Preparation of a polymer/interleukin-2 conjugate

• US 5,089,261 A
• Filed: 09/29/1989
• Issued: 02/18/1992
• Est. Priority Date: 01/23/1989
• Status: Expired due to Term

• Current Assignee: Chiron Corporation (Bayer AG)
• Original Assignee: Cetus Corporation (Bayer AG)
• Inventors: Aldwin, Lois, Katre, Nandini, Nitecki, Danute E., Goodson, Robert J.
• Primary Examiner(s): Schain, Howard E.
• Assistant Examiner(s): PERKINS, SUSAN
• Application Number:
  US07/415,046
• Time in Patent Office: 872 Days
• Field of Search: 424/85.1, 424/78, 424/83, 424/85.2, 530/351, 530/402, 530/409, 514/2
• US Class Current: 424/85.2
• CPC Class Codes: A61K 47/60 the organic macromolecular compound being a polyoxyalkylene oligomer, polymer or dendrimer, e.g. PEG, PPG, PEO or polyglycerol

Abstract

The present invention is a process for preparing a pharmaceutical composition comprising a biologically active conjugated protein. It comprises a polyethylene glycol or a polyoxyethylated polyol conjugated to IL-2. This protein is conjugated to reduce its immunogenicity, and increase its solubility, and increase its circulating in vivo half-life.

Claims

1. A pharmaceutically and therapeutically useful protein comprising a PEG or a polyoxyethylated polyol IL-2 conjugate made by contacting a PEG or a polyoxyethylated polyol, which has at least one hydroxyl group, with a chloroformate under the appropriate reaction conditions to form a PEG or a polyoxyethylated polyol active ester;

and contacting the PEG or the polyoxyethylated polyol active ester with IL-2 under the appropriate reaction conditions to form a PEG/IL-2 conjugate or a polyoxyethylated polyol/IL-2 conjugate.

2. A PEG or a polyoxyethylated polyol IL-2 conjugate comprising PEG or a polyoxyethylated polyol covalently conjugated to IL-2 through a urethane bond.

4. A conjugate in accordance with claim 2, wherein the conjugate has a molar ratio of from 1 and 3 PEGs per one IL-2.

5. A conjugate in accordance with claim 2, wherein the conjugate has a molar ratio of from 2 and 3 PEGs per one IL-2.

6. A conjugate in accordance with claim 2, wherein the IL-2 is a mutein.

7. A conjugate in accordance with claim 6, wherein the IL-2 mutein has up to six N-terminal deletions, and the cystein at position 125 is replaced by serine.

3. A PEG or a polyoxyethylated polyol IL-2 conjugate having the structure:

##STR4## wherein R is the PEG or the polyoxyethylated polyol.

8. A therapeutically useful formulation comprising a PEG/IL-2 conjugate, wherein the PEG is covalently attached to IL-2 via a urethane bond, and at least one pharmaceutically acceptable excipient.

10. A therapeutically useful formulation in accordance with claim 8, wherein the at least one excipient comprises amino acids, polymers, polyols, sugars, buffers, preservatives, or other proteins.

11. A therapeutically useful formulation in accordance with claim 10, wherein the excipients are selected from the group consisting essentially of octylphenoxy polyethoxy ethanol compounds, polyethylene glycol monostereate compounds, polyoxyethylene sorbitan fatty acid esters, sucrose, fructose, dextrose, maltose, glucose, dextran, mannitol, sorbitol, inositol, galactitol, xylitol, lactose, trehalose, bovine or human serum albumin, citrate, acetate, Ringer'"'"'s and Hank'"'"'s solutions, saline, phosphate, cysteine, arginine, carnitine, alanine, glycine, lysine, valine, leucine, polyvinylpyrrilolidone, or polyethylene glycol.

9. A PEG/IL-2 conjugate having the structure:

##STR5##