Derivatives of Growth Hormone and Related Proteins, and Methods of Use Thereof

Abstract

The growth hormone supergene family comprises greater than 20 structurally related cytokines and growth factors. A general method is provided for creating site-specific, biologically active conjugates of these proteins. The method involves adding cysteine residues to non-essential regions of the proteins or substituting cysteine residues for non-essential amino acids in the proteins using site-directed mutagenesis and then covalently coupling a cysteine-reactive polymer or other type of cysteine-reactive moiety to the proteins via the added cysteine residue. Disclosed herein are preferred sites for adding cysteine residues or introducing cysteine substitutions into the proteins, and the proteins and protein derivatives produced thereby. Also disclosed are therapeutic methods for using the cysteine variants of the invention.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS [0001] This application is a continuation under 35 U.S.C. § 120 of U.S. application Ser. No. 10 / 685,288, filed Oct. 13, 2003, which claims the benefit of priority under 35 U.S.C. 119(e) from U.S. Provisional Application Ser. No. 60 / 418,106, filed Oct. 11, 2002, entitled “Methods of Use for Granulocyte Colony Stimulating Factor Cysteine Muteins”, and from U.S. Provisional Application Ser. No. 60 / 418,105, filed Oct. 11, 2002, entitled “Methods of Use for Erythropoietin Cysteine Muteins”. [0002] U.S. application Ser. No. 10 / 685,288, supra, is also a continuation-in-part of U.S. application Ser. No. 10 / 400,377, filed Mar. 26, 2003, which is a divisional of U.S. application Ser. No. 09 / 462,941, filed Jan. 14, 2000, now U.S. Pat. No. 6,608,183, which is a national stage filing under 35 U.S.C. 371 of PCT Application Serial No. PCT / US98 / 14497, filed Jul. 13, 1998, which designates the United States, was published in English, and claims the benefit of ...

AI Technical Summary

Benefits of technology

[0036] Also described herein are therapeutic methods for inhibiting growt

Problems solved by technology

Proteins are expensive to manufacture and, unlike conventional small molecule drugs, are not readily absorbed by the body.

Moreover, they are digested if taken orally.

Patients dislike injections, which leads to reduced compliance and reduced drug efficacy.

However, glycosylated proteins are produced using expensive mammalian cell expression systems.

Similar problems plague protein antagonists.

These fluctuations can lead to decreased efficacy and increased frequency of adverse side effects for protein therapeutics.

The rapid clearance of recombinant proteins from the body significantly increases the amount of protein required per patient and dramatically increases the cost of treatment.

The cost of human protein pharmaceuticals is expected to increase dramatically in the years ahead as new and existing drugs are approved for more disease indications.