Amide based glucagon and superfamily peptide prodrugs

• US 8,969,288 B2
• Filed: 12/18/2009
• Issued: 03/03/2015
• Est. Priority Date: 12/19/2008
• 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., Kou, Binbin
• Primary Examiner(s): Skowronek, Karlheinz R
• Assistant Examiner(s): MADER, CATHERINE J
• Application Number:
  US13/130,962
• Publication Number:
  US 20110288003A1
• Time in Patent Office: 1,901 Days
• Field of Search: 514/1.3
• US Class Current: 514/1.3
• 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: A61P 1/00 Drugs for disorders of the alimentary tract or the digestive system
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous 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 3/06 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 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00

Abstract

Prodrug formulations of glucagon superfamily peptides are provided wherein the glucagon superfamily peptide has been modified by the linkage of a dipeptide to the glucagon superfamily through an amide bond linkage. The prodrugs disclosed herein have extended half lives and are converted to the active form at physiological conditions through a non-enzymatic reaction driven by chemical instability.

Claims

1. A complex comprising the structure:

 

A-B-Q;

wherein Q is a glucagon superfamily peptide;

A is an alpha-amino acid or an alpha-hydroxy acid, wherein when the alpha-carbon is chiral, A is in the D-stereochemical configuration;

B is an N-alkylated amino acid linked to Q through an amide bond between A-B and a primary amine of Q;

wherein A, B, or the amino acid of Q to which A-B is linked is a non-coded amino acid, further wherein the chemical cleavage half-life (t_1/2) of A-B from Q is at least about 1 hour to about 1 week in PBS under physiological conditions.

2. The complex of claim 1, wherein the cleavage half-life of A-B from Q in PBS under physiological conditions, is not more than two fold the cleavage half-life of A-B from Q in a solution comprising a DPP-IV protease, wherein the solution comprising a DPP-IV protease is serum.

6. The complex of claim 1, wherein said primary amine is the alpha amino group of the N-terminal amino acid of Q.

7. The complex of claim 6, wherein A-B comprises the structure:

8. The complex of claim 6, wherein A-B comprises a structure of:

9. The complex of claim 6, wherein A-B comprises a structure of:

10. The complex of claim 6, wherein A-B comprises a structure of:

11. The complex of claim 1, wherein said primary amine is a side chain amino group of an internal amino acid of Q, said internal amino acid comprising the structure of Formula V:

12. The complex of claim 11, wherein A-B comprises the structure:

13. The complex of claim 11, wherein A-B comprises a structure of:

14. The complex of claim 11, wherein A-B comprises a structure of:

15. The complex of claim 11, wherein the amino acid of Formula V is located at a position of Q corresponding to position 12, 16, 17, 18, 20, 28, or 29 of native glucagon (SEQ ID NO:

701).

16. The complex of claim 1, wherein said primary amine is a substituent on an aryl of an aromatic amino acid of Q.

17. The complex of claim 16, wherein said aromatic amino acid (i) is an internal amino acid of Q, (ii) is located at a position of Q corresponding to position 10, 13, 22, or 25 of native glucagon (SEQ ID NO:

701), (iii) is selected from the group consisting of amino-Phe, amino-napthyl alanine, amino tryptophan, amino-phenyl-glycine, amino-homo-Phe, and amino tyrosine, or (iv) comprises the structure of Formula III;

18. The complex of claim 16, wherein A-B comprises a structure of:

19. The complex of claim 16, wherein A-B comprises a structure of:

20. The complex of claim 16, wherein A-B comprises a structure of:

21. The complex of claim 1, further comprising a hydrophilic moiety covalently linked to the complex or a non-native acyl group or a non-native alkyl group covalently linked to an amino acid side chain.

22. The complex of claim 1, wherein Q comprises an amino acid sequence at least 50% identical to native glucagon (SEQ ID NO:

701) that retains the alpha-helix conformation of the amino acids corresponding to amino acids 12-29 of SEQ ID NO;

701.

23. The complex of claim 1, wherein Q is selected from the group consisting of SEQ ID NOs:

1-564, 566-570, 573-575, 577, 579-580, 585-612, 616, 618-632, 634-642, 647, 657-684, 701-732, 801-878, 883-919, 1001-1262, 1301-1371, 1401-1518, 17011708, 1710, 1711, 1731-1734, 1738, 1740, 1741, 1745 and 1747-1776.

24. A sterile pharmaceutical composition comprising the complex of claim 1, and a pharmaceutically acceptable carrier.

25. The complex of claim 21, wherein said acyl group or alkyl group is covalently linked to A-B.

26. The complex of claim 25, wherein said acyl group is a C4to C30 fatty acyl group.

27. The complex of claim 1, wherein Q comprises the amino acid sequence of native glucagon (SEQ ID NO:

701) with up to 10 amino acid modifications relative to the native glucagon sequence.

28. The complex of claim 23, wherein Q comprises the sequence of SEQ ID NO:

251.

3. A complex comprising the structure A-B-Q;

wherein Q is a glucagon superfamily peptide and wherein A-B comprises the structure;

4. The complex of claim 3 wherein the glucagon superfamily peptide is a glucagon related peptide.

5. The complex of claim 3 wherein

(i) R₁ and R₈ are independently H or C₁-C₈ alkyl;

R₂ and R₄ are independently selected from the group consisting of H, C₁-C₈ alkyl, C₂-C₈ alkenyl, (C₁-C₄ alkyl)OH, (C₁-C₄ alkyl)SH, (C₂-C₃ alkyl)SCH₃, (C₁-C₄ alkyl)CONH₂, (C₁-C₄ alkyl)COOH, (C₁-C₄ alkyl)NH₂, (C₁-C₄ alkyl)NHC(NH₂+) NH₂, (C₀-C₄ alkyl)(C₃-C₆ cycloalkyl), (C₀-C₄ alkyl)(C₂-C₅ heterocyclic), (C₀-C₄ alkyl)(C₆-C₁₀ aryl)R₇, and CH₂(C₃-C₉ heteroaryl), or R₁ and R₂ together with the atoms to which they are attached form a C₃-C₁₂ cycloalkyl or aryl;

R₅ is NHR₆; and

R₆ is H or C₁-C₈ alkyl, or

(ii) R₁ and R₈ are independently H or C₁-C₈ alkyl;

R₂ and R₄ are independently selected from the group consisting of H, C₁-C₈ alkyl, C₂-C₈ alkenyl, (C₁-C₄ alkyl)OH, (C₁-C₄ alkyl)SH, (C₂-C₃ alkyl)SCH₃, (C₁-C₄ alkyl)CONH₂, (C₁-C₄ alkyl)COOH, (C₁-C₄ alkyl)NH₂, (C₁-C₄ alkyl)NHC(NH₂+) NH₂, (C₀-C₄ alkyl)(C₃-C₆ cycloalkyl), (C₀-C₄ alkyl)(C₂-C₅ heterocyclic), (C₀-C₄ alkyl)(C₆-C₁₀ aryl)R₇, and CH₂(C₃-C₉ heteroaryl), or R₁ and R₂ together with the atoms to which they are attached form a C₃-C₁₂ cycloalkyl;

R₃ is C₁-C₁₈ alkyl;

R₅ is NHR₆;

R₆ is H or C₁-C₈ alkyl; and

R₇ is selected from the group consisting of hydrogen, C₁-C₁₈ alkyl, C₂-C₁₈ alkenyl, (C₀-C₄ alkyl)CONH₂, (C₀-C₄ alkyl)COOH, (C₀-C₄ alkyl)NH₂, (C₀-C₄ alkyl)OH, and halo.

29. The complex of claim 3, wherein R₁ is selected from the group consisting of C₁-C₁₈ alkyl, C₂-C₁₈ alkenyl, (C₁-C₁₈ alkyl)OH, (C₁-C₁₈ alkyl)SH, (C₂-C₃ alkyl)SCH₃, (C₁-C₄ alkyl)CONH₂, (C₁-C₄ alkyl)COOH, (C₁-C₄ alkyl)NH₂, (C₁-C₄ alkyl)NHC(NH₂⁺)NH₂, (C₀-C₄ alkyl)(C₃-C₆ cycloalkyl), (C₀-C₄ alkyl)(C₂-C₅ heterocyclic), (C₀-C₄ alkyl)(C₆-C₁₀ aryl)R₇, (C₁-C₄ alkyl)(C₃-C₉ heteroaryl), and C₁-C₁₂ alkyl(W₁)C₁-C₁₂ alkyl, wherein W₁ is a heteroatom selected from the group consisting of N, S and O, or R₁ and R₂ together with the atoms to which they are attached form a C₃-C₁₂ cycloalkyl or aryl.