Modified polypeptides

Abstract

The present invention relates to polypeptides with reduced immune response including reduced allergenicity having one or more amino acid residues being substituted with other amino acid residues and / or having coupled one or more polymeric molecules in the vicinity of the polypeptides metal binding site, a method for preparing modified polypeptides of the invention, the use of the polypeptide for reducing the immunogenicity and allergenicity and compositions comprising the polypeptide.

Description

CROSS-REFERENCES TO RELATED APPLICATIONS[0001] This application is a continuation of application Ser. No. 09 / 417,359 filed Oct. 13, 1999 and claims priority under 35 U.S.C. 119 of U.S. provisional application Nos. 60 / 105,624 and 60 / 157,426 filed on Oct. 26, 1998 and Oct. 4, 1999, respectively, and of Danish application nos. PA 1998 01301 and PA 1999 01418 filed on Oct. 13, 1998 and Oct. 4, 1999, respectively, the contents of which are fully incorporated herein by reference.[0002] The present invention relates to polypeptides having substituted one or more amino acid residues to the polypeptide and / or having coupled polymeric molecules on the surface of the 3-dimensional structure of the polypeptide, a method for preparing modified polypeptides of the invention, the use of the modified polypeptides for reducing immunogenicity and allergenicity, and compositions comprising the polypeptide.DESCRIPTION OF THE RELATED ART[0003] The use of polypeptides, including enzymes, in the circulato...

AI Technical Summary

Benefits of technology

[0017] The present inventors have found that polypeptides, such as enzymes, may be made less immunogenic and / or allergenic by substituting one or more amino acid residues on the surface of the polypeptide with other amino acid residues and / or by coupling polymeric molecules on the surface of the enzyme in the vicinity of a bound ligand of the enzyme e.g. a metal ion substantially without affecting the enzymatic activity.

[0022] According to the present invention it is possible to provide polypeptides with a reduced immune response, which has a substantially retained residual activity.

[0040] The invention also relates to the use of a modified polypeptide of the invention and the method of the invention for reducing the immunogenicity of pharmaceuticals and reducing the allergenicity of industrial products.

Problems solved by technology

Certain polypeptides and enzymes have an unsatisfactory stability and may under certain circumstances--dependent on the way of challenge--cause an immune response, typically an IgG and / or IgE response.

It has been found that the attachment of polymeric molecules to a polypeptide often has the effect of reducing the activity of the polypeptide by interfering with the interaction between the polypeptide and its substrate.

The potential risk in connection with industrial polypeptides is inhalation causing an allergenic response in the form of mainly IgE antibody formation.

Therefore, in connection with industrial polypeptides the potential risk is respiratory allergenicity caused by inhalation, intratracheal and intranasal presentation of polypeptides.

The main potential risk of pharmaceutical polypeptides is immunogenicity caused by intradermal, intravenous or subcutaneous presentation of the polypeptide.

For enzymes, there is a conflict between reducing the immune response and maintaining a substantial residual enzymatic activity.

Without being limited to any theory it is believed that the loss of enzymatic activity of enzyme-polymer conjugates might be a consequence of impeded access of the substrate to the active site in the form of spatial hindrance of the substrate by especially bulky and / or heavy polymeric molecules to the catalytic cleft.

It might also, at least partly, be caused by disadvantageous minor structural changes of the 3-dimensional structure of the enzyme due to the stress made by the coupling of the polymeric molecules.

Therefore, in the case of the industrial polypeptide the potential risk is respiratory allergy (i.e. IgE response) as a consequence of inhalation of polypeptides through the respiratory passage.

Coupling polymeric molecules to hydroxy groups are generally very difficult as it must be performed in water.

The reactivity of these compounds are very high but may make the hydrolysis too fast.

Regions containing large inserts (.gtoreq.3 residues) relative to the known 3-dimensional structures are known to be quite difficult to model, and structural predictions must be considered with care.

In that case, it may be difficult to assess a priori how well the functionality of the protein is maintained while antigenicity, immunogenicity and / or allergenicity is reduced.

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