Details for Patent: 6,869,930

Title:	Acylated insulin
Abstract:	The present invention relates to protracted human insulin derivatives in which the A21 and the B3 amino acid residues are, independently, any amino acid residue which can be coded for by the genetic code except Lys, Arg and Cys; Phe.sup.B1 may be deleted; the B30 amino acid residue is (a) a non-codable, lipophilic amino acid having from 10 to 24 carbon atoms, in which case an acyl group of a carboxylic acid with up to 5 carbon atoms is bound to the .epsilon.-amino group of Lys.sup.B29 ; or (b) the B30 amino acid residue is deleted or is any amino acid residue which can be coded for by the genetic code except Lys, Arg and Cys, in any of which cases the .epsilon.-amino group of Lys.sup.B29 has a lipophilic substituent; and any Zn.sup.2+ complexes thereof with the proviso that when B30 is Thr or Ala and A21 and B3 are both Asn, and Phe.sup.B1 is present, then the insulin derivative is always present as a Zn.sup.2+ complex.
Inventor(s):	Havelund; Svend (Bagsvaerd, DK), Halstrom; John (Hundested, DK), Jonassen; Ib (Valby, DK), Andersen; Asser Sloth (Frederiksberg, DK), Markussen; Jan (Herlev, DK)
Assignee:	Novo Nordisk A/S (Bagsvaerd, DK)
Filing Date:	Sep 17, 1999
Application Number:	09/398,365
Claims:	1. An insulin derivative having the following sequence: ##STR2## wherein a) Xaa at positions A21 and B3 are, independently, any amino acid residue which can be coded for by the genetic code except Lys, Arg and Cys; b) Xaa at position B1 is Phe or is deleted; c) Xaa at position B30 is deleted; and d) the .epsilon.-amino group of Lys.sup.B29 is substituted with a lipophilic substituent having at least 6 carbon atoms. 2. The insulin derivative of claim 1, wherein Xaa at position A21 is Ala, Asn, Gln, Gly or Ser. 3. The insulin derivative of claim 2, wherein the lipophilic substituent has from 12 to 24 carbon atoms. 4. The insulin derivative of claim 1, wherein Xaa at position B1 is deleted. 5. The insulin derivative of claim 4, wherein the lipophilic substituent has from 12 to 24 carbon atoms. 6. The insulin derivative of claim 1, wherein Xaa at position B1 is Phe. 7. The insulin derivative of claim 6, wherein the lipophilic substituent has from 12 to 24 carbon atoms. 8. The insulin derivative of claim 1, wherein Xaa at position B3 is Asn, Asp, Gln or Thr. 9. The insulin derivative of claim 8, wherein the lipophilic substituent has from 12 to 24 carbon atoms. 10. The insulin derivative of claim 1, wherein Xaa at position A21 is Ala, Asn, Gln, Gly or Ser, and Xaa at position B3 is Asn, Asp, Gln or Thr. 11. The insulin derivative of claim 10, wherein the lipophilic substituent has from 12 to 24 carbon atoms. 12. The insulin derivative of claim 1, wherein Xaa at position A21 is Asn, Xaa at position B1 is Phe, and Xaa at position B3 is Asn. 13. The insulin derivative of claim 12, wherein the lipophilic substituent has from 12 to 24 carbon atoms. 14. The insulin derivative of claim 1, wherein the lipophilic substituent has from 12 to 24 carbon atoms. 15. The insulin derivative of claim 1, wherein the lipophilic substituent is cyclohexylvaleroyl. 16. The insulin derivative of claim 1, wherein the lipophilic substituent is acyl-glutamyl wherein the acyl is a linear, saturated acyl having 6 to 24 carbon atoms. 17. The insulin derivative of claim 1, wherein the lipophilic substituent is lauroyl. 18. The insulin derivative of claim 1, wherein the lipophilic substituent is myristoyl. 19. The insulin derivative of claim 1, wherein the lipophilic substituent is palmitoyl. 20. The insulin derivative of claim 1, wherein the lipophilic substituent is 2-succinylamido myristic acid. 21. The insulin derivative of claim 1, wherein the lipophilic substituent is 2-succinylamido palmitic acid. 22. The insulin derivative of claim 1, wherein the lipophilic substituent is 2-succinylamidoethyloxy palmitic acid. 23. The insulin derivative of claim 1, wherein the lipophilic substituent is myristoyl-.alpha.-glutamyl. 24. The insulin derivative of claim 1, wherein the lipophilic substituent is myristoyl-.alpha.-glutamyl-glycyl. 25. The insulin derivative of claim 1, wherein the lipophilic substituent is choloyl. 26. The insulin derivative of claim 1, wherein the lipophilic substituent is 7-deoxycholoyl. 27. The insulin derivative of claim 1, wherein the lipophilic substituent is lithocholoyl. 28. The insulin derivative of claim 1, wherein the lipophilic substituent is lithocholoyl-glutamyl. 29. The insulin derivative of claim 1, wherein the lipophilic substituent is 4-benzoyl-phenylalanine. 30. The insulin derivative of claim 1, wherein the lipophilic substituent is L-thyroxyl. 31. The insulin derivative of claim 1, wherein the lipophilic substituent is suberoyl-D-thyroxine. 32. The insulin derivative of claim 1, wherein the lipophilic substituent is 3,3',5,5'-tetraiodothyroacetyl. 33. The insulin derivative of claim 1, wherein the lipophilic substituent is an acyl group having at least 10 carbon atoms. 34. The insulin derivative of claim 33, wherein the lipophlic substituent is tetradecanoyl or hexadecanoyl. 35. The insulin derivative of claim 1, which is in the form of a hexamer. 36. The insulin derivative of claim 35, wherein the lipophilic substituent has from 12 to 24 carbon atoms. 37. The insulin derivative of claim 35, wherein Xaa at position A21 is Asn, Xaa at position B3 is Asn, and Xaa at position B1 is Phe. 38. The insulin derivative of claim 35, wherein two zinc ions bind to the hexamer. 39. The insulin derivative of claim 38, wherein the lipophilic substituent has from 12 to 24 carbon atoms. 40. The insulin derivative of claim 35, wherein three zinc ions bind to the hexamer. 41. The insulin derivative of claim 40, wherein the lipophilic substituent has from 12 to 24 carbon atoms. 42. The insulin derivative of claim 35, wherein four zinc ions bind to the hexamer. 43. The insulin derivative of claim 42, wherein the lipophilic substituent has from 12 to 24 carbon atoms. 44. The insulin derivative of claim 12, wherein the lipophilic substituent is tetradecanoyl. 45. A pharmaceutical composition which is an aqueous solution, said composition comprising (a) the insulin derivative of claim 44, (b) an isotonic agent, (c) a preservative and (d) a buffer. 46. The pharmaceutical composition of claim 45, wherein the pH of the aqueous solution is in the range of 6.5-8.5. 47. The pharmaceutical composition of claim 45, wherein the solubility of the insulin derivative exceeds 600 nmol/ml of the aqueous solution. 48. The pharmaceutical composition of claim 45, said composition further comprising an insulin or an insulin analogue which has a rapid onset of action. 49. The pharmaceutical composition of claim 45, wherein the insulin derivative is in the form of a hexamer. 50. A method of treating diabetes in a patient in need of such a treatment, said method comprising administering to the patient a therapeutically effective amount of a pharmaceutical composition of claim 45. 51. A method of treating diabetes in a patient in need of such a treatment, said method comprising administering to the patient a therapeutically effective amount of a pharmaceutical composition of claim 48. 52. A pharmaceutical composition which is an aqueous solution, said composition comprising (a) an insulin derivative of claim 1, (b) an isotonic agent, (c) a preservative and (d) a buffer. 53. The pharmaceutical composition of claim 52, wherein the pH of the aqueous solution is in the range of 6.5-8.5. 54. The pharmaceutical composition of claim 52, wherein the solubility of the insulin derivative exceeds 600 nmol/ml of the aqueous solution. 55. The pharmaceutical composition of claim 52, said composition further comprising an insulin or an insulin analogue which has a rapid onset of action. 56. The pharmaceutical composition of claim 52, wherein Xaa at position A21 is Asn, Xaa at position B3 is Asn, and Xaa at position B1 is Phe. 57. The pharmaceutical composition of claim 52, wherein the lipophilic substituent has from 12 to 24 carbon atoms. 58. The pharmaceutical composition of claim 52, wherein the insulin derivative is in the form of a hexamer. 59. A method of treating diabetes in a patient in need of such a treatment, said method comprising administering to the patient a therapeutically effective amount of a pharmaceutical composition of claim 52. 60. A method of treating diabetes in a patient in need of such a treatment, said method comprising administering to the patient a therapeutically effective amount of a pharmaceutical composition of claim 55.