SYNTHESIS OF LIPOPOLYSACCHARIDE-PROTEIN CONJUGATE VACCINES VIA THE LIPID A REGION FOLLOWING REMOVAL OF THE GLYCOSIDIC PHOSPHATE RESIDUE
(19)AUSTRALIAN PATENT OFFICE (54) Title SYNTHESIS OF LIPOPOLYSACCHARIDE-PROTEIN CONJUGAI b VACCINES VIA THE LIPID A REGION FOLLOWING REMOVAL OF THE GLYCOSIDIC PHOSPHATE RESIDUE (51)G International Patent Classification(s) AG IK 047/48 A61K 039/02 (21) Application No: 2003206532 (22) Application Date: 2003.02.24 (87) WIPONo: WO03/070282 (30) Priority Data (31) Number (32) Date 60/358,384 2002.02.22 (33) Country US 7 (43) Publication Date : 2003.09.09 (43) Publication Journal Date : 2003.1 0.09 (71) Applicant(s) NATIONAL RESEARCH COUNCIL OF CANADA (72) Inventor(s) RICHARDS, James, C; COX, Andrew, MOXON, Richard; JENNINGS, Harold; MIESZALA Malgorzata; KOGAN, Grigorii; ZOU, Wei fM) Application NoAU2003206532 A1(19)AUSTRALIAN PATENT OFFICE (54) Title SYNTHESIS OF LIPOPOLYSACCHARIDE-PROTEIN CONJUGAI b VACCINES VIA THE LIPID A REGION FOLLOWING REMOVAL OF THE GLYCOSIDIC PHOSPHATE RESIDUE (51)G International Patent Classification(s) AG IK 047/48 A61K 039/02 (21) Application No: 2003206532 (22) Application Date: 2003.02.24 (87) WIPONo: WO03/070282 (30) Priority Data (31) Number (32) Date 60/358,384 2002.02.22 (33) Country US 7 (43) Publication Date : 2003.09.09 (43) Publication Journal Date : 2003.1 0.09 (71) Applicant(s) NATIONAL RESEARCH COUNCIL OF CANADA (72) Inventor(s) RICHARDS, James, C; COX, Andrew, MOXON, Richard; JENNINGS, Harold; MIESZALA Malgorzata; KOGAN, Grigorii; ZOU, Wei This invention relates to lipopolysaccharide-protein conjugate vaccines with appropriate presentation of conserved inner core oligosaccharide epitopes having improved immunogenic properties. These are based upon antigenic, detoxified bacterial lipopolysaccharides which optimally present an inner core oligosaccharide epitope following removal of at least a glycosidic phosphate of the lipid A region. These partially or completely dephosphorylated antigenic, detoxified bacterial lipopolysaccharides are linkable to an immunologically acceptable carrier and can be used in polyvalent or multivalent vaccines. CLAIMS : 1. A method of linking an antigenic, detoxified bacterial lipopolysaccharide having a lipid A region with a terminal glucosamine glycosidic phosphate group to an immunologically acceptable carrier through said lipid A region, which method comprises removing said terminal glycosidic phosphate group to yield a partially or completely dephosphorylated detoxified bacterial lipopolysaccharide and then conjugating said partially or completely dephosphorylated detoxified bacterial lipopolysaccharide to said immunologically acceptable carrier.
2. An antigenic, partially or completely dephosphorylated detoxified bacterial lipopolysaccharide linkable to an immunologically acceptable carrier through complete dephosphorylation of glycosidically-linked phosphate or phosphate substituents of glycose at the reducing terminus in the lipid A region.
3. The antigenic, partially or completely dephosphorylated detoxified bacterial lipopolysaccharide of claim 2 comprising an oligosaccharide epitope.
4. The antigenic, partially or completely dephosphorylated detoxified bacterial lipopolysaccharide of claim 3 wherein said oligosaccharide epitope is conserved.
5. The antigenic, partially or completely dephosphorylated detoxified bacterial lipopolysaccharide of claim 3 wherein said oligosaccharide epitope is a conserved inner core oligosaccharide epitope.
6. A conjugate vaccine for combating a Gram-negative or other bacterium comprising an antigenic, partially or completely dephosphorylated detoxified bacterial lipopolysaccharide according to any one of claims 2 to 5 linked to an immunologically acceptable carrier through complete dephosphorylation of glycosidically-linked phosphate or phosphate substituents of glycose at the reducing terminus in the lipid A region. <Desc/Clms Page number 43> 7. The conjugate vaccine of claim 6, wherein said immunogenic carrier is a protein.
8. The vaccine of claim 7, wherein said immunogenic carrier protein is selected from the group consisting of tetanus toxin/toxoid, cross-reacting material (CRM), NTHi high molecular weight protein, diphtheria toxin/toxoid, detoxified P. aeruginosa toxin A, cholera toxin/toxoid, pertussis toxin/toxoid, Clostridium perfringens exotoxins/toxoid, hepatitis B surface antigen, hepatitis B core antigen, rotavirus VP 7 protein, respiratory syncytial virus F and G proteins.
9. The vaccine of claim 8, wherein said immunogenic carrier protein is tetanus toxoid.
10. The vaccine of claim 8, wherein said immunogenic carrier protein is CRM197 11. A conjugate vaccine for combating a Gram-negative bacterium comprising an antigenic, partially or completely dephosphorylated detoxified bacterial lipopolysaccharide according to any one of claims 2 to 5 linked via a linker to an immunologically acceptable carrier through complete dephosphorylation of glycosidically-linked phosphate or phosphate substituents of glycose at the reducing terminus in the lipid A region.
12. The vaccine of claim 11, wherein said linker is selected from the group consisting of M2C2H., cystamine, adipic acid di-hydrazide, s-aminohexanoic acid, chlorohexanol dimethyl acetal, D-glucuronolactone and p- nitrophenylethyl amine.
13. The vaccine of claim 12, wherein said linker is M2C2H.
14. The vaccine of claim 12, wherein said linker is cystamine. <Desc/Clms Page number 44> 15. A pharmaceutical composition comprising the conjugate vaccine of claims 6 or 11 in association with an adjuvant.
16. The pharmaceutical composition of claim 15, wherein said adjuvant is selected from the group consisting of Freund's adjuvant, alum and Ribi.
17. The pharmaceutical composition of claim 15 or 16 further comprising a pharmaceutical acceptable diluent.
18. A polyvalent conjugate vaccine comprising a plurality of different antigenic, detoxified bacterial lipopolysaccharides according to claim 2 covalently linked to an immunogenic carrier.
19. A multivalent conjugate vaccine comprising a plurality of different conjugate vaccines according to claims 6 or 11.