GLP-2 formulations
View Patent ↗The invention is directed to formulations of GLP-2 peptides and analogs thereof exhibiting superior stability following storage and/or exposure to elevated temperatures. The GLP-2 compositions comprise a GLP-2 peptide or an analog thereof, a phosphate buffer, L-histidine, and mannitol.
1. A glucagon-like peptide 2 (GLP-2) formulation comprising:
(a) a medically useful amount of a naturally occurring GLP-2 or an analog thereof;
(b) a phosphate buffer in an amount sufficient to adjust the pH of the formulation to a physiologically tolerable level;
(c) L-histidine; and
(d) a bulking agent selected from the group consisting of mannitol and sucrose.
2. The GLP-2 formulation of claim 1 , wherein the pH of the formulation is greater than about 6.0.
3. The GLP-2 formulation of claim 2 , wherein the GLP-2 peptide is h(Gly2)GLP-2.
4. The GLP-2 formulation according of claim 2 , wherein the pH of the formulation is from about 6.9 to about 7.9.
5. The GLP-2 formulation of claim 4 , wherein the GLP-2 peptide is h(Gly2)GLP-2.
6. The GLP-2 formulation of claim 4 , wherein the pH of the formulation is from about 7.3 to about 7.4.
7. The GLP-2 formulation of claim 6 , wherein the GLP-2 peptide is h(Gly2)GLP-2.
8. The GLP-2 formulation of claim 1 , wherein the GLP-2 peptide or analog thereof is present at a concentration of about 0.1 to about 50 mg/ml.
9. The GLP-2 formulation of claim 8 , wherein the GLP-2 peptide is h(Gly2)GLP-2.
10. The GLP-2 formulation of claim 8 , wherein the GLP-2 peptide or analog thereof is present at a concentration of about 5 to about 40 mg/ml.
11. The GLP-2 formulation of claim 10 , wherein the GLP-2 peptide is h(Gly2)GLP-2.
12. The GLP-2 formulation of claim 10 , wherein the GLP-2 peptide or analog thereof is present at a concentration of about 7 to about 30 mg/ml.
13. The (GLP-2 formulation of claim 12 , wherein the GLP-2 peptide is h(Gly2)GLP-2.
14. The GLP-2 formulation of claim 12 , wherein the GLP-2 peptide or analog thereof is present at a concentration of about 10 to about 20 mg/ml.
15. The GLP-2 formulation of claim 14 , wherein the GLP-2 peptide is h(Gly2)GLP-2.
16. The GLP-2 formulation of claim 14 , wherein the L-histidine is present in an amount of about 0.5 to about 1%.
17. The GLP-2 formulation of claim 16 , wherein the GLP-2 peptide is h(Gly2)GLP-2.
18. The GLP-2 formulation of claim 16 , wherein the bulking agent is mannitol.
19. The GLP-2 formulation of claim 18 , wherein the GLP-2 peptide is h(Gly2)GLP-2.
20. The GLP-2 formulation of claim 18 , wherein the mannitol is present at a concentration of about 2 to about 5%.
21. The GLP-2 formulation of claim 20 , wherein the GLP-2 peptide is h(Gly2)GLP-2.
22. The GLP-2 formulation of claim 20 , wherein the mannitol is present at a concentration of about 2.5 to about 3.5%.
23. The GLP-2 formulation of claim 22 , wherein the GLP-2 peptide is h(Gly2)GLP-2.
24. The GLP-2 formulation of claim 1 , wherein the GLP-2 peptide is selected from the group consisting of a mammalian GLP-2 peptide, a vertebrate GLP-2 peptide, and a human GLP-2 peptide.
25. The GLP-2 formulation of claim 24 , wherein the GLP-2 peptide is h(Gly2)GLP-2.
26. The GLP-2 formulation of claim 24 , wherein the GLP-2 peptide has the sequence of a GLP-2 species from an animal selected from the group consisting of a primate, rat, mouse, porcine species, oxine species, bovine species, degu, hamster, guinea pig, fish, chicken, and human.
27. The GLP-2 formulation of claim 26 , wherein the GLP-2 peptide is h(Gly2)GLP-2.
28. The GLP-2 formulation of claim 27 , which is stable at ambient temperature for up to 6 months, as evidenced by GLP-2 peptide degradation of less than about 5% during this time period.
29. The GLP-2 formulation of claim 28 , wherein less than about 4% peptide degradation is observed after storage of the GLP-2 formulation during the time period.
30. The GLP-2 formulation of claim 29 , wherein less than about 2% peptide degradation is observed after storage of the GLP-2 formulation during the time period.
31. The GLP-2 formulation of claim 1 , wherein the GLP-2 analog is identified by a process comprising:
(a) screening peptides against cells genetically engineered to produce the GLP-2 receptor, and
(b) identifying peptides which bind to the GLP-2 receptor, wherein such peptides are identified as GLP-2 peptides useful in the formulation of claim 1 .
32. The GLP-2 formulation of claim 1 , wherein the GLP-2 peptide is an analog of natural GLP-2, the analog having:
(a) one or more amino acid substitutions, additions, deletions, or modifications; and
(b) GLP-2 receptor binding activity.
33. The GLP-2 formulation of claim 1 , wherein the GLP-2 peptide is an analog which has been altered to confer resistance to endogenous enzymes.
34. The GLP-2 formulation of claim 33 , wherein the alteration comprises substitution of the alanine residue at position 2 of GLP-2 with another suitable amino acid.
35. The GLP-2 formulation of claim 34 , wherein the alanine residue at position 2 is substituted with glycine or serine.
36. The GLP-2 formulation of claim 1 , wherein the GLP-2 analog is a GLP-2 receptor antagonist.
37. The GLP-2 formulation of claim 36 , wherein the GLP-2 receptor antagonist has either (1) an amino acid substitution selected from the group consisting of Asp 15 , Phe 22 , Thr 29 , Thr 32 , Asp 33 , and combinations thereof; or (2) an amino acid substitution of Ala at position 2 by an amino acid selected from the group consisting of Leu, Cys, Glu, Arg, Trp and PO 3 —-Tyr, wherein the residue or position for the amino acid substitution is numbered according to a mammalian GLP-2.
38. The GLP-2 formulation of claim 1 in lyophilized form.
39. The lyophilized formulations of claim 38 , comprising less than about 5% water by weight.
40. The lyophilized formulations of claim 39 , comprising 2% or less water by weight.
41. The GLP-2 formulation of claim 1 , which is stable at a temperature of about 4° C. for up to 18 months, as evidenced by GLP-2 peptide degradation of less than about 5% during this time period.
42. The GLP-2 formulation of claim 41 , wherein less than about 4% peptide degradation is observed after storage of the GLP-2 during the time period.
43. The GLP-2 formulation of claim 42 , wherein less than about 2% peptide degradation is observed after storage of the GLP-2 formulation during the time period.
44. The GLP-2 formulation of claim 1 , wherein said GLP-2 analog has one or more amino acid substitutions, additions, deletions, or modifications and has GLP-2 receptor binding activity.
45. The GLP-2 formulation of claim 1 , wherein the GLP-2 peptide is h(Gly2)GLP-2.
46. A GLP-2 formulation comprising:
(a) about 0.1 to about 50 mg/ml of a GLP-2 peptide or an analog thereof;
(b) a phosphate buffer in an amount sufficient to adjust the pH of the formulation to a pharmaceutically tolerable level;
(c) about 0.5 to about 1% L-histidine; and
(d) about 2 to about 5% mannitol.
47. The GLP-2 formulation of claim 46 , wherein the GLP-2 is h(Gly2)GLP-2.
48. The GLP-2 formulation of claim 47 , wherein the formulation is lyophilized.
49. The GLP-2 formulation of claim 47 , wherein the pH of the formulation is selected from the group consisting of greater then about 6.0, and from about 6.9 to about 7.9.
50. The GLP-2 formulation of claim 49 , wherein the pH of the formulation is from about 7.3 to about 7.4.
51. The GLP-2 formulation of claim 46 , wherein said GLP-2 analog has one or more amino acid substitutions, additions, deletions, or modifications and has GLP-2 receptor binding activity.
52. A GLP-2 formulation comprising:
(a) a medically useful amount of a naturally occurring GLP-2 peptide or an analog thereof;
(b) a phosphate buffer in an amount sufficient to adjust the pH of the formulation to a physiologically tolerable level;
(c) L-histidine in an amount sufficient to stabilize the formulation; and
(d) a bulking agent selected from the group consisting of mannitol and sucrose.
53. A method for making a lyophilized formulation of GLP-2 comprising the following steps:
(a) preparing a GLP-2 formulation comprising:
(i) a GLP-2 peptide or an analog thereof;
(ii) a phosphate buffer in an amount sufficient to adjust the pH of the formulation to a pharmaceutically tolerable level;
(iii) L-histidine; and
(iv) a hulking agent selected from the group consisting of mannitol and sucrose;
(b) freezing the formulation to −40° C.;
(c) drying the formulation in a first drying step at −20° C; and
(d) drying the formulation in a second drying step at +20° C.
54. The method of claim 53 , wherein the pH of the GLP-2 formulation prior to freezing is selected from the group consisting of greater than about 6.0, and from about 6.9 to about 7.9.
55. The method of claim 54 , wherein the pH of the formulation is from about 7.3 to about 7.4.
56. The method of claim 53 , wherein the freezing process of step (b) comprises:
(a) cooling the formulation from ambient temperature to about −1° C. at about 2° C./minute, followed by maintaining the formulation at about −1° C. for about 15 minutes; and
(b) cooling the formulation from about −1° C. to about −40° C. at about 2° C./minute, followed by maintaining the formulation at about −40° C. for about 4 hours.
57. The method of claim 53 , wherein the drying process of step (c) comprises:
(a) raising the temperature from about −40° C. to about −20° C. at about 2° C./minute; and
(b) maintaining the formulation at about −20° C. for about 14 hours under a vacuum of about 150 mT with a condenser temperature of about −80° C.
58. The method of claim 53 , wherein the drying process of step (d) comprises:
(a) warming the formulation from about −20° C. to about +20° C. at about 2° C./minute;
(b) maintaining the formulation at about +20° C. for about 14 hours at a vacuum of about 150 mT and a condenser temperature of about −80° C. until there is less than about 5% of water remaining in the formulation.
59. The method of claim 58 , wherein the formulation is maintained at about +20° C., at a vacuum of about 150 mT and a condenser temperature of about −80° C., until there is about 2% or less of water remaining in the formulation.
60. The method of claim 53 , wherein said GLP-2 analog has one or more amino acid substitutions, additions, deletions, or modifications and has GLP-2 receptor binding activity.
61. A kit comprising:
(a) a lyophilized GLP-2 formulation comprising:
(i) a GLP-2 peptide or an analog thereof;
(ii) a phosphate buffer in an amount sufficient to adjust the pH of the formulation to a pharmaceutically acceptable level;
(iii) L-histidine; and
(iv) a bulking agent selected from the group consisting of mannitol and sucrose;
(b) a vial of sterile water for reconstitution; and
(c) instructions directing reconstitution.
62. The kit of claim 61 , wherein the pH of the GLP-2 formulation is selected from the group consisting of greater than about 5.5, greater than about 6.0, and from about 6.9 to about 7.9.
63. The kit of claim 62 , wherein the pH of the formulation is from about 7.3 to about 7.4.
64. The kit of claim 63 , wherein the GLP-2 peptide is h(Gly2)GLP-2.
65. A kit of claim 61 , further comprising an injection device for administration.
66. The kit of claim 61 , wherein following reconstitution the GLP-2 formulation is stable for at least about 12 hours.
67. The kit of claim 61 , wherein following reconstitution the GLP-2 formulation is stable for up to about 24 hours.
68. The kit of claim 61 , wherein said GLP-2 analog has one or more amino acid substitutions, additions, deletions, or modifications, and has GLP-2 receptor binding activity.
69. A method for treating a human or animal having a gastrointestinal disorder, disease or condition for which treatment with GLP-2 is indicated, the method comprising the step of administering a therapeutically effective amount of a GLP-2 formulation comprising:
(a) a GLP-2 peptide or an analog thereof;
(b) a phosphate buffer in an amount sufficient to adjust the pH of the formulation to a pharmaceutically tolerable level;
(c) L-histidine; and
(d) a bulking agent selected from the group consisting of mannitol and sucrose,
thereby enhancing, maintaining, or promoting the growth or functioning of the gastrointestinal tract.
70. The method of claim 69 , wherein the pH of the GLP-2 formulation is selected from the group consisting of greater than about 5.5, greater than about 6.0, and from about 6.9 to about 7.9.
71. The method of claim 70 , wherein the pH of the formulation is from about 7.3 to about 7.4.
72. The method of claim 70 , wherein the GLP-2 peptide is h(Gly2)GLP-2.
73. The method of claim 69 , wherein the GLP-2 formulation is administered by injection.
74. The method of claim 69 , wherein the GLP-2 formulation is administered by infusion.
75. The method of claim 69 , wherein said GLP-2 analog has one or more amino acid substitutions, additions, deletions, or modifications and has GLP-2 receptor binding activity.