Mono-, di, and N.sub.A1, N.sup.B1, N.sup.B29 -tri-acylated insulin

• US 3,869,437 A
• Filed: 08/21/1973
• Issued: 03/04/1975
• Est. Priority Date: 05/08/1970
• Status: Expired due to Term

• Current Assignee: David Gordon Lindsay, Sydney Shall
• Original Assignee: David Gordon Lindsay, Sydney Shall
• Inventors: Shall, Sydney, Lindsay, David Gordon
• Primary Examiner(s): Gotts, Lewis
• Assistant Examiner(s): Suyat, Reginald J.
• Application Number:
  US05/390,216
• Time in Patent Office: 560 Days
• Field of Search: 260/112.7 424/178
• US Class Current: 530/303
• 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 seven 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 PHYSIOLOGICALLY-ACCEPTABLE, SUSTANTIALLY-PURE 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 SEVEN CARBON ATOMS SELECTED FROM THE GROUP CONSISTING OF FORMYL, ACETY, TRIFLUOROACETYL, CYCLOPROPANECARBONYL, ACETO-ACETYL, BENZOYL, 2,2-DIMETHYL-3-FORMYL-L-THIAZOLIDINE-4-CARBONYL, CARBAMYL, METHYLCARBAMYL, THIOCARBAMYL, METHYLTHIOCARBAMYL,

2. A physiologically-acceptable, substantially-pure mono-, di-or tri-substituted insulin 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,

3. An insulin according to claim 1, in which the B1 amino acid is protected by acetyl or acetoacetyl.

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

5. An insulin according to claim 1, in which at least one of the substituents is a carbamyl group.

6. An insulin according to claim 1 which is mono-substituted.

7. An insulin according to claim 1 which is di-substituted.

8. An insulin according to claim 7 in which A1 and B1 protective substituents are the same.

9. An insulin according to claim 1, which is tri-substituted.

10. An insulin according to claim 9 in which the A1, B1 and B29 protective substitutents are the same.

11. The insulin according to claim 1 which is A1, B1-diacetyl insulin.

12. Physiologically-acceptable, substantially-pure B1(phenylalanine)-N-monocarbamyl insulin.

13. Physiologically-acceptable, substantially-pure A1(glycine), B1(phenylalanine)-N,N'"'"''"'"'-dicarbamyl insulin.

14. Physiologically-acceptable, substantially-pure A1(glycine)B1(phenylalanine)B29(lysine)-N,N'"'"''"'"',N'"'"''"'"''"'"''"'"'-tricarbamyl insulin.

15. A process for producing a compound according to claim 1 which comprises reacting an insulin with an acylating agent or other blocking reagent to acylate or otherwise block the desired amino group with the desired substituent and in which when a mono-substituted insulin is desired from 1 to 2 moles of acylating agent or blocking reagent is used and when di- or tri-substitution is desired from 3 to 4 moles of acylating agent or blocking reagent is used per mole of insulin, and separating said insulin from the reaction mixture.

16. A process according to claim 15, in which the pH of the reaction medium is maintained at from 7 to 8.5 or 9.

17. A process according to claim 15 in which the pH of the reaction medium is maintained at from 7 to 8.5.