Process for the preparation and purification of peptides

• US 4,623,716 A
• Filed: 11/01/1984
• Issued: 11/18/1986
• Est. Priority Date: 11/01/1984
• Status: Expired due to Fees

• Current Assignee: USV Pharmaceutical Corporation
• Original Assignee: USV Pharmaceutical Corporation
• Inventors: Stevenson, David, Islam, Mohammad A.
• Primary Examiner(s): Phillips, Delbert R.
• Application Number:
  US06/667,025
• Time in Patent Office: 747 Days
• Field of Search: 260/112.5 T, 530/333, 530/307, 530/311
• US Class Current: 530/333
• 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 1/02 in solution C07K1/003, C07K1/006 take precedence
• CPC Class Codes: C07K 1/067 for sulfur-containing functions
• CPC Class Codes: C07K 14/585 Calcitonins
• CPC Class Codes: Y10S 930/15 Oxytocin or vasopressin; related peptides
• CPC Class Codes: Y10S 930/16 Somatostatin; related peptides

Abstract

Peptides, such as calcitonin, having two cysteine residues connected by a disulfide bond are prepared by a process which includes the step of cyclizing the cysteines at a pH of 8.5 to 9.0, and a concentration of 0.4 to 1.8 mg of peptide per ml of solution. These novel conditions produce rapid cyclization with an unforeseen improvement in yield.

Claims

1. A process for forming a disulfide bridge in a peptide containing at least one pair of cysteine moieties which are initially uncyclized, comprising maintaining a solution of the peptide at a pH above 8.5 but not exceeding 9.0, until the cyclization has taken place.

2. A process for forming a disulfide bridge in a peptide containing at least one pair of cysteine moieties which are initially uncyclized,

wherein

one of the cysteine moieties of the initially uncyclized peptide is a residue substituted with an alkyl group containing up to 6 carton atoms,

the concentration of the initially uncyclized peptide is 0.7 to 1.0 mg of peptide per ml of solution, and

the solution of peptide is maintained at a ph above 8.5 but not exceeding 9.0, until the cyclization has taken place.

3. The process of claim 2 wherein the peptide is a calcitonin having the structure of a naturally occurring calcitonin, or a deletion, substitution, or addition analog thereof.

4. the process of claim 2 wherein the calcitonin is human calcitonin or an analog thereof.