Biologically active proteins having increased In Vivo and/or In Vitro stability

Abstract

The present invention provides unstructured recombinant polymers (URPs) and proteins containing one or more of the URPs. The present invention also provides microproteins, toxins and other related proteinaceous entities, as well as genetic packages displaying these entities. The present invention also provides recombinant polypeptides including vectors encoding the subject proteinaceous entities, as well as host cells comprising the vectors. The subject compositions have a variety of utilities including a range of pharmaceutical applications.

Description

CROSS-REFERENCE[0001]This application claims the benefit of U.S. Provisional Application No. 60 / 743,410 filed Mar. 6, 2006, which application is incorporated herein by reference. This application is a continuation-in-part application of Ser. Nos. 11 / 528,927 and 11 / 528,950, filed on Sep. 27, 2006, which in turn claim priority to provisional applications Ser. Nos. 60 / 721,270, 60 / 721,188, filed on Sep. 27, 2005 and 60 / 743,622 filed on Mar. 21, 2006, all of which are herein incorporated by reference in their entirety.BACKGROUND OF THE INVENTION[0002]It has been well documented that properties of proteins, in particular plasma clearance and immunogenicity, can be improved by attaching hydrophilic polymers to these proteins (Kochendoerfer, G. (2003) Expert Opin Biol Ther, 3: 1253-61), (Greenwald, R. B., et al. (2003) Adv Drug Deliv Rev, 55: 217-50), (Harris, J. M., et al. (2003) Nat Rev Drug Discov, 2: 214-21). Examples of polymer-modified proteins that have been approved by the FDA for t...

AI Technical Summary

Problems solved by technology

The conjugation step can result in the formation of product mixtures that need to be separated leading to significant product loss.

Such mixtures are difficult to manufacture and characterize and they contain isomers with reduced or no therapeutic activity.

All these methods have significant limitations.

The selective PEGylation of the N-terminus requires careful process control and side reactions are difficult to eliminate.

The introduction of cysteines for PEGylation can interfere with protein production and / or purification.

A further limitation of PEGylation is that PEG is typically manufactured as a mixture of polymers with similar but not uniform length.

The same limitations are inherent in many other chemical polymers.

It has also been demonstrated that oligomeric sequences that are based on such pathogen-derived repetitive sequences can be fused to other proteins resulting in increased serum halflife.

However, these pathogen-derived oligomers have a number of deficiencies.

However, no attempts have been reported to remove T cell epitopes from the sequences contributing to the formation of immune reactions.

Furthermore, the pathogen-derived sequences have not been optimized for pharmacological applications which require sequences with good solubility and a very low affinity for other target proteins.

Images