COMBINATION OF ACYLATED GLUCAGON ANALOGUES WITH INSULIN ANALOGUES

• US 20140011733A1
• Filed: 01/20/2012
• Published: 01/09/2014
• Est. Priority Date: 01/20/2011
• Status: Abandoned Application

• Current Assignee: Zealand Pharma A/S
• Original Assignee: Zealand Pharma A/S
• Inventors: Fosgerau, Keld, Riber, Ditte
• Application Number:
  US13/980,087
• Publication Number:
  US 20140011733A1
• Time in Patent Office: Days
• US Class Current: 514/1.9
• CPC Class Codes: A61K 2300/00 Mixtures or combinations of active ingredients, wherein at least one active ingredient is fully defined in groups A61K31/00 - A61K41/00
• CPC Class Codes: A61K 38/26 Glucagons
• CPC Class Codes: A61K 38/28 Insulins
• 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 5/48 of the pancreatic hormones
• 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: C07K 14/605 Glucagons

Abstract

The invention relates to methods for treating metabolic disorders, including diabetes by using a combination of an acylated glucagon analogue and an insulin analogue. The invention also features a kit that includes an acylated glucagon analogue and an insuline analogue.

Claims

1-4. -4. (canceled)

5. A method for preventing or reducing weight gain;

promoting weight loss;

improving circulating glucose levels, glucose tolerance or circulating cholesterol levels;

lowering circulating LDL levels;

increasing HDL/LDL ratio;

or treating a condition caused or characterized by excess body weight, said method comprising administering to a mammalian subject a combination of compounds comprising;

(a) 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;

His-X2-Gln-Gly-Thr-Phe-Thr-Ser-Asp-Tyr-Ser-X12-Tyr-Leu-Asp-X16-X17-Ala-Ala-X20-X21-Phe-Val-X24-Trp-Leu-X27-X28-Ala-X30 (SEQ ID NO;

6);

 

 

(I)

wherein

X2 is selected from Aib and Ser;

X12 is selected from Lys, Arg, or Leu;

X16 is selected from Arg and X;

X17 is selected from Arg and X;

X20 is selected from Arg, His, and X;

X21 is selected from Asp and Glu;

X24 is selected from Ala and X;

X27 is selected from Leu and X;

X28 is selected from Arg and X;

X30 is X or is absent;

wherein at least one of X16, X17, X20, X24, X27, X28, and X30 is X;

and wherein each residue X is independently selected from the group consisting of Glu, Lys, Ser, Cys, Dbu, Dpr, and Orn;

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; and

(b) an insulin analogue.

6. The method of claim 5, wherein said insulin analogue is selected from the group consisting of insulin glulisine, insulin lispro, Degludec, Actraphane HM, LY2963016, LY2605541, pegylated insulin Lispro, insulin glargine, insulin detemir, insulin isophane, insulin aspart, insulin buccal, hyaluronidase insulin, insulin protamine, NN-1953, IN-105, BIOD-620, and Analog-PH20.

9. The method of claim 5, wherein said condition caused or characterized by excess body weight is selected from the group consisting 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, and microvascular disease.

12. The method of claim 5, wherein said subject has type 1 or type 2 diabetes.

13. The method of claim 5, wherein one or more of said residues X is independently selected from Lys, Glu and Cys.

14. The method of claim 5, 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.

15. The method of claim 5, wherein the peptide of formula I includes one or more of a combination of residues selected from the group consisting of:

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, X2 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; and

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

16. The method of claim 5, wherein the peptide of formula I contains only one amino acid of the type conjugated to the lipophilic substituent.

17. The method of claim 5, wherein the peptide of formula I contains only one Lys residue, only one Cys residue, or only one Glu residue, and wherein the lipophilic substituent is conjugated to that residue.

18. The method of claim 5, wherein the peptide sequence of formula I comprises one or more intramolecular bridges.

19. The method of claim 18, wherein the intramolecular bridge is selected from the group consisting of:

a.) an intramolecular bridge 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,

b.) an intramolecular bridge formed between the side chains of residue pairs 16 and 20, 17 and 21, 20 and 24, or 24 and 28,

c.) a salt bridge or a lactam ring, and

d.) an intramolecular bridge which involves a pair of residues selected from the group consisting of;

X16 is Glu and X20 is Lys;

X16 is Glu and X20 is Arg,

X16 is Lys and X20 is Glu;

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;

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;

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; and

X24 is Arg and X28 is Glu.

23. The method of claim 5, wherein at least one of X16, X17, X20, and X28 is conjugated to a lipophilic substituent.

24. The method of claim 5, wherein X30 is present and conjugated to a lipophilic substituent.

26. The method of claim 5, wherein the compound has lipophilic substituents selected from the group consisting of:

a.) 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

b.) precisely two lipophilic substituents, each at one of positions 16, 17, 20, 24, 27, 28, and 30, and

c.) lipophilic substituents at positions 16 and 17, 16 and 20, 16 and 24, 16 and 27, 16 and 28, 16 and 30, 17 and 20, 17 and 24, 17 and 27, 17 and 28, 17 and 30, 20 and 24, 20 and 27, 20 and 28, 20 and 30, 24 and 27, 24 and 28, 24 and 30, 27 and 28, 27 and 30, or 28 and 30.

29. The method of claim 5, wherein said compound has 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 selected from the group consisting of;

a.) a peptide having the formula IIa;

His-Aib-Gln-Gly-Thr-Phe-Thr-Ser-Asp-Tyr-Ser-X12-Tyr-Leu-Asp-X16-X17-Ala-Ala-X20-X21-Phe-Val-X24-Trp-Leu-Leu-X28-Ala (SEQ ID NO;

7);

 

 

(IIa)

wherein;

X12 is selected from Lys, Arg, and Leu;

X16 is selected from Ser and X;

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 the group consisting of 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,

b.) 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;

 

 

(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), 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, and

c.) a peptide having the formula IVa;

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

 

 

(IVa);

wherein

X12 is selected from Lys and Arg;

X17 is X;

X21 is selected from Asp and Glu;

X24 is selected from Ala and Glu;

X28 is selected from Ser, Lys, and Arg,

wherein X is selected from the group consisting of Glu, Lys, and Cys;

and wherein the side chain of 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

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

35. The method of claim 5, wherein said peptide Z has a sequence selected from the group consisting of:

37. The method of claim 5, wherein Z¹ comprises a moiety selected from the group consisting of a hydrocarbon chain having 10 to 24 C atoms, 10 to 22 C atoms, or 10 to 20 C atoms, a dodecanoyl moiety, 2-butyloctanoyl moiety, tetradecanoyl moiety, hexadecanoyl moiety, heptadecanoyl moiety, octadecanoyl moiety, and an eicosanoyl moiety.

39. The method of claim 5, wherein Z² is or comprises a moiety selected from the group consisting of one or more amino acid residues, a γ

-Glu residue, Glu residue, β

-Ala residue, ε

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

41. The method of claim 39, wherein the lipophilic substituent is selected from the group consisting of dodecanoyl-γ

-Glu, hexadecanoly-γ

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

-Lys, 2-butyloctanoyl-γ

-Glu, octadecanoyl-γ

-Glu, and hexadecanoyl-[4-aminobutanoyl].

42. The method of claim 41, wherein Z has a formula selected from the group consisting of:

71. The method of claim 5, wherein said combination of compounds is selected from the group consisting of:

H—

H-Aib-QGTFTSDYSKYLDS-K(Hexadecanoyl-isoGlu)-AAHDFVEWLLSA-NH₂ (SEQ ID NO;

126) and insulin glargine, H—

H-Aib-QGTFTSDYSKYLDS-K(Hexadecanoyl-isoGlu)-AAHDFVEWLLSA-(SEQ ID NO;

126) and insulin detemir, H—

H-Aib-QGTFTSDYSKYLDS-K(Hexadecanoyl-isoGlu)-AAHDFVEWLLSA-NH, (SEQ ID NO;

126) and glulisine, H—

H-Aib-QGTFTSDYSKYLDS-K(Hexadecanoyl-isoGlu)-AAHDFVEWLLSA-NH₂ (SEQ ID NO;

126) and insulin lispro, H—

H-Aib-QGTFTSDYSKYLDS-K(Hexadecanoyl-isoGlu)-AAHDFVEWLLSA-NH₂ (SEQ ID NO;

126) and degludec, H—

H-Aib-QGTFTSDYSKYLDS-K(Hexadecanoyl-isoGlu)-AAHDFVEWLLSA-NH₂ (SEQ ID NO;

126) and Actraphane, H—

H-Aib-QGTFTSDYSKYLDS-K(Hexadecanoyl-isoGlu)-AAHDFVEWLLSA-NH₂ (SEQ ID NO;

126) and LY2963016, H—

H-Aib-QGTFTSDYSKYLDS-K(Hexadecanoyl-isoGlu)-AAHDFVEWLLSA-NH₂ (SEQ ID NO;

1261 and LY2605541, H—

H-Aib-QGTFTSDYSKYLDS-K(Hexadecanoyl-isoGlu)-AAHDFVEWLLSA-NH₂ (SEQ ID NO;

126) and pegylated insulin Lispro, and H—

H-Aib-QGTFTSDYSKYLDS-K(Hexadecanoyl-isoGlu)-AAHDFVEWLLSA-NH₂ (SEQ ID NO;

126) and NN-1953, IN-105, BIOD-620 and Analog-PH20.

93. The method of claim 5, wherein said compound having the formula R¹—

Z—

R² is administered in a dosage selected from the group consisting of 0.1 nmol/kg to 1 μ

mol/kg and 3 nmol/kg to 30 nmol/kg.

95. The method of claim 5, wherein said insulin analogue is administered in a dosage selected from the group consisting of 0.02 U/kg to 20 U/kg, 0.1 U/kg to 0.3 U/kg, and about 0.2 U/kg.

99. The method of claim 5, wherein said compound or combination of compounds is administered in an amount selected from:

an amount sufficient to reduce food intake in said subject by at least 5%, 10%, 15%, 20%, 25%, 30%, or 50%,

an amount sufficient to reduce the subject'"'"'s fasting blood glucose level by at least 1, 2, 3, 4, 5, 6, 8, 10, 11, 12, 15, or 20 mM,

an amount sufficient to reduce the subjects HbA1c level by at least 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.8%, 1.0%, 1.5%, or 2.0%,

an amount which results in a body weight reduction of at least 3%, 5%, 8%, 10%, 12%, 15% or 20% within 1 year of starting administration,

an amount which results in a body weight reduction of at least 1%, 2%, 3%, 4%, 5%, 6%, 8%, or 10%, 15% within six months of starting administration, and

an amount which results in a body weight reduction of at least 0.5%, 1%, 2%, 3%, 4%, 5%, 6%, 8%, 10% or 15% within three months of starting administration.

112. The method of claim 5, wherein the compound (a) an insulin analogue (b) are formulated for simultaneous or sequential administration.

113. The method of claim 5, wherein the compound (a) and insulin analogue (b) are formulated as separate medicaments.

114. The method of claim 5, wherein said subject is a human.

7-8. -8. (canceled)

10-11. -11. (canceled)

20-22. -22. (canceled)

25. (canceled)

27-28. -28. (canceled)

30-34. -34. (canceled)

36. (canceled)

38. (canceled)

40. (canceled)

43-70. -70. (canceled)

72-92. -92. (canceled)

94. (canceled)

96-98. -98. (canceled)

100-108. -108. (canceled)

109. A kit comprising:

(a) 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;

His-X2-Gln-Gly-Thr-Phe-Thr-Ser-Asp-Tyr-Ser-X12-Tyr-Leu-Asp-X16-X17-Ala-Ala-X20-X21-Phe-Val-X24-Trp-Leu-X27-X28-Ala-X30;

 

 

(I)

wherein

X2 is selected from Aib and Ser;

X12 is selected from Lys, Arg, or Leu;

X16 is selected from Arg and X;

X17 is selected from Arg and X;

X20 is selected from Arg, His, and X;

X21 is selected from Asp and Glu;

X24 is selected from Ala and X;

X27 is selected from Leu and X;

X28 is selected from Arg and X;

X30 is X or is absent;

wherein at least one of X16, X17, X20, X24, X27, X28, and X30 is X;

and wherein each residue X is independently selected from the group consisting of Glu, Lys, Ser, Cys, Dbu, Dpr, and Orn;

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; and

(b) an insulin analogue.

111. The kit of claim 109, wherein said insulin analogue is selected from the group consisting of insulin glulisine, insulin lispro, Degludec, Actraphane HM, LY2963016, LY2605541, pegylated insulin Lispro, insulin glargine (Lantus) insulin detemir (Levemir), insulin isophane, insulin aspart, insulin buccal, hyaluronidase insulin, insulin protamine, NN-1953, IN-105, BIOD-620 and Analog-PH20.

110. (canceled)

115. A method for preventing or reducing weight gain;

promoting weight loss;

improving circulating glucose levels, glucose tolerance or circulating cholesterol levels;

lowering circulating LDL levels;

increasing HDL/LDL ratio;

lowering circulating triacylglycerol levels, lowering circulating free fatty acids or treating a condition caused or characterized by excess body weight in a mammalian subject that is receiving an insulin analogue, said method comprising administering to said subject in an effective amount 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;

His-X2-Gln-Gly-Thr-Phe-Thr-Ser-Asp-Tyr-Ser-X12-Tyr-Leu-Asp-X16-X17-Ala-Ala-X20-X21-Phe-Val-X24-Trp-Leu-X27-X28-Ala-X30;

 

 

(I)

wherein

X2 is selected from Aib and Ser;

X12 is selected from Lys, Arg, or Leu;

X16 is selected from Arg and X;

X17 is selected from Arg and X;

X20 is selected from Arg, His, and X;

X21 is selected from Asp and Glu;

X24 is selected from Ala and X;

X27 is selected from Leu and X;

X28 is selected from Arg and X;

X30 is X or is absent;

wherein at least one of X16, X17, X20, X24, X27, X28, and X30 is X;

and wherein each residue X is independently selected from the group consisting of Glu, Lys, Ser, Cys, Dbu, Dpr, and Orn;

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.

116. The method of claim 115, wherein said insulin analogue is selected from the group consisting of insulin glulisine (Apidra), insulin lispro (Humalog), Degludec, Actraphane HM, LY2963016, LY2605541, pegylated insulin Lispro, insulin glargine (Lantus), insulin detemir (Levemir), insulin isophane, insulin aspart, insulin buccal, hyaluronidase insulin, insulin protamine, NN-1953, IN-105, BIOD-620 and Analog-PH20.