Peptide compounds exhibiting glucagon antagonist and GLP-1 agonist activity

• US 8,980,830 B2
• Filed: 10/27/2008
• Issued: 03/17/2015
• Est. Priority Date: 10/30/2007
• Status: Active Grant

• Current Assignee: Indiana University Research & Technology Corporation (Indiana University)
• Original Assignee: Indiana University Research & Technology Corporation (Indiana University)
• Inventors: Dimarchi, Richard D., Yang, Bin, Ouyang, Chenguang
• Primary Examiner(s): Orwig, Kevin S
• Application Number:
  US12/740,206
• Publication Number:
  US 20110098217A1
• Time in Patent Office: 2,332 Days
• Field of Search: None
• US Class Current: 514/6.9
• 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 38/16 Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof enzyme inhibitors A61K38/005
• CPC Class Codes: A61K 38/22 Hormones derived from pro-opiomelanocortin, pro-enkephalin or pro-dynorphin A61K38/33, e.g. corticotropin A61K38/35
• CPC Class Codes: A61K 38/26 Glucagons
• CPC Class Codes: A61P 1/00 Drugs for disorders of the alimentary tract or the digestive system
• CPC Class Codes: A61P 13/12 of the kidneys
• CPC Class Codes: A61P 19/00 Drugs for skeletal disorders
• CPC Class Codes: A61P 21/00 Drugs for disorders of the muscular or neuromuscular system
• CPC Class Codes: A61P 27/02 Ophthalmic agents
• 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 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• 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
• CPC Class Codes: C07K 7/02 Linear peptides containing at least one abnormal peptide link

Abstract

Glucagon analogs are disclosed that exhibit both glucagon antagonist and GLP-1 agonist activity. In one embodiment, the glucagon antagonist/GLP-1 agonist comprises a modified amino acid sequence of native glucagon, in which the first one to five N-terminal amino acids of native glucagon is deleted and in which the alpha helix is stabilized.

Claims

1. A glucagon antagonist/GLP-1 agonist comprising the sequence of SEQ ID NO:

51, wherein the amino acids at positions 4 and 7, positions 7 and 11, positions 11 and 15, positions 15 and 19, or positions 19 and 23 of SEQ ID NO;

51 are linked via a lactam bridge, or an oxy derivative of the glucagon antagonist/GLP-1 agonist.

2. A glucagon antagonist/GLP-1 agonist peptide comprising an intramolecular bridge formed between two amino acid side chains of the glucagon peptide and an amino acid sequence of SEQ ID NO:

51 or a derivative thereof wherein the derivative differs from the peptide of SEQ ID NO;

51 by amino acid substitutions at one to three amino acid positions selected from positions 1, 5, 6, 8, 9, 12, 13 and 14, said peptide exhibiting at least 90% of the maximum agonism of native GLP-1 at the GLP-1 receptor, and exhibiting glucagon antagonist activity, that reduces the maximum glucagon-induced cAMP production by the glucagon receptor by at least about 80%, as measured by cAMP production in an in vitro assay.

3. A peptide comprising (1) an intramolecular bridge, or an alpha, alpha-di-substituted amino acid, or an acidic amino acid at position 16 (according to the numbering of SEQ ID NO:

1), or a combination thereof, (2) a C-terminal amide or ester in place of a C-terminal carboxylate, and (3) a general structure of A-B-C,

wherein A is selected from the group consisting of

(i) phenyl lactic acid (PLA);

(ii) an oxy derivative of PLA; and

(iii) a peptide of 2 to 6 amino acids in which two consecutive amino acids of the peptide are linked via an ester or ether bond;

wherein B represents amino acids p to 26 of SEQ ID NO;

1, wherein p is 3, 4, 5, 6, or 7, optionally comprising one or more amino acid modifications selected from the group consisting of;

(iv) Asp at position 9 (according to the amino acid numbering of SEQ ID NO;

     1) is substituted with a Glu, a sulfonic acid derivative of Cys, homoglutamic acid, β

-homoglutamic acid, or an alkylcarboxylate derivative of cysteine having the structure of;

4. The peptide of claim 3, wherein the oxy derivative of PLA is a depsipeptide comprising PLA covalently linked via an ester bond to an amino acid or a peptide.

5. The peptide of claim 4, wherein the depsipeptide comprises PLA covalently linked via an ester bond to Xaa₃, or to a peptide Xaa₂-Xaa₃ or Xaa₁-Xaa₂-Xaa₃, wherein Xaa₃ is Gln or Glu, Xaa₂ is selected from a group consisting of:

Ser, D-serine, D-alanine, valine, glycine, N-methyl serine, N-methyl alanine, and aminoisobutyric acid (AIB); and

Xaa₁ is selected from a group consisting of;

His, D-histidine, alpha, alpha-dimethyl imidiazole acetic acid (DMIA), N-methyl histidine, alpha-methyl histidine, imidazole acetic acid, desaminohistidine, hydroxyl-histidine, acetyl-histidine and homo-histidine.

6. The peptide of claim 3, wherein B represents amino acids 7 to 26 of SEQ ID NO:

1.

7. The peptide of claim 3, wherein B comprises the amino acid modification designated as (v) or (vi), or a combination thereof.

8. The peptide of claim 7, wherein B further comprises one or more amino acid modifications selected from the group consisting of (iv), (vii), (viii), (ix), (x), and a combination thereof.

9. The peptide of claim 3, comprising a hydrophilic moiety covalently bound to an amino acid residue at position 16, 21, or 24 according to the amino acid numbering of SEQ ID NO:

1, or the C-terminal residue of the peptide.

10. The peptide of claim 3, comprising an amino acid covalently attached to an acyl group or alkyl group via an ester, ether, thioether, amide, or alkyl amine linkage, wherein the amino acid is at position 10, 20, or 24 (according to the amino acid numbering of SEQ ID NO:

1).

11. The peptide of claim 3, wherein the intramolecular bridge is a lactam bridge or a salt bridge.

12. The peptide of claim 11, comprising a lactam bridge between the amino acids at positions 9 and 12, the amino acids at positions 12 and 16, the amino acids at positions 16 and 20, the amino acids at positions 20 and 24, or the amino acids at positions 24 and 28 (according to the amino acid numbering of SEQ ID NO:

1).

13. The peptide of claim 3, comprising AIB at position 16, 20, 21, or 24 (according to the amino acid numbering of SEQ ID NO:

1).

14. The peptide of claim 3, comprising the amino acid sequence of any of SEQ ID NOs:

60-70, 73-78, 80-88, 90-96, 103, 104, 106, and 114-118, or comprising the amino acid sequence of any of Peptides 2-6 of Table 13, Peptides 1-8 of Table 14, and Peptides 2-6, 8, and 9 of Table 15.

15. A multimer or dimer comprising a peptide of claim 3.

16. A conjugate comprising a peptide of claim 3 and a heterologous peptide or polypeptide.

17. A pharmaceutical composition comprising a peptide of claim 3, and a pharmaceutically acceptable carrier.

18. A method of treating hyperglycemia or diabetes in a patient in need thereof, comprising administering to the patient the pharmaceutical composition of claim 17 in an amount effective to treat the hyperglycemia or diabetes.

19. A method of suppressing appetite, reducing weight gain, or inducing weight loss in a patient in need thereof, comprising administering to the patient the pharmaceutical composition of claim 17 in an amount effective to suppress appetite, reduce weight gain, or induce weight loss.

20. A kit comprising a peptide of claim 3 and a set of user instructions.

21. The peptide of claim 14, comprising the amino acid sequence of SEQ ID NO:

76.

22. The peptide of claim 21, comprising a hydrophilic moiety covalently bound to an amino acid residue at position 16, 21, or 24 according to the amino acid numbering of SEQ ID NO:

1, or the C-terminal residue of the peptide, wherein the hydrophilic moiety is a polyethylene glycol chain.