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

• US 9,453,064 B2
• Filed: 05/03/2013
• Issued: 09/27/2016
• Est. Priority Date: 05/03/2012
• Status: Active Grant

• Current Assignee: Zealand Pharma A/S
• Original Assignee: Zealand Pharma A/S
• Inventors: Bovbjerg, Kirsten Lindegaard, Russell, Wayne Shaun, Just, Rasmus, Riber, Ditte
• Primary Examiner(s): LIEB, JEANETTE M
• Application Number:
  US14/398,297
• Publication Number:
  US 20150125431A1
• Time in Patent Office: 1,243 Days
• Field of Search: None
• US Class Current: 1/1
• CPC Class Codes: A61K 35/22 Urine; Urinary tract, e.g. kidney or bladder; Intraglomerular mesangial cells; Renal mesenchymal cells; Adrenal gland
• CPC Class Codes: A61K 35/48 Reproductive organs
• CPC Class Codes: A61K 35/54 Ovaries; Ova; Ovules; Embryos; Foetal cells; Germ cells
• CPC Class Codes: A61K 35/63 Arthropods aquatic crustaceans A61K35/612
• CPC Class Codes: A61K 35/64 Insects, e.g. bees, wasps or fleas
• CPC Class Codes: A61K 38/26 Glucagons
• CPC Class Codes: A61K 45/06 Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
• 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 19/02 for joint disorders, e.g. arthritis, arthrosis
• CPC Class Codes: A61P 25/24 Antidepressants
• CPC Class Codes: A61P 25/28 for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
• 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 3/06 Antihyperlipidemics
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 9/00 Drugs for disorders of the cardiovascular system
• CPC Class Codes: A61P 9/10 : for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
• CPC Class Codes: A61P 9/12 : Antihypertensives
• CPC Class Codes: C07K 14/605 : Glucagons

Abstract

GLP-2 analogs are disclosed which comprise one of more substitutions as compared to h[Gly2]GLP-2 and which may have the property of an altered GLP-1 activity, and their medical use. The analogs are particularly useful for the prophylaxis, treatment or ameliorating of the gastro-intestinal associated side effects of diabetes.

Claims

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

R¹-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-R² 

 

(I)

or a pharmaceutically acceptable salt thereof, wherein;

R¹ 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

R² is NH₂ 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

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

R¹ 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

R² is NH₂ 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

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.

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

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 R¹ 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.

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

R¹-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-R² 

 

(1a)

or a pharmaceutically acceptable salt thereof, wherein;

R¹ 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

R² is NH₂ 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

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

R¹ 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

R² is NH₂ 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

36. The analogue of claim 3, wherein R¹ is methyl.

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

R¹-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-R² 

 

(II)

or a pharmaceutically acceptable salt thereof, wherein;

R¹ 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

R² is NH₂ 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:

R¹ 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

R² is NH₂ 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.

37. The analogue of claim 14, wherein R¹ is methyl.

38. The analogue of claim 16, wherein R¹ is methyl.

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

R¹-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-R² 

 

(III)

or a pharmaceutically acceptable salt thereof, wherein;

R¹ 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

R² is NH₂ 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.

39. The analogue of claim 19, wherein R¹ is methyl.