Insulin derivatives

• US 3,950,517 A
• Filed: 12/06/1974
• Issued: 04/13/1976
• Est. Priority Date: 05/08/1970
• Status: Expired due to Term

• Current Assignee: National Research Development Corporation
• Original Assignee: National Research Development Corporation
• Inventors: Shall, Sydney, Lindsay, David Gordon
• Primary Examiner(s): Waddell, Frederick E.
• Application Number:
  US05/530,143
• Time in Patent Office: 494 Days
• Field of Search: 424/178
• US Class Current: 514/6.3
• 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: C07K 14/62 Insulins

Abstract

A physiologically acceptable insulin derivative has the terminal amino group of the B chain (B₁, phenylalanine) protected by an acyl group or other blocking group containing up to 7 carbon atoms and the amino group of the A chain (A₁, glycine) is either free or protected by means of an acyl or other blocking group containing no more than four carbon atoms and preferably no more than three atoms other than hydrogen. Pharmaceutical preparations may be prepared containing an effective amount of such an insulin derivative together with a physiologically acceptable diluent. An insulin derivative of reduced antigenicity is obtainable from porcine or bovine insulin by reacting the insulin with an acylating agent or other blocking reagent to acylate or otherwise block the amino group of the B₁ amino acid and optionally block the A₁ and/or B₂₉ amino acid, followed by purification of the derivative by various methods.

Claims

1. A pharmaceutical composition useful for treating diabetes in humans which comprises a hypoglycemically effective amount of a mono-, di- or tri-substituted insulin in which the terminal amino group of the B chain (B₁ phenylalanine) is protected by an acyl or other blocking substituent containing up to 7 carbon atoms selected from the group consisting of formyl, acetyl, trifluoroacetyl, cyclopropane-carbonyl, aceto-acetyl, benzoyl, 2,2-dimethyl-3-formyl-L-thiazolidine-4-carbonyl, carbamyl, methylcarbamyl, thiocarbamyl, methylthiocarbamyl, ##EQU4## HOCH₂ CH₂ CO--, HOOCCH₂ CO-- and H₂ NOCCH₂ CO--, the terminal amino group of the A chain (A₁ glycine) is either free or protected by an acyl or other blocking substituent containing no more than 4 carbon atoms and no free primary amino group selected from the group consisting of formyl, acetyl, trifluoroacetyl, cyclopropane-carbonyl, acetoacetyl, carbamyl, methylcarbamyl, thiocarbamyl, methylthiocarbamyl, ##EQU5## HOCH₂ CH₂ CO--, HOOCCH₂ CO-- and H₂ NOC--CH₂ CO-- and the amino group of the B₂₉ amino acid (lysine) is either free or protected by an acyl or other blocking substituent containing no more than 4 carbon atoms and no free primary amino group selected from the group consisting of formyl, acetyl, trifluoroacetyl, cyclopropanecarbonyl, acetoacetyl, carbamyl, methylcarbamyl, thiocarbamyl, methylthiocarbamyl, ##EQU6## HOCH₂ CH₂ CO--, HOOC--CH₂ --CO-- and H₂ NOC--CH₂ CO-- in combination with a pharmaceutically acceptable diluent.

2. A composition according to claim 1 in which the terminal amino group of the A chain (A₁ glycine) is either free or protected by an acyl or other blocking substituent containing no more than three atoms other than hydrogen selected from the group consisting of formyl, acetyl, carbamyl, thiocarbamyl, ##EQU7## and the amino group of the B₂₉ amino acid (lysine) is either free or protected by an acyl or other blocking substituent containing no more than three atoms other than hydrogen selected from the group consisting of formyl, acetyl, carbamyl, thiocarbamyl, ##EQU8##

3. A composition according to claim 1 in which the B₁ amino acid is protected by acetyl or acetoacetyl.

4. A composition according to claim 1 in which the B₁ amino acid is protected by 2,2-dimethyl-3-formyl-L-thiazolidine-4-carbonyl.

5. A composition according to claim 1 in which at least one of the substituents is a carbamyl group.

6. A composition according to claim 1 wherein the insulin is mono-substituted.

7. A composition according to claim 1 wherein the insulin is di-substituted.

8. A composition according to claim 1 wherein the insulin is tri-substituted.

9. A composition according to claim 1 wherein the A₁ and B₁ protective substituents are the same.

10. A composition according to claim 1 wherein the insulin is B₁ (phenylalanine)-N-monocarbamyl insulin.

11. A composition according to claim 1 wherein the insulin is A₁ (glycine) B₁ (phenylalanine)-N,N'"'"'-dicarbamyl insulin.

12. A composition according to claim 1 wherein the insulin is A₁ (glycine)B₁ (phenylalanine)B₂₉ (lysine)-N,N'"'"',N"-tricarbamyl insulin.

13. A pharmaceutical composition according to claim 1 in injectable form.

14. A composition according to claim 8 wherein the substituents are the same.

28. A method according to claim 13 wherein the substituents are the same.

15. A pharmaceutical composition useful for treating diabetes in humans which comprises a hypoglycemically effective amount of a mixture of A₁ (glycine), B₁ (phenylalanine), B₂₉ (lysine)-N,N'"'"',N"-tricarbamyl insulin and A₁ (glycine), B₁ (phenylalanine)-N,N'"'"'-dicarbamyl insulin in combination with a pharmaceutically acceptable diluent.

16. A method of treating diabetes in humans which comprises parenterally administering to such human a hypoglycemically effective amount of a mono-, di- or tri-substituted insulin in which the terminal amino group of the B chain (B₁ phenylalanine) is protected by an acyl or other blocking substituent containing up to 7 carbon atoms selected from the group consisting of formyl, acetyl, trifluoroacetyl, cyclopropane-carbonyl, aceto-acetyl, benzoyl, 2,2-dimethyl-3-formyl-L-thiazolidine-4-carbonyl, carbamyl, methylcarbamyl, thiocarbamyl, methylthiocarbamyl, ##EQU9## HOCH₂ CH₂ CO--, HOOCCH₂ CO-- and H₂ NOCCH₂ CO--, the terminal amino group of the A chain (A₁ glycine) is either free or protected by an acyl or other blocking substituent containing no more than 4 carbon atoms and no free primary amino group selected from the group consisting of formyl, acetyl, trifluoroacetyl, cyclopropane-carbonyl, acetoacetyl, carbamyl, methylcarbamyl, thiocarbamyl, methylthiocarbamyl, ##EQU10## HOCH₂ CH₂ CO--, HOOCCH₂ CO-- and H₂ NOC--CH₂ CO-- and the amino group of the B₂₉ amino acid (lysine) is either free or protected by an acyl or other blocking substituent containing no more than 4 carbon atoms and no free primary amino group selected from the group consisting of formyl, acetyl, trifluoroacetyl, cyclopropanecarbonyl, acetoacetyl, carbamyl, methylcarbamyl, thiocarbamyl, methylthiocarbamyl, ##EQU11## HOCH₂ CH₂ CO--, HOOC--CH₂ --CO-- and H₂ NOC--CH₂ CO--.

17. A method according to claim 16 in which the terminal amino group of the A chain (A₁ glycine) is either free or protected by an acyl or other blocking substituent containing no more than three atoms other than hydrogen selected from the group consisting of formyl, acetyl, carbamyl, thiocarbamyl, ##EQU12## and the amino group of the B₂₉ amino acid (lysine) is either free or protected by an acyl or other blocking substituent containing no more than three atoms other than hydrogen selected from the group consisting of formyl, acetyl, carbamyl, thiocarbamyl, ##EQU13##

18. A method according to claim 16 in which the B₁ amino acid is protected by acetyl or acetoacetyl.

19. A method according to claim 16 in which the B₁ amino acid is protected by 2,2-dimethyl-3-formyl-L-thiazolidine-4-carbonyl.

20. A method according to claim 16 in which at least one of the substituents is a carbamyl group.

21. A method according to claim 16 wherein the insulin is mono-substituted.

22. A method according to claim 16 wherein the insulin is di-substituted.

23. A method according to claim 16 wherein the insulin is tri-substituted.

24. A method according to claim 17 wherein the A₁ and B₁ protective substituents are the same.

25. A method according to claim 16 wherein the insulin is B₁ (phenylalanine-N-monocarbamyl insulin.

26. A method according to claim 16 wherein the insulin is A₁ (glycine)B₁ (phenylalanine)-N,N'"'"'-dicarbamyl insulin.

27. A method according to claim 16 wherein the insulin is A₁ (glycine)B₁ (phenylalanine)B₂₉ (lysine)-N,N'"'"',N"-tricarbamyl insulin.

29. A method of treating diabetes in humans which comprises parenterally administering to such human a hypoglycemically effective amount of a mixture of A₁ (glycine), B₁ (phenylalanine), B₂₉ (lysine)-N,N'"'"'N"-tricarbamyl insulin and A₁ (glycine), B₁ (phenylalanine)-N,N'"'"'-dicarbamyl insulin.