Claims
- 1. An aqueous insulin preparation comprising:
- (i) dissolved and/or precipitated protein selected from the group consisting of human insulin, an analogue of human insulin, a derivative of human insulin, and mixtures of the foregoing; and
- (ii) a water-soluble reduced or non-reducing carbohydrate containing at least 4 carbon atoms in the main carbohydrate structure, or a water-soluble non-reducing ester and/or ether derivative of a carbohydrate or reduced carbohydrate containing at least 4 carbon atoms in the main carbohydrate structure, or mixtures thereof, wherein said carbohydrate is present in an amount corresponding to 100 to 400 mM.
- 2. The insulin preparation of claim 1, wherein said carbohydrate or carbohydrate derivative contains from 5 to 18 carbon atoms in the main carbohydrate structure.
- 3. The insulin preparation of claim 2, wherein said carbohydrate is selected from the group consisting of mannitol, sorbitol, xylitol, trehalose, sucrose, and any mixture thereof.
- 4. The insulin preparation of claim 3, wherein said carbohydrate is mannitol.
- 5. The insulin preparation of claim 1, which further comprises a halogenide.
- 6. The insulin preparation of claim 5, wherein the halogenide is a chloride.
- 7. The insulin preparation of claim 6, wherein the chloride is sodium chloride.
- 8. An insulin preparation according to claim 7, wherein said sodium chloride is present at a concentration of 0 to 100 mM.
- 9. The insulin preparation of claim 1, wherein said analogue of human insulin is selected from the group consisting of: (i) human insulin in which position B28 is Asp, Lys, Leu, Val or Ala and position B29 is Lys or Pro; des (B28-B30) human insulin; des(B27) human insulin; and des(B30) human insulin.
- 10. The insulin preparation of claim 9, wherein said analogue of human insulin is an insulin in which position B28 is Asp or Lys, and position B29 is Lys or Pro.
- 11. The insulin preparation of claim 10, wherein the analogue is Asp.sup.B28 human insulin or Lys.sup.B28 Pro.sup.B29 human insulin.
- 12. The insulin preparation of claim 1, wherein said derivative of human insulin has one or more lipophilic subsitituents.
- 13. The insulin preparation of claim 12, wherein the derivative is an acylated insulin.
- 14. The insulin preparation of claim 1, wherein said derivative of human insulin is selected from the group consisting of:
- B29-N.sup..epsilon. -myristoyl-des(B30)-humaninsulin, B29-N.sup..epsilon. -myristoylhumaninsulin, B29-N.sup..epsilon. -palmitoyl human insulin, B28-N.sup..epsilon. -myristoyl Lys.sup.B28 Pro.sup.B29 human insulin, B28-N.sup..epsilon. -palmitoyl Lys.sup.B28 Pro.sup.B29 human insulin, B30-N.sup..epsilon. -myristoyl-Thr.sup.B29 Lys.sup.B30 -human insulin, B30-N.sup..epsilon. -palmitoyl-Thr.sup.B29 Lys.sup.B30 -human insulin, B29-N.sup..epsilon. -(N-palmitoyl-.gamma.-glutamyl)-des(B30-human insulin, B29-N.sup..epsilon. -(N-lithocholyl-.gamma.-glutamyl)-des(B30)-human insulin and B29-N.sup..epsilon. -(.omega.-carboxyheptadecanoyl)-des(B30)-human insulin.
- 15. The insulin preparation of claim 14, wherein the derivative is B29-N.sup..epsilon. -myristoyl-des(B30)-human insulin.
- 16. The insulin preparation of claim 1, wherein said precipitated protein comprises crystals comprising (i) insulin, insulin analogue or insulin derivative, and (ii) protamine.
- 17. The insulin preparation of claim 16, in which the crystals further comprise zinc and optionally one or more phenolic compounds.
- 18. The insulin preparation of claim 1, wherein said dissolved and precipitated insulin or insulin analogue is present in a weight ratio of 1:99 to 99:1.
- 19. The insulin preparation of claim 18, wherein the precipitated insulin or insulin analogue is crystalline.
- 20. The insulin preparation of claim 18, wherein said weight ratio is 20:80 to 80:20.
- 21. The insulin preparation of claim 20, wherein said weight ratio is 30:70 to 70:30.
- 22. The insulin preparation of claim 1, in which the carbohydrate is mannitol.
- 23. The insulin preparation of claim 22, wherein the carbohydrate is present in an amount corresponding to 150 to 250 mM.
- 24. The insulin preparation of claim 23, wherein the carbohydrate is present in an amount corresponding to 180 to 230 mM.
- 25. The insulin preparation of claim 1, wherein said human insulin or insulin analogue or derivative is present in an amount of 60 to 3000 nmol/ml.
- 26. The insulin preparation of claim 25, wherein the amount of human insulin or insulin analogue or derivative is 240 to 1200 nmol/ml.
- 27. The insulin preparation of claim 8, wherein said sodium chloride is present at a concentration of 5 to 40 mM.
- 28. The insulin preparation of claim 27, wherein said concentration is 5 to 20 mM.
- 29. The insulin preparation of claim 1, further comprising a physiologically tolerated buffer.
- 30. The insulin preparation of claim 29, wherein the buffer is a phosphate buffer.
- 31. A parenteral pharmaceutical formulation comprising the insulin preparation of claim 15.
- 32. A method for preparing an insulin preparation containing both dissolved and precipitated insulin analogue, comprising the steps of:
- a) providing an acidic solution comprising an analogue of human insulin, zinc, and a sub-isophanic amount of protamine;
- b) providing an alkaline solution comprising a substance which acts as a buffer at physiological pH;
- wherein the solution of step a) and/or step b) comprises a water-soluble reduced or non-reducing carbohydrate containing at least 4 carbon atoms in the main carbohydrate structure, or a water-soluble non-reducing ester and/or ether derivative of a carbohydrate or reduced carbohydrate containing at least 4 carbon atoms in the main carbohydrate structure, or mixtures thereof, in an amount corresponding to 100 to 400 mM and wherein at least one of the above solutions further comprises a phenolic compound;
- c) mixing the acidic and alkaline solutions and, optionally, adjusting the pH to a value in the range of 6.5 to 8.0; and
- d) leaving the resulting suspension for precipitation.
- 33. The method of claim 32, wherein the weight ratio of insulin analogue to protamine in the solution of step a) is selected so as to obtain in the final product a weight ratio of dissolved to precipitated insulin analogue in the range of 1:99 to 99:1.
- 34. The method of claim 33, wherein the solution of step a) comprises 120 to 6000 nmol/ml of insulin analogue and 0.01 to 5.0 mg/ml of protamine.
- 35. The method of claim 32, wherein said zinc is present in the solution of step a) in an amount corresponding; to 10-40 .mu.g zinc/100 IU insulin.
- 36. The method of claim 32, wherein solution a) and/or solution b) comprises chloride in an amount corresponding to 0 to 100 mM in the final product.
- 37. The method of claim 32, wherein the pH of the acidic solution of step a) is below 5.
- 38. The method of claim 32, wherein the insulin analogue is selected from the group consisting of: human insulin wherein position B28 is iasp, Lys, Leu, Val or Ala and position B29 is Lys or Pro; des (B28-B30) human insulin, des(B27) human insulin and des(B30) human insulin.
- 39. The method of claim 38, wherein the insulin analogue is selected from the group consisting of Asp.sup.B28 human insulin and Lys.sup.B28 Pro.sup.B29 human insulin.
- 40. The method of claim 32, wherein the phenolic compound is selected from the group consisting of phenol, m-cresol or a mixture thereof.
- 41. The method of claim 32, wherein said carbohydrate or carbohydrate derivative contains from 5 to 18 carbon atoms in the main carbohydrate structure.
- 42. The method of claim 41, wherein said carbohydrate or carbohydrate derivative is selected from the group consisting of mannitol, sorbitol, xylitol, trehalose, sucrose, and any mixture thereof.
- 43. The method of claim 32, wherein the buffer substance employed in the alkaline solution of step b) is a physiologically tolerated buffer.
- 44. The method of claim 32, wherein the precipitated insulin analogue is in the form of crystals comprising insulin analogue and protamine.
- 45. The method of claim 32, wherein the suspension obtained in step d) is left at a temperature in the range of 5.degree. C. to 40.degree. C. for precipitation.
- 46. The method of claim 32, wherein in step (c) the pH is adjusted to between 7.0 and 7.8.
- 47. The method of claim 33, wherein the insulin or insulin analogue is in the weight ratio of 20:80 to 80:20.
- 48. The method of claim 34, wherein the insulin or insulin analogue is in the weight ratio of 30:70 to 70:30.
- 49. The method of claim 35, wherein said zinc is zinc chloride.
- 50. The method of claim 35, wherein the amount of zinc corresponds to 15-35 .mu.g zinc/100 IU insulin.
- 51. The method of claim 36, wherein said chloride is sodium chloride.
- 52. The method of claim 51, wherein said chloride is present in an amount corresponding to 5 to 20 mM in the final product.
- 53. The method of claim 37, wherein the pH of the acidic solution of step a) is in the range of 2 to 3.5.
- 54. The method of claim 39, wherein the insulin analogue is Asp.sup.B28 human insulin.
- 55. The method of claim 42, wherein said carbohydrate or carbohydrate derivative is selected from the group consisting of mannitol, sorbitol, and mixtures thereof.
- 56. The method of claim 43, wherein the buffer is a phosphate buffer.
- 57. The method of claim 56, wherein the buffer is disodium phosphate dihydrate.
- 58. The method of claim 45, wherein the temperature is 20.degree. C. to 36.degree. C.
- 59. The method of claim 58, wherein the temperature is 30.degree. C. to 34.degree. C.
Priority Claims (2)
Number |
Date |
Country |
Kind |
684/96 |
Jun 1996 |
DKX |
|
899/96 |
Aug 1996 |
DKX |
|
CROSS-REFERENCE TO RELATED APPLICATIONS
This application claims priority under 35 U.S.C. .sctn. 119(e) to U.S. provisional application Ser. No. 60/023,264 filed Jun., 29, 1996 and U.S. provisional application Ser. No. 60/024,862 filed Aug. 28, 1996 and claims priority under 35 U.S.C. 119 of Danish application 684/96 filed Jun. 20, 1996 and 899/96 filed Aug. 27, 1996, the contents of which applications are fully incorporated herein by reference.
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Non-Patent Literature Citations (1)
Entry |
Gupta and Bhat "Effect of solvent additives on the thermal stability of insulin" Center for Biotechnology, New Delhi pp. 209-212. |