Methods of synthesizing insulin polypeptide-oligomer conjugates, and proinsulin polypeptide-oligomer conjugates and methods of synthesizing same

• US 20050080001A1
• Filed: 09/30/2004
• Published: 04/14/2005
• Est. Priority Date: 09/07/2001
• Status: Active Grant

• Current Assignee: Biocon Limited (Biocon Incorporated)
• Original Assignee: Biocon Limited (Biocon Incorporated)
• Inventors: Radhakrishnan, Balasingam, Ekwuribe, Nnochiri N., Soltero, Richard
• Granted Patent: US 7,611,864 B2
• Time in Patent Office: Days
• US Class Current: 435/68.100
• CPC Class Codes: A61K 47/60 the organic macromolecular compound being a polyoxyalkylene oligomer, polymer or dendrimer, e.g. PEG, PPG, PEO or polyglycerol
• CPC Class Codes: C07K 14/62 Insulins

Abstract

Methods for synthesizing proinsulin polypeptides are described that include a contacting a proinsulin polypeptide including an insulin polypeptide coupled to one or more peptides by peptide bond(s) capable of being cleaved to yield the insulin polypeptide with an oligomer under conditions sufficient to couple the oligomer to the insulin polypeptide portion of the proinsulin polypeptide and provide a proinsulin polypeptide-oligomer conjugate, and cleaving the one or more peptides from the proinsulin polypeptide-oligomer conjugate to provide the insulin polypeptide-oligomer conjugate. Methods of synthesizing proinsulin polypeptide-oligomer conjugates are also described as are proinsulin polypeptide-oligomer conjugates. Methods of synthesizing C-peptide polypeptide-oligomer conjugates are also described.

Claims

1-445. -445. (canceled)

446. A proinsulin polypeptide-oligomer conjugate comprising:

a proinsulin polypeptide comprising an insulin polypeptide; and

an oligomer comprising a hydrophilic moiety and a lipophilic moiety coupled to the insulin polypeptide portion of the proinsulin polypeptide.

447. The conjugate according to claim 446, wherein the proinsulin polypeptide comprises an insulin polypeptide having an A-chain polypeptide and a B-chain polypeptide;

a connecting peptide coupled at a first end to the C-terminus of the B-chain polypeptide and coupled at a second end to the N-terminus of the A-chain polypeptide; and

a leader peptide coupled to the N-terminus of the B-chain polypeptide.

448. The conjugate according to claim 446, wherein the connecting peptide is C-peptide.

449. The conjugate according to claim 446, wherein the connecting peptide is devoid of lysine residues.

450. The conjugate according to claim 446, wherein the proinsulin polypeptide is proinsulin.

451. The conjugate according to claim 446, wherein the insulin polypeptide is insulin.

452. The conjugate according to claim 446, wherein the oligomer is coupled to the insulin polypeptide portion of the proinsulin polypeptide at a lysine residue of the insulin polypeptide.

453. The conjugate according to claim 452, wherein the insulin polypeptide is insulin and wherein the lysine residue is at the B29 position of the insulin.

454. The conjugate according to claim 446, wherein the hydrophilic moiety is a polyalkylene glycol moiety.

455. The conjugate according to claim 454, wherein the polyalkylene glycol moiety is a polyethylene glycol moiety.

456. The conjugate according to claim 454, wherein the polyalkylene glycol moiety has between 4 and 10 polyalkylene glycol subunits.

457. The conjugate according to claim 446, wherein the lipophilic moiety is an alkyl or fatty acid moiety.

458. The conjugate according to claim 457, wherein the lipophilic moiety has between 3 and 8 carbon atoms.

459-467. -467. (canceled)