ACYLATED GLUCAGON ANALOGUES

• US 20120178670A1
• Filed: 06/24/2010
• Published: 07/12/2012
• Est. Priority Date: 07/13/2009
• Status: Active Grant

• Current Assignee: Zealand Pharma A/S
• Original Assignee: Zealand Pharma A/S
• Inventors: Meier, Eddi, Daugaard, Jens Rosengren, Riber, Ditte, Skovgaard, Marie, Tolborg, Jakob Lind, Kampen, Gita, Bæk, Camilla Ærteberg
• Granted Patent: US 9,156,901 B2
• Time in Patent Office: Days
• US Class Current: 514/1.9
• CPC Class Codes: A61K 38/26 Glucagons
• CPC Class Codes: A61K 47/54 the modifying agent being an organic compound
• CPC Class Codes: A61P 1/16 for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
• 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 3/08 for glucose homeostasis pancreatic hormones A61P5/48
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 5/00 Drugs for disorders of the endocrine system
• CPC Class Codes: A61P 5/48 of the pancreatic hormones
• CPC Class Codes: A61P 7/12 Antidiuretics, e.g. drugs for diabetes insipidus ADH A61P5/10
• 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

The invention provides materials and methods for promoting weight loss or preventing weight gain, and in the treatment of diabetes and associated metabolic disorders. In particular, the invention provides novel acylated glucagon analogue peptides effective in such methods. The peptides may mediate their effect by having increased selectivity for the GLP-1 receptor as compared to human glucagon.

Claims

1. A compound having the formula:

R¹—

Z—

R²

wherein R¹ is H, C_1-4 alkyl, acetyl, formyl, benzoyl or trifluoroacetyl;

R² is OH or NH₂;

and Z is a peptide having the formula I

2. A compound according to claim 1 wherein one or more of said residues X is independently selected from Lys, Glu and Cys.

3. A compound according to claim 1, wherein:

X16 is selected from Glu, Lys and Ser;

X17 is selected from Lys and Cys;

X20 is selected from His, Lys, Arg and Cys;

X24 is selected from Lys, Glu and Ala;

X27 is selected from Leu and Lys; and

/or

X28 is selected from Ser, Arg and Lys.

4. A compound of claim 1, wherein the peptide of formula I includes one or more of the following combinations of residues:

X2 is Aib and X17 is Lys;

X2 is Aib and X17 is Cys;

X2 is Aib and X20 is Cys;

X2 is Aib and X28 is Lys;

X12 is Arg and X17 is Lys;

X12 is Leu and X17 is Lys;

X12 is Lys and X20 is Lys;

X12 is Lys and X17 is Lys;

X16 is Lys and X17 is Lys;

X16 is Ser and X17 is Lys;

X17 is Lys and X20 is Lys;

X17 is Lys and X21 is Asp;

X17 is Lys and X24 is Glu;

X17 is Lys and X27 is Leu;

X17 is Lys and X27 is Lys;

X17 is Lys and X28 is Ser;

X17 is Lys and X28 is Arg;

X20 is Lys and X27 is Leu;

X21 is Asp and X27 is Leu;

X2 is Aib, X12 is Lys and X16 is Ser;

X12 is Lys, X17 is Lys and X16 is Ser;

X12 is Arg, X17 is Lys and X16 is Glu;

X16 is Glu, X17 is Lys and X20 is Lys;

X16 is Ser, X21 is Asp and X24 is Glu;

X17 is Lys, X24 is Glu and X28 is Arg;

X17 is Lys, X24 is Glu and X28 is Lys;

X17 is Lys, X27 is Leu and X28 is Ser;

X17 is Lys, X27 is Leu and X28 is Arg;

X20 is Lys, X24 is Glu and X27 is Leu;

X20 is Lys, X27 is Leu and X28 is Ser;

X20 is Lys, X27 is Leu and X28 is Arg.

X16 is Ser, X20 is His, X24 is Glu and X27 is Leu;

X17 is Lys, X20 is His, X24 is Glu and X28 is Ser;

X17 is Lys, X20 is Lys, X24 is Glu and X27 is Leu;

or

X17 is Cys, X20 is Lys, X24 is Glu and X27 is Leu.

5. A compound of claim 1, wherein the peptide of formula I contains only one amino acid of the type conjugated to the lipophilic substituent.

6. A compound according to claim 5 wherein the peptide contains only one Lys residue, only one Cys residue or only one Glu residue, and wherein the lipophilic substituent is conjugated to that residue.

7. A compound of claim 1, wherein the peptide sequence of formula I comprises one or more intramolecular bridges.

8. A compound according to claim 7 wherein said intramolecular bridge is formed between the side chains of two amino acid residues which are separated by three amino acids in the linear amino acid sequence of formula I.

9. A compound according to claim 8 wherein the intramolecular bridge is formed between the side chains of residue pairs 16 and 20, 17 and 21, 20 and 24, or 24 and 28.

10. A compound of claim 7, wherein the intramolecular bridge is a salt bridge or a lactam ring.

11. A compound of claim 7, wherein the intramolecular bridge involves a pair of residues wherein:

X16 is Glu and X20 is Lys;

X16 is Glu and X20 is Arg;

X16 is Lys and X20 is Glu;

or

X16 is Arg and X20 is Glu;

X17 is Arg and X21 is Glu;

X17 is Lys and X21 is Glu;

X17 is Arg and X21 is Asp;

or

X17 is Lys and X21 is Asp;

X20 is Glu and X24 is Lys;

X20 is Glu and X24 is Arg;

X20 is Lys and X24 is Glu;

or

X20 is Arg and X24 is Glu;

X24 is Glu and X28 is Lys;

X24 is Glu and X28 is Arg;

X24 is Lys and X28 is Glu;

or

X24 is Arg and X28 is Glu.

12. A compound of claim 1, wherein at least one of X16, X17, X20 and X28 is conjugated to a lipophilic substituent.

13. A compound of claim 1, wherein X30 is absent.

14. A compound of claim 1, wherein X30 is present and is conjugated to a lipophilic substituent.

15. A compound of claim 1, wherein the compound has just one lipophilic substituent, at position 16, 17, 20, 24, 27, 28 or 30, preferably at position 16, 17 or 20, particularly at position 17.

16. A compound of claim 1, wherein the compound has precisely two lipophilic substituents, each at one of positions 16, 17, 20, 24, 27, 28 or 30.

17. A compound according to claim 16 wherein the compound has lipophilic substituents at positions 16 and 17, 16 and 20, 16 and 24, 16 and 27, 16 and 28 or 16 and 30;

at 17 and 20, 17 and 24, 17 and 27, 17 and 28 or 17 and 30;

at 20 and 24, 20 and 27, and 28 or 20 and 30;

at 24 and 27, 24 and 28 or 24 and 30;

at 27 and 28 or 27 and 30;

or at 28 and 30.

18. A compound according to claim 1, having the formula:

R¹—

Z—

R²

wherein R¹ is H, C_1-4 alkyl, acetyl, formyl, benzoyl or trifluoroacetyl;

R² is OH or NH₂;

and Z is a peptide having the formula IIa

19. A compound according to claim 1 having the formula:

R¹—

Z—

R²

wherein R¹ is H, C_1-4 alkyl, acetyl, formyl, benzoyl or trifluoroacetyl;

R² is OH or NH₂;

and Z is a peptide having the formula IIb

20. A compound according to claim 18, having the formula:

R¹—

Z—

R²

wherein R¹ is H, C_1-4 alkyl, acetyl, formyl, benzoyl or trifluoroacetyl;

R² is OH or NH₂;

and Z is a peptide having the formula IIIa

His-Aib-Gln-Gly-Thr-Phe-Thr-Ser-Asp-Tyr-Ser-X12-Tyr-Leu-Asp-Ser-X17-Ala-Ala-X20-X21-Phe-Val-X24-Trp-Leu-Leu-X28-Ala;

(SEQ ID NO;

8) (IIIa)

wherein

X12 is selected from Lys And Arg;

X17 is X;

X20 is selected from His and X;

X21 is selected from Asp and Glu;

X24 is selected from Ala and Glu;

X28 is selected from Ser, Lys and Arg;

and wherein each residue X is independently selected from Glu, Lys, and Cys;

wherein the side chain of at least one residue X is conjugated to a lipophilic substituent having the formula;

(i) Z¹, wherein Z¹ is a lipophilic moiety conjugated directly to the side chain of X;

or

(ii) Z¹Z², wherein Z¹ is a lipophilic moiety, Z² is a spacer, and Z¹ is conjugated to the side chain of X via Z².

21. A compound according to claim 19, having the formula:

R¹—

Z—

R²

wherein R¹ is H, C_1-4 alkyl, acetyl, formyl, benzoyl or trifluoroacetyl;

R² is OH or NH₂;

and Z is a peptide having the formula IIIb

22. A compound according to claim 20, having the formula:

R¹—

Z—

R²

wherein R¹ is H, C_1-4 alkyl, acetyl, formyl, benzoyl or trifluoroacetyl;

R² is OH or NH₂;

and Z is a peptide having the formula IVa

23. A compound according to claim 21, having the formula:

R¹—

Z—

R²

wherein R¹ is H, C_1-4 alkyl, acetyl, formyl, benzoyl or trifluoroacetyl;

R² is OH or NH₂;

and Z is a peptide having the formula IVb

24. A compound of claim 1, wherein the peptide of formula I has the sequence:

25. A compound of claim 1, comprising a peptide of formula I having the sequence:

26. A compound of claim 1, wherein Z¹ comprises a hydrocarbon chain having 10 to 24 C atoms, 10 to 22 C atoms, or 10 to 20 C atoms.

27. A compound according to claim 26 wherein Z¹ is a dodecanoyl, 2-butyloctanoyl, tetradecanoyl, hexadecanoyl, heptadecanoyl, octadecanoyl or eicosanoyl moiety.

28. A compound of claim 1, wherein Z² is or comprises one or more amino acid residues.

29. A compound according to claim 28 wherein Z² is a γ

-Glu, Glu, β

-Ala or ε

-Lys residue, or a 3-aminopropanoyl, 4-aminobutanoyl, 8-aminooctanoyl or 8-amino-3,6-dioxaoctanoyl moiety.

30. A compound according to claim 29 wherein the lipophilic substituent is selected from the group consisting of dodecanoyl-γ

-Glu, hexadecanoyl-γ

-Glu, hexadecanoyl-Glu, hexadecanoyl-[3-aminopropanoyl], hexadecanoyl-[8-aminooctanoyl], hexadecanoyl-ε

-Lys, 2-butyloctanoyl-γ

-Glu, octadecanoyl-γ

-Glu and hexadecanoyl-[4-aminobutanoyl].

31. A compound according to claim 30 wherein Z has the formula:

32. A compound according to claim 30 wherein Z has the formula:

37. A pharmaceutically acceptable composition comprising a compound of claim 1, or a salt or derivative thereof, in a mixture with a pharmaceutically acceptable carrier.

40. A method of diminishing weight gain or promoting weight loss in a subject, by administering a compound of claim 1 to said subject, in an amount sufficient to diminish weight gain or promote weight loss.

41. A method of improving circulating glucose levels, glucose tolerance and/or circulating cholesterol levels, lowering circulating LDL levels, and/or increasing HDL/LDL ratio in a subject, by administering a compound of claim 1 to said subject, in an amount sufficient to improve circulating glucose levels, glucose tolerance and/or circulating cholesterol levels, lowering circulating LDL levels, and/or increasing HDL/LDL ratio.

42. A method of treatment of a condition caused or characterized by excess body weight, and the treatment of obesity, morbid obesity, obesity linked inflammation, obesity linked gallbladder disease, obesity induced sleep apnea, metabolic syndrome, pre-diabetes, insulin resistance, glucose intolerance, type 2 diabetes, type I diabetes, hypertension, atherogenic dyslipidaemia, atherosclerosis, arteriosclerosis, coronary heart disease, peripheral artery disease, stroke or microvascular disease in a subject, by administering a compound of claim 1 to said subject, in an amount sufficient to treat a condition caused or characterized by excess body weight, and to treat obesity, morbid obesity, obesity linked inflammation, obesity linked gallbladder disease, obesity induced sleep apnea, metabolic syndrome, pre-diabetes, insulin resistance, glucose intolerance, type 2 diabetes, type I diabetes, hypertension, atherogenic dyslipidaemia, atherosclerosis, arteriosclerosis, coronary heart disease, peripheral artery disease, stroke or microvascular disease.

46. The method of claim 40, wherein the compound is administered as part of a combination therapy with an agent for treatment of diabetes, obesity, dyslipidaemia, or hypertension.

47. The method of claim 46 wherein:

a.) the agent for treatment of diabetes is metformin, a sulfonylurea, a glinide, a DPP-IV inhibitor, a glitazone, insulin or an insulin analogue;

b.) the agent for treatment of obesity is a glucagon-like peptide receptor 1 agonist, peptide YY or analogue thereof, cannabinoid receptor 1 antagonist, lipase inhibitor, melanocortin receptor 4 agonist, or melanin concentrating hormone receptor 1 antagonist;

c.) the agent for treatment of hypertension is an angiotensin-converting enzyme inhibitor, angiotensin II receptor blocker, diuretic, beta-blocker, or calcium channel blocker and;

d.) the agent for treatment of dyslipidaemia is a statin, a fibrate, a niacin, and/or a cholesterol absorption inhibitor.

33. A compound having the formula:

R¹—

Z—

R²

wherein R¹ is H, C_1-4 alkyl, acetyl, formyl, benzoyl or trifluoroacetyl;

R² is OH or NH₂;

and Z is a peptide having the formula V

34. A compound according to claim 33 wherein Z has the formula:

35. A compound having the formula:

R¹—

Z—

R²

wherein R¹ is H, C_1-4 alkyl, acetyl, formyl, benzoyl or trifluoroacetyl;

R² is OH or NH₂;

and Z is a peptide having the formula VI

36. A compound according to claim 35 wherein Z has the formula:

38-39. -39. (canceled)

43-45. -45. (canceled)

48-50. -50. (canceled)