Conjugation of Staphylococcus aureus type 5 and type 8 capsular polysaccharides

Patent metadata

Field	Value
Publication number	US-8974799-B2
Application number	US-201013499238-A
Country	US
Kind code	B2
Filing date	Sep 30, 2010
Priority date	Sep 30, 2009
Publication date	Mar 10, 2015
Grant date	Mar 10, 2015

Abstract

The invention provides a process for preparing a conjugate of a S. aureus type 5 or type 8 capsular polysaccharide and a carrier molecule, comprising the steps of: (a) depolymerizing the capsular polysaccharide, to give a polysaccharide fragment; (b) oxidizing the fragment in order to introduce an aldehyde group into at least one saccharide residue in the fragment, to give an oxidized saccharide residue; and (c) coupling the oxidized saccharide residue to a carrier molecule via the aldehyde group, thereby giving the conjugate. The coupling in step (c) may be direct, or may be via a linker molecule. The invention also provides a conjugate obtained or obtainable by this process.

First claim

The invention claimed is:

1. A process of preparing a conjugate of an isolated fragment of the type 5 capsular polysaccharide of Staphylococcus aureus and a carrier molecule comprising the steps of (a) depolymerising an isolated full-length type 5 capsular polysaccharide of S. aureus to provide an isolated fragment of the type 5 capsular polysaccharide having a molecular mass of between 5 kDa and 100 kDa; (b) oxidising the fragment of the type 5 capsular polysaccharide in order to introduce an aldehyde group into at least one saccharide residue in the fragment to have at least one oxidised saccharide residue in the fragment; and (c) coupling the fragment having the at least one oxidised saccharide residue to the carrier molecule via the aldehyde group, thereby preparing the conjugate.

2. A process of producing a purified fragment of a type 5 capsular polysaccharide of Staphylococcus aureus comprising the step of depolymerizing a purified full-length type 5 S. aureus capsular polysaccharide by acid hydrolysis under conditions suitable for the depolymerization, thereby producing the fragment of the purified type 5 capsular polysaccharide of S. aureus , wherein the fragment has a □-D-FucNAc-(1→moiety at its non-reducing terminus.

3. The process of claim 2 , wherein the acid hydrolysis comprises treating the full-length purified type 5 S. aureus capsular polysaccharide with 2% (v/v) acetic acid at 90° C. for 3 hours or overnight.

4. The process of claim 1 , wherein the depolymerising is carried out by acid hydrolysis using acetic acid.

5. The process of claim 1 , wherein the fragment has a degree of 0-acetylation of 10% to 90%.

6. The process of claim 1 , wherein the step (a) comprises depolymerising the full-length type 5 S. aureus capsular polysaccharide to provide the fragment of the type 5 S. aureus capsular polysaccharide having a □-D-FucNAc-(1→moiety at its non-reducing terminus.

7. The process of claim 6 , wherein the step (b) comprises oxidising the fragment of the type 5 S. aureus capsular polysaccharide having the □-D-FucNAc-(1→moiety at its non-reducing terminus to convert the two vicinal hydroxyl groups in the □-D-FucNAc-(1→moiety into two aldehyde groups.

8. A method of providing an oxidised Staphylococcus aureus type 5 capsular polysaccharide comprising the step of oxidising an isolated S. aureus type 5 capsular polysaccharide having a □-D-FucNAc-(1→moiety at its non-reducing terminus to convert the two vicinal hydroxyl groups in the □-D-FucNAc-(1→moiety into two aldehyde groups.

9. A process of providing a coupled Staphylococcus aureus type 5 capsular polysaccharide comprising the step of coupling to a carrier molecule an isolated S. aureus type 5 capsular polysaccharide having a □-D-FucNAc-(1→moiety at its non-reducing terminus that has been oxidised to convert its two vicinal hydroxyl groups into two aldehyde groups, wherein the coupling is via one of the aldehyde groups.

10. The process of claim 1 , wherein the coupling is direct coupling by reacting the aldehyde group with an amine group in the carrier molecule by reductive amination.

11. The process of claim 1 , wherein the coupling is via a linker by reacting the aldehyde group with an amine group in the linker by reductive amination.

12. The process of claim 11 , wherein the linker is already attached to the carrier molecule.

13. The process of claim 1 , wherein the carrier molecule is a carrier protein and wherein the coupling results in a ratio of the fragment of the type 5 S. aureus capsular polysaccharide to the carrier protein (w/w) of between 1:5 and 1:2.

14. The process of claim 11 , wherein the linker attached to the fragment of the capsular polysaccharide is then attached to the carrier molecule.

Assignees

• Berti Francesco
• Costantino Paolo
• Romano Maria Rosaria
• Novartis Ag

Inventors

• Berti Francesco
• Costantino Paolo
• Romano Maria Rosaria

Classifications

• A61P37/04

  Immunostimulants · CPC title

• A61P43/00

  Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00 · CPC title

• A61P31/00

  Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics · CPC title

• A61P31/04

  Antibacterial agents · CPC title

• A61K39/085

  Staphylococcus · CPC title