Preparation of stable zinc insulin analog crystals

• US 5,504,188 A
• Filed: 06/16/1994
• Issued: 04/02/1996
• Est. Priority Date: 06/16/1994
• Status: Expired due to Term

• Current Assignee: Eli Lilly and Company
• Original Assignee: Eli Lilly and Company
• Inventors: Carter, Nancy D., Baker, Jeffrey C., Frank, Bruce H.
• Primary Examiner(s): Warden, Jill
• Assistant Examiner(s): Prickril, Benet
• Application Number:
  US08/260,647
• Time in Patent Office: 656 Days
• Field of Search: 530/304, 514/3
• US Class Current: 530/304
• 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: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 5/00 Drugs for disorders of the endocrine system
• CPC Class Codes: A61P 5/48 of the pancreatic hormones
• CPC Class Codes: A61P 5/50 for increasing or potentiating the activity of insulin
• CPC Class Codes: C07K 14/62 Insulins

Abstract

The present invention discloses a process of preparing a crystalline insulin analog. The process is useful in the purification and manufacture of Lys^B28 Pro^B29 -human insulin. Lys^B28 Pro^B29 -human insulin is useful in the treatment of diabetes.

Claims

1. A process of preparing crystalline Lys^B28 Pro^B29 -human insulin, which comprises:

crystallizing Lys^B28 Pro^B29 -human insulin from a solution comprising Lys^B28 Pro^B29 -human insulin, zinc, at least 0.3N of an organic acid selected from the group consisting acetic, citric, or glycine, and a phenolic at a pH of about 5.5 to about 6.5.

2. The process of claim 1, wherein the concentration of Lys^B28 Pro^B29 -human insulin is about 1.8 to about 2.5 g/l.

3. The process of claim 2, wherein the phenolic is phenol.

4. The process of claim 3, wherein the concentration of zinc is about 40 mg/gm of Lys^B28 Pro^B29 -human insulin to about 400 mg/gm of Lys^B28 Pro^B29 -human insulin.

5. The process of claim 4, wherein the concentration of phenol is about 0.15% to 0.25% on a volume basis.

6. The process of claim 5, wherein the pH is about 5.9 to about 6.2.

7. The process of claim 6, wherein the Lys^B28 Pro^B29 -human insulin is recrystallized.

8. The process of claim 1, wherein the crystallization is initiated by adjusting the basicity of the solution comprising Lys^B28 Pro^B29 -human insulin, zinc, at least 0.3N acetic acid, and a phenolic;

to a pH of about 5.9 to about 6.2 with a physiologically tolerated base.

9. The process of claim 1, wherein the crystallization is initiated by the addition of zinc to the solution at pH 5.9 to 6.2.

10. A process of preparing crystalline Lys^B28 Pro^B29 -human insulin, which comprises:

(a) adding phenol at 0.15% to 0.25% on a final volume basis to a solution of Lys^B28 Pro^B29 -hI solution at about 1.8 to 2.5 g/L in about 1N acetic acid;

(b) adjusting the pH to about 5.9 to about 6.2 with ammonium hydroxide; and

(c) adding a zinc salt solution to a final concentration of about 40 to about 400 mg zinc chloride per gram of Lys^B28 Pro^B29 -hI.

11. The process of claim 10, wherein the zinc salt is zinc chloride.