Glucagon analogs exhibiting enhanced solubility in physiological pH buffers

• US 8,669,228 B2
• Filed: 01/03/2008
• Issued: 03/11/2014
• Est. Priority Date: 01/05/2007
• Status: Expired due to Fees

• Current Assignee: Indiana University Research & Technology Corporation (Indiana University)
• Original Assignee: Indiana University Research & Technology Corporation (Indiana University)
• Inventors: Dimarchi, Richard D., Dimarchi, Maria, Chabenne, Joseph
• Primary Examiner(s): Saoud, Christine J
• Application Number:
  US12/522,129
• Publication Number:
  US 20100190699A1
• Time in Patent Office: 2,259 Days
• Field of Search: None
• US Class Current: 514/7.2
• CPC Class Codes: A61P 1/00 Drugs for disorders of the alimentary tract or the digestive system
• 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 3/08 for glucose homeostasis pancreatic hormones A61P5/48
• CPC Class Codes: C07K 14/605 Glucagons

Abstract

Modified glucagon peptides are disclosed having improved solubility while retaining glucagon agonist activity. The glycogen peptides have been modified by substitution of native amino acids with, and/or addition of, charged amino acids to the carboxy terminus of the peptide. The modified glucagon agonists can be further modified by pegylation, or the addition of a carboxy terminal peptide selected from the group consisting of SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 23, or both to further enhance the solubility of the glucagon agonist analogs.

Claims

1. A glucagon peptide with improved solubility relative to native glucagon (SEQ ID NO:

1), that is an analog of SEQ ID NO;

1 comprising (a) one, two, or three charged amino acid(s) C-terminal to the amino acid at position 27 of the glucagon peptide, (b) a Leu or Nle at position 27, and (c) up to 4 additional amino acid modifications relative to SEQ ID NO;

1, wherein the glucagon peptide retains improved solubility and glucagon activity.

2. The glucagon peptide of claim 1, wherein said one, two, or three charged amino acid are negatively charged amino acids.

3. The glucagon peptide of claim 2, wherein the negatively charged amino acids are selected from the group consisting of Glu and Asp.

4. The glucagon peptide of claim 3, wherein (i) the amino acid at position 28 is Asp, or (ii) the amino acid at position 29 is Glu, or both (i) and (ii).

5. The glucagon peptide of claim 4, wherein the additional amino acid modifications are:

amino acid substitutions at positions 1, 2, 5, 7, 10, 11, 12, 13, 14, 16, 17, 18, 19, 20, 21, and 24 relative to SEQ ID NO;

1.

6. A homo dimer comprising two glucagon peptides of claim 5 bound to one another through a linker.

7. The glucagon peptide of claim 2, wherein two or more charged amino acids C-terminal to the amino acid at position 27 are negatively charged.

8. The glucagon peptide of claim 7, comprising

a. a substitution of the amino acid at position 15 with Glu, homo-Glu, cysteic acid, homo cysteic acid;

b. a hydrophilic moiety covalently bound at position 16, 17, 20, 21, 24 or 29 of said glucagon peptide;

c. a sequence selected from the group consisting of SEQ ID NO;

20, SEQ ID NO;

21, and SEQ ID NO;

22 bound to the carboxy terminus of said glucagon peptide;

d. an amino acid at position 2 selected from the group consisting of d-serine, alanine, glycine, n-methyl serine and amino isobutyric acid;

or the amino acid at position 1 is selected from the group consisting of dhistidine, desaminohistidine, hydroxyl-histidine, acetyl-histidine and homo-histidine;

or

e. a combination thereof.

10. The glucagon peptide of claim 1 wherein the glucagon peptide comprises the sequence of any one of SEQ ID NOs:

7, 8, 9, 11, 12, 13, 32, 33, 34, or 38.

11. A pharmaceutical composition comprising the glucagon peptide of claim 5, 9, or 7 or a pharmaceutically acceptable salt thereof;

and a pharmaceutically acceptable carrier.

15. A solution comprising the glucagon peptide of claim 1, 5, or 9 or a pharmaceutically acceptable salt thereof;

and a pharmaceutically acceptable carrier;

said solution at pH of about 5.5 to about 8.0.

16. A kit for administering a glucagon agonist to a patient in need thereof, said kit comprising the solution of claim 15;

and a device for administering said composition to a patient.

17. The kit of claim 16 wherein said device:

is an aerosol dispensing device, wherein the composition is prepackaged within the aerosol device;

or

comprises a syringe and a needle, wherein the composition is prepackaged within the syringe.

18. A method of treating hypoglycemia in a patient, comprising administering to the patient the glucagon peptide of claim 1, 5, or 9 in an amount effective for treating hypoglycemia.

9. A glucagon peptide with improved solubility relative to native glucagon (SEQ ID NO:

1) comprising the sequence of SEQ ID NO;

33 or an analog of SEQ ID NO;

33, wherein said analog differs from SEQ ID NO;

33 by one to three amino acid substitutions, said amino acid substitutions located at one or more positions selected from positions 1, 2, 5, 7, 10, 11, 12, 13, 14, 16, 17, 18, 19, 20, 21 and 24 of the analog, with the proviso that said glucagon peptide comprises a charged amino acid at position 28 or 29, or an additional acidic amino acid added to the carboxy terminus of the peptide.

12. The glucagon peptide of claim 9 further comprising a hydrophilic moiety covalently bound at position 16, 17, 21 or 24 of the glucagon peptide.

13. The glucagon peptide of claim 12 wherein the hydrophilic moiety is polyethylene glycol (PEG).

14. The glucagon peptide of claim 13 wherein the glucagon peptide further comprises a sequence selected from the group consisting of SEQ ID NO:

20, SEQ ID NO;

21, and SEQ ID NO;

22 bound to the carboxy terminus of said glucagon peptide.