Glucagon-like-peptide-2 (GLP-2) analogues

1. A GLP-2 analogue represented by the general Formula I:

R 1 -His-X2-X3-Gly-X5-Phe-X7-X8-X9-X10-X11-X12-X13-X14-X15-X16-X17-Ala-X19-X20-X21-Phe-Ile-X24-Trp-Leu-X27-X28-X29-X30-X31-X32-X33-X34-R 2   (I)

or a pharmaceutically acceptable salt thereof, wherein: R 1 is hydrogen, C 1-4 alkyl, acetyl, formyl, benzoyl or trifluoroacetyl;

X2 is Gly, Ala or Aib;

X3 is Glu, Gln or Asp;

X5 is Ser or Thr;

X7 is Ser or Thr;

X8 is Asp, Glu or Ser;

X9 is Glu or Asp;

X10 is Met, Val, Leu or Tyr;

X11 is Asn, Ser or Ala;

X12 is Thr, Ser or Lys;

X13 is Ile, Leu, Val, Tyr, Phe or Gln;

X14 is Leu or Met;

X15 is Asp or Glu;

X16 is Asn, Gln, Gly, Ser, Ala, Glu or Lys;

X17 is Gln, Lys, Arg, His or Glu;

X19 is Ala or Val;

X20 is Arg, Lys or His;

X21 is Asp, Glu or Leu;

X24 is Asn, Ala, Glu or Lys;

X27 is Ile, Leu, Val, Glu or Lys;

X28 is Gln, Asn, Lys, Ser, Gly, Y1 or absent;

X29 is Thr, Ala, Y1 or absent;

X30 is Lys, Y1 or absent

X31 is Ile, Pro, Y1 or absent;

X32 is Thr, Y1 or absent;

X33 is Asp, Asn, Y1 or absent;

X34 is Y1 or absent;

Y1 is Gly-Gly-Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro-Ser, or Lys-Asn-Gly-Gly-Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro-Ser; and

R 2 is NH 2 or OH; wherein

the GLP-2 analogue contains no more than one Y1;

if any of X28 to X33 is Y1, those positions X29 to X34 downstream of that Y1 are absent;

if any of X28 to X33 is absent, those positions X29 to X33 downstream of that position are also absent; and with the proviso that the GLP-2 analogue of Formula I is not

HGDGSFSDEMNTILDGQAARDFINWLIQTKITD;

(SEQ ID NO: 3)

HGDGSFSDEMNTILDNQAARDFINWLIQTKITD;

(SEQ ID NO: 4)

or

HGDGSFSDEMNTILDSQAARDFINWLIQTK.

(SEQ ID NO: 5)

2. The GLP-2 analogue or a pharmaceutically acceptable salt thereof according to claim 1 , wherein: R 1 is hydrogen,

X2 is Gly, Ala or Aib;

X3 is Glu, Gln or Asp;

X5 is Ser or Thr;

X7 is Ser or Thr;

X8 is Asp, Glu or Ser;

X9 is Glu or Asp;

X10 is Met, Val, Leu or Tyr;

X11 is Asn or Ser;

X13 is Ile, Leu, Val, Tyr, Phe or Gln;

X14 is Leu or Met;

X12 is Thr or Lys;

X15 is Asp or Glu;

X16 is Asn, Gln, Gly, Ser, Ala, Glu or Lys;

X17 is Gln or Lys;

X19 is Ala or Val;

X20 is Arg, Lys or His;

X21 is Asp, Glu or Leu;

X24 is Asn, Ala, Glu or Lys;

X27 is Ile, Leu, Val or Lys;

X28 is Gln, Asn, Gly, Y1 or absent;

X29 is Thr, Ala, Y1 or absent;

X30 is Lys, Y1 or absent;

X31 is Ile, Pro, Y1 or absent

X32 is Thr, Y1 or absent;

X33 is Asp, Asn, Y1 or absent;

X34 is Y1 or absent;

Y1 is -Gly-Gly-Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro-Ser;

and

R 2 is NH 2 or OH; wherein

the GLP-2 analogue contains no more than one Y1;

if any of X28 to X33 is Y1, those positions X29 to X34 downstream of that Y1 are absent;

if any of X28 to X33 is absent, those positions X29 to X33 downstream of that position are also absent; and with the proviso that the GLP-2 analogue of Formula I is not

HGDGSFSDEMNTILDGQAARDFINWLIQTKITD;

(SEQ ID NO: 3)

HGDGSFSDEMNTILDNQAARDFINWLIQTKITD;

(SEQ ID NO: 4)

or

HGDGSFSDEMNTILDSQAARDFINWLIQTK.

(SEQ ID NO: 5)

3. A GLP-2 analogue represented by the general Formula 1a:

R 1 -His-X2-X3-Gly-X5-Phe-X7-X8-X9-X10-X11-X12-X13Leu X15-X16-X17-Ala-Ala-X20-X21-Phe-Ile-X24-Trp-Leu-X27-X28-X29-X30-X31-X32-X33-R 2   (1a)

or a pharmaceutically acceptable salt thereof, wherein: R 1 is hydrogen, C 1-4 alkyl, acetyl, formyl, benzoyl or trifluoroacetyl;

X2 is Gly, Ala or Aib;

X3 is Glu, Gln or Asp;

X5 is Ser or Thr;

X7 is Ser or Thr;

X8 is Asp, Glu or Ser;

X9 is Glu or Asp;

X10 is Met, Val, Leu or Tyr;

X11 is Asn or Ser;

X12 is Thr, Ser or Lys;

X13 is Ile, Leu, Val, Tyr, Phe or Gln;

X15 is Asp or Glu;

X16 is Asn, Gln, Gly, Ser, Ala, Glu or Lys;

X17 is Gln, Lys, Arg, His or Glu;

X20 is Arg, Lys or His;

X21 is Asp, Glu or Leu

X24 is Asn, Ala, Glu or Lys;

X27 is Ile, Leu, Val, Glu or Lys;

X28 is Gln, Asn, Lys, Ser, Gly, Y1 or absent;

X29 is Thr, Y1 or absent;

X30 is Lys, Y1 or absent;

X31 is Ile, Pro, Y1 or absent;

X32 is Thr, Y1 or absent;

X33 is Asp, Asn, Y1 or absent;

Y1 is Gly-Gly-Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro-Ser;

and

R 2 is NH 2 or OH; wherein

the GLP-2 analogue contains no more than one Y1;

if any of X28 to X33 is Y1, those positions X29 to X34 downstream of that Y1 are absent;

if any of X28 to X33 is absent, those positions X29 to X33 downstream of that position are also absent; and with the proviso that the GLP-2 analogue of Formula 1a is not

HGDGSFSDEMNTILDGQAARDFINWLIQTKITD;

(SEQ ID NO: 3)

HGDGSFSDEMNTILDNQAARDFINWLIQTKITD;

(SEQ ID NO: 4)

or

HGDGSFSDEMNTILDSQAARDFINWLIQTK.

(SEQ ID NO: 5)

4. The GLP-2 analogue or pharmaceutically acceptable salt thereof, according to claim 3 wherein:

R 1 is hydrogen,

X2 is Gly, Ala or Aib;

X3 is Glu, Gln or Asp;

X5 is Ser or Thr;

X7 is Ser or Thr;

X8 is Asp, Glu or Ser;

X9 is Glu or Asp;

X10 is Met, Val, Leu or Tyr;

X11 is Asn or Ser;

X12 is Thr or Lys;

X13 is Ile, Leu, Val, Tyr, Phe or Gln;

X15 is Asp or Glu;

X16 is Asn, Gln, Gly, Ser, Ala, Glu or Lys;

X17 is Gln or Lys

X20 is Arg, Lys or His;

X21 is Asp, Glu or Leu;

X24 is Asn, Ala, Glu or Lys;

X27 is Ile, Leu, Val or Lys;

X28 is Gln, Asn, Gly, Y1 or absent;

X29 is Thr, Y1 or absent;

X30 is Lys, Y1 or absent;

X31 is Ile, Pro, Y1 or absent;

X32 is Thr, Y1 or absent;

X33 is Asp, Asn, Y1 or absent;

Y1 is Gly-Gly-Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro-Ser; and

R 2 is NH 2 or OH; wherein

the GLP-2 analogue contains no more than one Y1;

if any of X28 to X33 is Y1, those positions X29 to X34 downstream of that Y1 are absent;

if any of X28 to X33 is absent, those positions X29 to X33 downstream of that position are also absent; and with the proviso that the GLP-2 analogue of Formula 1a is not

HGDGSFSDEMNTILDGQAARDFINWLIQTKITD;

(SEQ ID NO: 3)

HGDGSFSDEMNTILDNQAARDFINWLIQTKITD;

(SEQ ID NO: 4)

or

HGDGSFSDEMNTILDSQAARDFINWLIQTK.

(SEQ ID NO: 5)

5. The GLP-2 analogue according to claim 1 wherein X17 is Gln.

6. The GLP-2 analogue according to claim 1 wherein X17 is Lys.

7. The GLP-2 analogue according to claim 1 wherein X17 is Glu.

8. The GLP-2 analogue according to claim 1 wherein X14 is Leu.

9. The GLP-2 analogue according to claim 1 wherein X14 is Met.

10. The GLP-2 analogue according to claim 1 wherein:

(i) X16 is Gly and X17 is Gln;

(ii) X16 is Gly and X17 is Lys; or

(iii) X16 is Gly and X17 is Glu.

11. The GLP-2 analogue according to claim 1 wherein X2 is Aib.

12. The GLP-2 analogue according to claim 1 wherein X19 is Ala.

13. The GLP-2 analogue according to claim 1 wherein X19 is Val.

14. A GLP-2 analogue represented by the general Formula II:

R 1 -His-X2-X3-Gly-X5-Phe-X7-Ser-Glu-Leu-Ala-X12-X13-X14-X15-X16-X17-Ala-X19-X20-X21-Phe-Ile-X24-Trp-Leu-X27-X28-X29-X30-X31-X32-X33-X34-R 2   (II)

or a pharmaceutically acceptable salt thereof, wherein:

R 1 is hydrogen, C 1-4 alkyl, acetyl, formyl, benzoyl or trifluoroacetyl;

X2 is Gly, Ala or Aib;

X3 is Glu, Gln or Asp;

X5 is Ser or Thr;

X7 is Ser or Thr;

X12 is Thr, Ser or Lys;

X13 is Ile, Leu, Val, Tyr, Phe or Gln;

X14 is Leu or Met;

X15 is Asp or Glu;

X16 is Gly, Ser, Ala, Glu or Lys;

X17 is Gln or Lys;

X19 is Ala or Val;

X20 is Arg, Lys or His;

X21 is Asp, Glu or Leu;

X24 is Asn, Ala, Glu or Lys;

X27 is Ile, Leu, Val, Glu or Lys;

X28 is Gln, Asn, Lys, Ser, Y1 or absent;

X29 is Thr, Ala, Y1 or absent;

X30 is Lys, Y1 or absent;

X31 is Ile, Pro, Y1 or absent;

X32 is Thr, Y1 or absent;

X33 is Asp, Asn, Y1 or absent;

X34 is Y1 or absent;

Y1 is Gly-Gly-Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro-Ser, or Lys-Asn-Gly-Gly-Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro Ser; and

R 2 is NH 2 or OH; wherein the GLP-2 analogue contains no more than one Y1;

if any of X28 to X33 is Y1, those positions X29 to X34 downstream of that Y1 are absent;

if any of X28 to X33 is absent, those positions X29 to X33 downstream of that position are also absent.

15. A GLP-2 analogue according to claim 14 wherein X16 of Formula II is Gly, Ser or Ala.

16. A GLP-2 analogue according to claim 14 wherein:

R 1 is hydrogen, C 1-4 alkyl, acetyl, formyl, benzoyl or trifluoroacetyl;

X2 is Gly or Aib;

X3 is Glu or Asp;

X5 is Ser or Thr;

X7 is Ser or Thr;

X12 is Thr, Ser or Lys;

X13 is Ile, Leu, Val, Tyr, Phe or Gln;

X14 is Leu or Met;

X15 is Asp or Glu;

X16 is Gly, Ser or Ala;

X17 is Gln or Lys;

X19 is Ala or Val;

X20 is Arg, Lys or His;

X21 is Asp, Glu or Leu;

X24 is Asn, Ala, Glu or Lys;

X27 is Ile, Leu, Val, Glu or Lys;

X28 is Gln, Asn, Lys, Set, Y1 or absent;

X29 is Thr, Ala, Y1 or absent;

X30 is Lys, Y1 or absent;

X31 is Ile, Pro, Y1 or absent;

X32 is Thr, Y1 or absent;

X33 is Asp, Asn, Y1 or absent;

X34 is Y1 or absent;

Y1 is Gly-Gly-Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro-Ser, or Lys-Asn-Gly-Gly-Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro-Ser; and

R 2 is NH 2 or OH; wherein

the GLP-2 analogue contains no more than one Y1;

if any of X28 to X33 is Y1, those positions X29 to X34 downstream of that Y1 are absent;

if any of X28 to X33 is absent, those positions X29 to X33 downstream of that position are also absent.

17. A GLP-2 analogue according to claim 14 wherein

(i) X16 is Gly and X17 is Gln; or

(ii) X16 is Gly and X17 is Lys.

18. A GLP-2 analogue according to claim 14 wherein:

(i) X2 is Gly; or

(ii) X2 is Aib.

19. A GLP-2 analogue represented by the general Formula III:

R 1 -His-Gly-X3-Gly-X5-Phe-X7-Ser-Glu-Leu-Ala-X12-X13-Leu-X15-Gly-X17-Ala-X19-X20-X21-Phe-Ile-X24-Trp-Leu-X27-X28-X29-X30-X31-X32-X33-X34-R 2   (III)

or a pharmaceutically acceptable salt thereof, wherein: R 1 is hydrogen, C 1-4 alkyl, acetyl, formyl, benzoyl or trifluoroacetyl;

X3 is Glu or Asp;

X5 is Ser or Thr;

X7 is Ser or Thr;

X12 is Thr, Ser or Lys;

X13 is Ile, Tyr, or Gln;

X15 is Asp or Glu;

X17 is Gln or Lys;

X19 is Ala or Val;

X20 is Arg, Lys or His;

X21 is Asp, Glu or Leu;

X24 is Asn, Ala, or Glu;

X27 is Ile, Leu, Glu or Lys;

X28 is Gln, Lys, Ser, Gly, Y1 or absent;

X29 is Thr, Ala, Y1 or absent;

X30 is Lys, Y1 or absent;

X31 is Ile, Pro, Y1 or absent;

X32 is Thr, Y1 or absent

X33 is Asp, Asn, Y1 or absent;

X34 is Y1 or absent;

Y1 is Gly-Gly-Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro-Ser, or Lys-Asn-Gly-Gly-Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro-Ser; and

R 2 is NH 2 or OH; wherein

the GLP-2 analogue contains no more than one Y1;

if any of X28 to X33 is Y1, those positions X29 to X34 downstream of that Y1 are absent;

if any of X28 to X33 is absent, those positions X29 to X33 downstream of that position are also absent.

20. The GLP-2 analogue of claim 1 , selected from

(SEQ ID NO: 8)

H-Aib-DGSFSDEMNTILDNQAARDFINWLIQTKITD;

(SEQ ID NO: 9)

HGDGSFSDEMNTILDNKAARDFINWLIQTKITD;

(SEQ ID NO: 10)

HGDGSFSDEMNTILDGQAARDFINWLIQTK;

(SEQ ID NO: 11)

HGDGSFSSEMNTILDSQAARDFINWLIQTKITD;

(SEQ ID NO: 12)

HGEGTFTSDLSKQMEGQAVRDFIEWLIQTKITD;

(SEQ ID NO: 13)

HGEGTFTSDLSKQMESKAARDFIEWLIQTKITD;

(SEQ ID NO: 14)

HGDGSFSSELATILDGKAARDFINWLIQTKITD;

(SEQ ID NO: 15)

HGEGTFTSDLSTILENKAARDFIEWLIQTKITD;

(SEQ ID NO: 16)

HGEGSFSSDLSTILENKAARDFIEWLIQTKITD;

(SEQ ID NO: 17)

H-Aib-DGSFSDELNTILDGKAARDFINWLIQTK;

(SEQ ID NO: 18)

HGDGSFSSELATILDGQAARDFIAWLIQTKITD;

(SEQ ID NO: 19)

HGDGSFSDEMNTILDGQAARDFINWLIQTK;

and

(SEQ ID NO: 20)

HGEGSFSSDLSTILEGKAARDFIEWLIQTKITD;

or a pharmaceutically acceptable salt thereof.

21. The GLP-2 analogue according to claim 1 , wherein a lipophilic substituent is conjugated to one or more of positions 12, 14, 16, 17, 19, 20, 24, 27, 28 and 32.

22. A pharmaceutical composition comprising a GLP-2 analogue of claim 1 , or a salt thereof, in admixture with a carrier.

23. The pharmaceutical composition of claim 22 , wherein the GLP-2 analogue is a pharmaceutically acceptable acid addition salt.

24. The pharmaceutical composition of claim 22 , 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.

25. A method of treating or preventing low grade inflammation, said method comprising administering a GLP-2 analogue of claim 1 .

26. The method of claim 25 , wherein said low grade inflammation is related to diabetes.

27. The method of claim 26 , wherein the low grade inflammation related to diabetes is associated with a condition selected from the group consisting of metabolic syndrome; obesity diabetes; cardiovascular diseases; gastrointestinal inflammation; depression; Alzheimer's disease; arthritis; hypertension; dyslipidaemia; stroke; gastro-intestinal disorders in the upper gastrointestinal tract of the esophagus, the stomach, duodenum, the small intestine, colon and rectum; ulcers of any etiology; digestion disorders; malabsorption syndromes; short-bowel syndrome; cul-de-sac syndrome; inflammatory bowel disease; celiac sprue; tropical sprue; hypogammaglobulinemic sprue; and chemotherapy and/or radiation hemotherapy induced mucositis and diarrhea.

28. A nucleic acid molecule comprising a nucleic acid sequence encoding a GLP-2 analogue of claim 1 .

29. An expression vector comprising the nucleic acid sequence of claim 28 , in combination with control sequences to direct its expression.

30. A host cell transformed by the expression vector of claim 29 .

31. A method of producing the GLP-2 analogue of claim 1 , the method comprising culturing a host cell transformed by an expression vector under conditions suitable for expressing the GLP-2 analogue and purifying the GLP-2 analogue thus produced, wherein said expression vector comprises a nucleic acid sequence encoding a GLP-2 analogue of claim 1 and control sequences to direct its expression.

32. A method of treating a gastrointestinal related disorder in a patient in need thereof, said method comprising administering an effective amount a GLP-2 analogue of claim 1 .

33. The method of claim 32 , wherein the gastro-intestinal related disorder is low grade inflammation associated with a condition selected from the group consisting of metabolic syndrome; obesity diabetes; cardiovascular diseases; gastrointestinal inflammation; depression; Alzheimer's disease; arthritis; hypertension; dyslipidaemia; stroke; gastro-intestinal disorders in the upper gastrointestinal tract of the esophagus, the stomach, duodenum, the small intestine, colon and rectum; ulcers of any etiology; digestion disorders; malabsorption syndromes; short-bowel syndrome; cul-de-sac syndrome; inflammatory bowel disease; celiac sprue; tropical sprue; hypogammaglobulinemic sprue; and chemotherapy and/or radiation hemotherapy induced mucositis and diarrhea.

34. A therapeutic kit comprising a cancer chemotherapy drug and a GLP-2 analogue according to claim 1 , optionally in combination with a pharmaceutically acceptable carrier.

35. The analogue of claim 1 , wherein R 1 is methyl.

36. The analogue of claim 3 , wherein R 1 is methyl.

37. The analogue of claim 14 , wherein R 1 is methyl.

38. The analogue of claim 16 , wherein R 1 is methyl.

39. The analogue of claim 19 , wherein R 1 is methyl.

40. The method of claim 27 , wherein the type of obesity is abdominal obesity.

41. The method of claim 33 , wherein the type of obesity is abdominal obesity.