GLUCAGON-LIKE-PEPTIDE-2 (GLP-2) ANALOGUES

• US 20150368314A1
• Filed: 09/08/2015
• Published: 12/24/2015
• Est. Priority Date: 05/04/2005
• Status: Active Grant

• Current Assignee: Zealand Pharma A/S
• Original Assignee: Zealand Pharma A/S
• Inventors: LARSEN, Bjarne Due, Petersen, Yvette Miata
• Granted Patent: US 9,580,487 B2
• Time in Patent Office: Days
• US Class Current: 1/1
• CPC Class Codes: A61K 35/54 Ovaries; Ova; Ovules; Embryos; Foetal cells; Germ cells
• CPC Class Codes: A61K 35/74 Bacteria therapeutic use of a bacterial protein A61K38/00
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: A61K 38/26 Glucagons
• CPC Class Codes: A61K 47/26 Carbohydrates, e.g. sugar alcohols, amino sugars, nucleic acids, mono-, di- or oligo-saccharides; Derivatives thereof, e.g. polysorbates, sorbitan fatty acid esters or glycyrrhizin
• CPC Class Codes: A61K 48/00 Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
• CPC Class Codes: A61K 9/0019 Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner non-active ingredients are additionally classified in A61K47/00
• CPC Class Codes: A61K 9/19 lyophilised , i.e. freeze-dried, solutions or dispersions lyophilised products with subsequent particle size reduction A61K9/14; granules or pellets made by lyphilisation A61K9/1682; solid oral dosage forms made by lyophilisation A61K9/2095; lyophilisation additives A61K47/00
• CPC Class Codes: A61P 1/00 Drugs for disorders of the alimentary tract or the digestive system
• CPC Class Codes: A61P 1/04 for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
• CPC Class Codes: A61P 1/12 Antidiarrhoeals
• CPC Class Codes: A61P 1/14 Prodigestives, e.g. acids, enzymes, appetite stimulants, antidyspeptics, tonics, antiflatulents
• CPC Class Codes: A61P 19/10 for osteoporosis
• CPC Class Codes: A61P 29/00 Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• CPC Class Codes: A61P 3/00 Drugs for disorders of the metabolism of the blood or the extracellular fluid A61P7/00
• CPC Class Codes: A61P 3/04 Anorexiants; Antiobesity agents
• CPC Class Codes: A61P 31/04 Antibacterial agents
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 39/00 General protective or antinoxious agents
• CPC Class Codes: A61P 39/02 Antidotes
• CPC Class Codes: A61P 43/00 : Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 5/00 : Drugs for disorders of the endocrine system
• CPC Class Codes: C07K 14/605 : Glucagons
• CPC Class Codes: C12N 2710/10043 : viral genome or elements thereof as genetic vector
• CPC Class Codes: C12N 2710/20043 : viral genome or elements thereof as genetic vector
• CPC Class Codes: C12N 2710/22043 : viral genome or elements thereof as genetic vector
• CPC Class Codes: C12N 2760/20243 : viral genome or elements thereof as genetic vector
• CPC Class Codes: C12N 7/00 : Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof preparing medicinal viral antigen or antibody compositions, e.g. virus vaccines, A61K39/00

Abstract

GLP-2 analogues are disclosed which comprise one of more substitutions as compared to [hGly2]GLP-2 and which improved biological activity in vivo and/or improved chemical stability, e.g., as assessed in in vitro stability assays. More particularly, preferred GLP-2 analogues disclosed herein comprise substitutions at one or more of positions 8, 16, 24 and/or 28 of the wild-type GLP-2 sequence, optionally in combination with further substitutions at position 2 (as mentioned in the introduction) and one or more of positions 3, 5, 7, 10 and 11, and/or a deletion of one or more of amino acids 31 to 33 and/or the addition of a N-terminal or C-terminal stabilizing peptide sequence. The analogues are particularly useful for the prophylaxis or treatment of stomach and bowel-related disorders and for ameliorating side effects of chemotherapy. Also disclosed are methods and kits for selecting a patient from populations suited for treatment with GLP-2 analogues.

Claims

1. A glucagon-like peptide 2 (GLP-2) analogue represented by general Formula I:

2. The GLP-2 analogue of claim 1 which is represented by general Formula II:

3. The GLP-2 analogue of claim 1 or claim 2 which is represented by general Formula III:

4. The GLP-2 analogue of any one of claims 1 to 3, wherein the GLP-2 analogue has at least 60% amino acid sequence identity to wild-type GLP-2 (1-33) and has the biological activity of causing an increase in intestinal mass in vivo.

5. The GLP-2 analogue of any one of the preceding claims, wherein the GLP-2 analogue comprises more than one of the substitutions at positions X8, X16, X24 and/or X28 and/or one of more of said substitutions in combination with one or more substitutions at positions X3, X5, X7, X10 and/or X11.

6. The GLP-2 analogue of claim 5, wherein said substitutions at position X10 is Leu, Nle, or an oxidatively stable Met-replacement amino acid, such as Met(O) or Met(O)₂.

7. The GLP-2 analogue of claim 5, wherein said substitutions at position X11 is Ala, Ser, or Lys.

8. The GLP-2 analogue of claim 4, wherein the GLP-2 analogue comprises one or more of the following groups of substitutions:

Ser8, Ala16

Ser8, Ala24

Ser8, Ala28

Ala16, Ala24

Ala16, Ala28

Ala24, Ala28

Ser8, Ala16, Ala24

Ser8, Ala16, Ala28

Ser8, Ala24, Ala28

Ala16, Ala24, Ala28

Ser8, Ala16, Ala24, Ala28

9. The GLP-2 analogue of claim 5, wherein the GLP-2 analogue comprises one or more of the following groups of substitutions:

Glu3, Leu10, Ala11,24

Glu3, Thr5, Leu10, Ser11, Ala16,24,28

Glu3, Thr5, Leu10, Lys11, Ala16,24,28

Glu3, Thr5, Ser8, Leu10, Lys11, Ala16,24,28

Glu3, Thr5, Ser8,11, Leu10, Ala16,24,28

Glu3, Thr5, Ser8,11, Leu10, Ala16,24,28

Glu3, Ser8,11, Leu10, Ala16,24,28

Glu3, Leu10, Ser11, Ala16,24,28

Glu3, Leu10, Lys11, Ala16,24,28

Glu3, Thr5, Leu10, Ala11,16,24,28

Glu3, Thr5, Leu10, Ala11,16,24,28, Ile21

Glu3, Thr5, Ser8, Leu10, Ala11,16,24,28

Glu3, Ser8, Leu10, Ala11,16,24,28

Glu3, Leu10, Ala11,16,24,28

Thr7, Leu10, Ala11,24

Thr7, Leu10, Lys11, Ala24

Thr7, Leu10, Ser11, Ala24

Thr7, Leu10, Ser8,11, Ala24

Thr7, Ser8, Leu10, Ala11,24

Thr7, Ser8, Leu10, Lys11, Ala24

Ser8, Leu10, Ala11,24

Leu10, Ala24

Leu10, Ala11, Ala24

Leu10, Ala11,24,28

Leu10, Ala11,16,24,28

Leu10, Lys11, Ala24

Leu10, Ser11, Ala24

Leu10, Ser8,11, Ala24;

or

a deletion at one or more of positions X31-X33.

10. The GLP-2 analogue of any of the preceding claims which is disclosed in Table 1 herein or a pharmaceutically acceptable salt or derivative thereof.

11. The GLP-2 analogue of claim 10 which is:

12. The GLP-2 analogue of any one of claims 1 to 9, wherein the GLP-2 analogue comprises more than one of the substitutions at positions X3, X7, X16, X24, X28, X31, X32 and/or X33.

13. The GLP-2 analogue of claim 12, wherein the GLP-2 analogue comprises one or more of substitutions selected from X3 is Glu, X7 is Ser, X16 is Ala, X24 is Ala, X28 is Ala, X31 is Ile, X32 is Thr and X33 is Asp, and the amino acid residues in positions X31, X32 and X33 are optionally deleted;

or a pharmaceutically acceptable salt or derivative thereof.

14. The GLP-2 analogue of claim 12 or claim 13 which is:

1827 H-HGDGSFTDELSTILDNLAARDFIAWLIQTKKKKKKK-NH2

1844 H-HGEGTFSSELSTILDALAARDFIAWLIATKITDKKKKKK-NH2

1845 H-HGEGTFSDELSTILDALAARDFIAWLIATKITDKKKKKK-NH2

1846 H-HGEGSFSSELSTILDALAARDFIAWLIATKITDKKKKKK-NH2

1848 H-HGEGTFSSELATILDALAARDFIAWLIATKITDKKKKKK-NH2

1849 H-HGEGSFSSELATILDALAARDFIAWLIATKITDKKKKKK-NH2

1850 H-HGEGSFSDELKTILDALAARDFIAWLIATKITDKKKKKK-NH2

1851 H-HGEGSFSDELKTILDALAARDFIAWLIATKITDKKKKKK-NH2

1852 H-HGEGTFSSELKTILDALAARDFIAWLIATKITDKKKKKK-NH2

1855 H-HGEGSFSSELSTILDALAARDFIAWLIATKITD-NH2

1857 H-HGEGTFSSELATILDALAARDFIAWLIATKITD-NH2

1858 H-HGEGSFSSELATILDALAARDFIAWLIATKITD-NH2

1859 H-HGEGSFSDELKTILDALAARDFIAWLIATKITD-NH2;

or a pharmaceutically acceptable salt or derivative thereof.

15. The GLP-2 analogue of any one of claims 1 to 9, wherein the GLP-2 analogue comprises more than one of the substitutions at positions X3, X8 and/or X24.

16. The GLP-2 analogue of claim 15, wherein the GLP-2 analogue comprises one or more of substitutions selected from X3 is Asp, X8 is Asp and X24 is Ala;

and the amino acid residues in positions X31, X32 and X33 are optionally deleted;

or a pharmaceutically acceptable salt or derivative thereof.

17. The GLP-2 analogue of claim 15 or claim 16 which is:

1830 H-HGDGSFSDELSTILDNLAARDFIAWLIQTK-NH2

1831 H-HGDGSFTDELSTILDNLAARDFIAWLIQTK-NH2

1835 H-HGDGSFSDELKTILDNLAARDFIAWLIQTK-NH2

1836 H-HGDGSFTDELKTILDNLAARDFIAWLIQTK-NH2

1839 H-HGDGSFSDELATILDNLAARDFIAWLIQTKITDKKKKKK-NH2

1840 H-HGDGSFSDELATILDNLAARDFIAWLIQTKITD-NH2

1841 H-HGDGSFSDELATILDNLAARDFIAWLIQTK-NH2

1843 H-HGDGSFTDELATILDNLAARDFIAWLIQTK-NH2;

or a pharmaceutically acceptable salt or derivative thereof.

18. The GLP-2 analogue of any one of claims 1 to 9 which possesses a substitution at one or more of positions X3, X33, X10, X11, X16 and/or X24.

19. A GLP-2 analogue of any one of the preceding claims for use in therapy.

20. A pharmaceutical composition comprising a GLP-2 analogue of any one of the preceding claims, or a salt or derivative thereof, in admixture with a carrier.

21. The pharmaceutical composition of claim 20, wherein the GLP-2 analogue is a pharmaceutically acceptable acid addition salt.

22. The pharmaceutical composition of claim 20 or claim 21, which is formulated as a liquid suitable for administration by injection or infusion, or which is formulated to cause slow release of said GLP-2 analogue.

23. Use of a GLP-2 analogue of any one of claims 1 to 18 for the preparation of a medicament for the treatment and/or prevention of a stomach and bowel-related disorder.

24. The use of claim 23, wherein the stomach and bowel-related disorder is ulcers, digestion disorders, malabsorption syndromes, short-gut syndrome, cul-de-sac syndrome, inflammatory bowel disease, celiac sprue (for example arising from gluten induced enteropathy or celiac disease), tropical sprue, hypogammaglobulinemic sprue, enteritis, regional enteritis (Crohn'"'"'s disease), ulcerative colitis, small intestine damage or short bowel syndrome.

25. The use of claim 23, wherein the stomach and bowel-related disorder is radiation enteritis, infectious or post-infectious enteritis, or small intestinal damage due to toxic or other chemotherapeutic agents.

26. Use of a GLP-2 analogue of any one of claims 1 to 18 for the preparation of a medicament for the treatment and/or prevention of a side effect of chemotherapy or radiation treatment.

27. The use of claim 26, wherein the side effect of chemotherapy is diarrhea, abdominal cramping, vomiting or structural and functional damage of the intestinal epithelium resulting from chemotherapy treatment.

28. Use of a GLP-2 analogue of any one of claims 1 to 18 for the preparation of a medicament for the treatment of neo-natals, osteoporosis or DPP-IV (dipeptidylpeptidase-IV) mediated conditions.

29. Use of a GLP-2 analogue of any one of claims 1 to 18 for the preparation of a medicament for the treatment and/or prevention of a condition involving malnutrition.

30. The use of claim 29, wherein the condition involving malnutrition is cachexia or anorexia.

31. A nucleic acid molecule comprising a nucleic acid sequence encoding a GLP-2 analogue of any one of claims 1 to 18.

32. An expression vector comprising the nucleic acid sequence of claim 31, in combination with control sequences to directed its expression.

33. A host cell transformed the expression vector of claim 32.

34. A method of producing the GLP-2 analogue of any one of claims 1 to 18, the method comprising culturing the host cells of claim 22 under conditions suitable for expressing the GLP-2 analogue and purifying the GLP-2 analogue thus produced.

35. A nucleic acid molecule according to claim 31, an expression vector according to claim 32, or a host cell according to claim 33, for use in therapy.

36. Use of a nucleic acid molecule according to claim 31, an expression vector according to claim 32, or a host cell according to claim 33, in the preparation of a medicament for the treatment and/or prevention of a stomach and bowel-related disorder, or for the treatment and/or prevention of a side effect of chemotherapy or radiation treatment, or for the treatment of neo-natals, osteoporosis or DPP-IV (dipeptidylpeptidase-IV) mediated conditions.

37. A method of treating a stomach and bowel-related disorder in a patient in need thereof by administering an effective amount a GLP-2 analogue of any one of claims 1 to 17, a nucleic acid molecule according to claim 31, an expression vector according to claim 32, or a host cell according to claim 33.

38. The method of claim 37, wherein the stomach and bowel-related disorder is ulcers, digestion disorders, malabsorption syndromes, short-gut syndrome, cul-de-sac syndrome, inflammatory bowel disease, celiac sprue (for example arising from gluten induced enteropathy or celiac disease), tropical sprue, hypogammaglobulinemic sprue, enteritis, regional enteritis (Crohn'"'"'s disease), ulcerative colitis, small intestine damage or short bowel syndrome.

39. The method of claim 37, wherein the stomach and bowel-related disorder is radiation enteritis, infectious or post-infectious enteritis, or small intestinal damage due to toxic or other chemotherapeutic agents.

40. A method of treating or preventing a side effect of chemotherapy or radiation therapy to a patient in need thereof, the method comprising administering an effective amount a GLP-2 analogue of any one of claims 1 to 18, a nucleic acid molecule according to claim 31, an expression vector according to claim 32, or a host cell according to claim 33.

41. The method of claim 40, wherein the side effect of chemotherapy is diarrhea, abdominal cramping, vomiting or structural or functional damage of the intestinal epithelium resulting from chemotherapy treatment.

42. A method of treating neo-natal disorders, obesity, osteoporosis or DPP-IV (dipeptidylpeptidase-IV) mediated conditions in a patient in need thereof, the method comprising administering an effective amount a GLP-2 analogue of any one of claims 1 to 18, a nucleic acid molecule according to claim 31, an expression vector according to claim 32, or a host cell according to claim 33.

43. A therapeutic kit comprising a cancer chemotherapy drug and a GLP-2 analogue according to any one of claims 1 to 18, a nucleic acid molecule according to claim 31, an expression vector according to claim 32, or a host cell according to claim 33, each optionally in combination with a pharmaceutically acceptable carrier.

44. A pharmaceutical composition comprising a cancer chemotherapy drug and a GLP-2 analogue according to any one of claims 1 to 18, a nucleic acid molecule according to claim 31, an expression vector according to claim 32, or a host cell according to claim 33 in combination with a pharmaceutically acceptable carrier.

45. A GLP-2 analogue of any of claims 1 to 17 wherein said analog has enhanced stability relative to Gly2-GLP-2.

46. A GLP-2 analogue of any of claims 1 to 17 which is further characterized in having an observed purity of at least 70% relative to the initial purity in conditions selected from the group consisting of 0.1 M HCl after 12 days, 0.5% H₂O₂ after 3 days, and 0.1 M NH₄HCO₃ after 6 days

47. A GLP-2 analogue of any of claims 1 to 17 which is further characterized in having an observed purity of at least 60% relative to initial purity in a solution of HCl 0.1 M after 12 days.

48. A GLP-2 analogue of any of claims 1 to 17 which is further characterized in having an observed purity of at least 70% relative to initial purity in a solution of NH₄HCO₃ 0.1 M after 6 days.