of such a treatment, comprising administering to the patient a therapeutically effective amount of a GLP-2 derivative of any of claims 1-33 together with a pharmaceutically acceptable carrier.
- 37. A pharmaceutical composition comprising a GLP-2 derivative which has a helix content as measured by CD at 222 nm in H2O at 22±2° C. exceeding 25%, preferably in the range of 25% to 50%, at a peptide concentration of about 10 μM
- 38. A pharmaceutical composition of claim 37, wherein the concentration of GLP-2 derivative is not less than 0.5 mg/ml, preferably not less than about 5 mg/ml, more preferred not less than about 10 mg/ml and, preferably, not more than about 100 mg/ml.
- 39. A pharmaceutical composition of claim 37 or 38, comprising a GLP-2 derivative wherein at least one amino acid residue of the parent peptide has a lipophilic substituent attached.
- 40. A pharmaceutical composition of claim 39, comprising a GLP-2 derivative having a lipophilic substituent which is attached to any one of the amino acid residues in position 20-34, preferably 30-34, most preferably 30.
- 41. A pharmaceutical composition of any of claims 37-40, further comprising a pharmaceutically acceptable vehicle or carrier.
- 42. A pharmaceutical composition of any of claims 37-41, further comprising an isotonic agent, preferably selected from the group consisting of sodium chloride, mannitol and glycerol.
- 43. A pharmaceutical composition of any of claims 37-42, further comprising a preservative, preferably selected from the group consisting of phenol, m-cresol, methyl p-hydroxybenzoate, butyl p-hydroxybenzoate and benzyl alcohol.
- 44. A pharmaceutical composition of any of claims 37-43, further comprising a buffer, preferably selected from the group consisting of sodium acetate, citrate, glycylglycine, histidine, 2-phenylethanol and sodium phosphate.
- 45. A pharmaceutical composition of any of claims 37-44, further comprising a surfactant capable of improving the solubility and/or the stability of the GLP-2 derivative, preferable selected from poloxymer 188, tween 20 and tween 80.
- 46. A pharmaceutical composition of any of claims 37-45, wherein the parent peptide is selected from the group comprising GLP-2(1-30); GLP-2(1-31); GLP-2(1-32); GLP-2(1-33); GLP-2(1-34) and GLP-2(1-35).
- 47. A pharmaceutical composition of any of claims 37-46, wherein the parent peptide has the following amino acid sequence (SEQ ID NO:1)
X1H X2D G S F S D E M N T X3L D X4L A X X6D F I N W L X7X8T K I T D X9wherein X1 is NH2, DFPEEVAIVEELGRR (SEQ ID NO:2), DFPEEVTIVEELGRR (SEQ ID NO:3), DFPEEVNIVEELRRR (SEQ ID NO:4), or a fragment thereof, X2 is Ala or Gly, X3 is Ile or Val, X4 is Asn, Ser or His, X5 is Ala or Thr, X6 is Arg or Lys, X7 is le or Leu, X8 is Gln or His, and X9 is OH, Lys, Arg, Arg—Lys, Lys—Arg, Arg—Arg or Lys—Lys.
- 48. A pharmaceutical composition of any of claims 37-47, comprising a GLP-2 derivative wherein a total of up to fifteen, preferably up to ten, more preferably up to six, amino acid residues have been exchanged with any a-amino acid residue which can be coded for by the genetic code.
- 49. A pharmaceutical composition of any of claims 3748, wherein the parent peptide is selected from the group comprising Lys20GLP-2(1-33); Lys20Arg30GLP-2(1-33); Arg30Lys34GLP-2(1-34); Arg30Lys35GLP-2(1-35); Arg30,35Lys20GLP-2(1-35); Arg30GLP-2(1-35).
- 50. A method for improving the solubility and/or stability of GLP-2 or a fragment or an analogue thereof, comprising introducing a lipophilic substituent on any one of the amino acid residues of the parent peptide.
- 51. A method of claim 50, wherein a lipophilic substituent is introduced on any one of the amino acid residues in position 20-34, preferably 30-34, most preferred 30.
- 52. A method of claim 50 or 51, wherein the lipophilic substituent comprises from 4 to 40 carbon atoms, preferably from 8 to 25 carbon atoms.
- 53. A method of any of claims 50 to 52, wherein the lipophilic substituent is the acyl group of a straight-chain or branched fatty acid.
- 54. A method of claim 53, wherein the acyl group is selected from the group comprising CH3(CH2)nCO—, wherein n is 4 to 38, preferably CH3(CH2)6CO—, CH3(CH2)8CO—, CH3(CH2)10CO—, CH3(CH2),2CO—, CH3(CH2)14CO—, CH3(CH2),6CO—, CH3(CH2),8CO—, CH3(CH2)20CO— and CH3(CH2)22CO—.
- 55. A method of any of claims 50 to 54, wherein the parent peptide is selected from the group comprising Lys20GLP-2(1-33); Lys20Arg30GLP-2(1-33); Arg3Lys34GLP-2(1-34); Arg30Lys35GLP-2(1-35); Arg30,35Lys20GLP-2(1-35); Arg35GLP-2(1-35).
- 56. A method for treating obesity, comprising administering to a subject in need thereof a pharmaceutical composition of any of claims 37 to 49.
- 57. A method for treating small bowel syndrome, Crohn's disease, ileitis, intestinal inflammation, gastric and duodenal ulceration, inflammatory bowel disease (IBD) and intestinal cancer damage therapy, comprising administering to a subject in need thereof a pharmaceutical composition of any of claims 37 to 49.
Priority Claims (3)
Number |
Date |
Country |
Kind |
0931/96 |
Aug 1996 |
DK |
|
1259/96 |
Nov 1996 |
DK |
|
0271/98 |
Feb 1998 |
DK |
|
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application is a continuation-in-part of Ser. No. 08/922,200 filed Sep. 2, 1997 and claims priority of Danish application serial nos. 0931/96, 1259/96 and 0271/98 filed Aug. 30, 1996, Nov. 8, 1996 and Feb. 27, 1998, respectively, and of U.S. provisional application Ser. Nos. 60/035,905, 60/036,226 and 60/085,789 filed Jan. 24, 1997, Jan. 24, 1997 and May 18, 1998, respectively, the contents of which are fully incorporated herein by reference.
Provisional Applications (3)
|
Number |
Date |
Country |
|
60035905 |
Jan 1997 |
US |
|
60036226 |
Jan 1997 |
US |
|
60085789 |
May 1998 |
US |
Continuations (1)
|
Number |
Date |
Country |
Parent |
09258187 |
Feb 1999 |
US |
Child |
09908534 |
Jul 2001 |
US |
Continuation in Parts (1)
|
Number |
Date |
Country |
Parent |
08922200 |
Sep 1997 |
US |
Child |
09258187 |
Feb 1999 |
US |