A-C-B proinsulin, method of manufacturing and using same, and intermediates in insulin production

• US 5,304,473 A
• Filed: 06/11/1991
• Issued: 04/19/1994
• Est. Priority Date: 06/11/1991
• Status: Expired due to Fees

• Current Assignee: Eli Lilly and Company
• Original Assignee: Eli Lilly and Company
• Inventors: DiMarchi, Richard D., Belagaje, Rama M., Heath, William F. Jr., Long, Harlan B.
• Primary Examiner(s): Lacey, David L.
• Assistant Examiner(s): Adams, Donald E.
• Application Number:
  US07/715,183
• Time in Patent Office: 1,043 Days
• Field of Search: 435/91, 435/69.7, 435/252.33, 514/3, 530/350, 530/303, 536/27, 536/23.51
• US Class Current: 435/69.7
• 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: A61P 3/08 for glucose homeostasis pancreatic hormones A61P5/48
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: C07K 14/62 Insulins

Abstract

The instant invention provides novel molecules derived from the components of proinsulin using recombinant DNA technology. The invention provides molecules of the formula A--C--B wherein A is the A-chain of an insulin species, B is the B-chain of an insulin species and C is a connecting peptide. These molecules possess insulin-like activity and are useful for the treatment of diabetes mellitus, particularly non-insulin dependent diabetes mellitus. These molecules are also useful for the production of insulin and constitute a novel pathway for the recombinant production of insulin species. The invention provides a method of making insulin proceeding through the compounds of the invention as intermediates. The instant invention further provides recombinant DNA compounds which encode the compounds of the invention.

Claims

1. A Polypeptide compound of the formula:

space="preserve" listing-type="equation">Met.sub.x --A--C--B

wherein;

Met=the amino acid methionine,

x=0 or 1,

A=the A chain of insulin or a functional derivative thereof,

B=the B chain of insulin or a functional derivative thereof,

C=the C peptide of insulin or a peptide of the formula;

space="preserve" listing-type="equation">X.sub.1 --X.sub.2 --P--X.sub.3 --X.sub.4,

wherein;

X₁, X₂, X₃, and X₄ are basic amino acids,

X₁, X₂, X₃, and X₄ are the same or different, and,

P is a peptide of from 4 to 35 amino acids which does not contain a cysteine residue,

and pharmaceutically acceptable salts thereof.

2. The compound of claim 1 wherein the amino acid at position 21 of the A-chain is selected from the groups consisting of Gly, Ala, Asp or Asn.

3. The compound of claim 2 wherein the amino acid at position 10 of the B-chain is Asp.

4. The compound of claim 3 wherein the amino acid at position 30 of the B-chain is Ala or deleted.

5. The compound of claim 3 wherein the amino acid at position 29 of the B-chain is Pro.

6. The compound of claim 5 wherein the amino acid at position 28 of the B-chain is Glu or Lys.

7. The compound of claim 2 wherein the amino acid at position 27 of the B-chain is Arg.

8. The compound of claim 7 wherein the amino acid at position 13 of the B-chain is Gln.

9. The compound of claim 7 wherein the amino acid at position 17 of the A-chain is Glu.

10. The compound of claim 7 wherein the amino acid at position 21 of the A-chain is Gly and the amino acid at position 30 of the B-chain is Thr-NH₂.

11. The compound of claim 2 wherein the amino acid at position 29 of the B-chain in Pro.

12. The compound of claim 11 wherein the amino acid at position 28 of the B-chain is Lys or Glu.

13. The compound of claim 12 wherein the amino acid at position 30 of the B-chain is Ala.

14. The compound of claim 2 wherein the amino acid at position 30 of the B-chain is Ala.

15. The compound of claim 1 wherein the amino acid at position 10 of the B-chain is Asp or His.

16. The compound of claim 15 wherein the amino acid at position 28 of the B-chain is Asp.

17. The compound of claim 15 wherein the amino acid at position 30 of the B-chain is deleted.

18. The compound of claim 1 wherein the amino acid at position 28 of the B-chain is Lys, and the amino acid at position 29 of the B-chain is Pro.

19. The compound of claim 1 wherein the amino acids at positions 29 and 30 of the B-chain are deleted.

20. A DNA compound encoding the compound of claim 1.

21. A DNA compound encoding the compound of claim 2.

22. A DNA compound encoding the compound of claim 3.

23. A DNA compound encoding the compound of claim 4.

24. A DNA compound encoding the compound of claim 5.

25. A DNA compound encoding the compound of claim 6.

26. A DNA compound encoding the compound of claim 7.

27. A DNA compound encoding the compound of claim 8.

28. A DNA compound encoding the compound of claim 9.

29. A DNA compound encoding the compound of claim 10.

30. A DNA compound encoding the compound of claim 11.

31. A DNA compound encoding the compound of claim 12.

32. A DNA compound encoding the compound of claim 13.

33. A DNA compound encoding the compound of claim 14.

34. A DNA compound encoding the compound of claim 15.

35. A DNA compound encoding the compound of claim 16.

36. A DNA compound encoding the compound of claim 17.

37. A DNA compound encoding the compound of claim 18.

38. A DNA compound encoding the compound of claim 19.

39. A method of treating diabetes by administering a pharmaceutically acceptable dose of the compound of claim 1.

41. A method to stimulate smooth muscle cell proliferation by administering the compound of claim 1.

40. A method of treating non-insulin dependent diabetes mellitus by administering a pharmaceutically acceptable dose of the compound of formula 1.