Combined enteropathogen recombinant construct

Patent metadata

Field	Value
Publication number	US-10105448-B2
Application number	US-201514721656-A
Country	US
Kind code	B2
Filing date	May 26, 2015
Priority date	Jan 11, 2005
Publication date	Oct 23, 2018
Grant date	Oct 23, 2018

Abstract

The inventive subject matter relates to a construct comprising antigens derived from multiple enterobacteria including Campylobacter jejuni capsule polysaccharide polymer, enterotoxigenic Escherichia coli recombinant polypeptide construct and lipopolysaccharide from Shigella spp. The subject invention also relates to a method of inducing an immune response utilizing the inventive composition.

First claim

What is claimed is:

1. A multi-agent immunogenic construct, comprising a Campylobacter jejuni capsule polysaccharide conjugated to a protein carrier, wherein said protein carrier comprises an Escherichia coli enterotoxigenic recombinant polypeptide construct, wherein said Escherichia coli recombinant polypeptide construct comprises a minor or major subunit connected to one or more major fimbrial subunits or immunogenic fragments, thereof, of the same fimbrial type, via a polypeptide linker, are also connected to each other via a polypeptide linker wherein the C-terminal Escherichia coli fimbrial major subunit is connected, via a linker, to a C-terminal donor β strand derived from a major Escherichia coli fimbrial subunit that is homologous or heterologous to the immediately N-terminal major subunit, wherein said donor β strand contains 12 to 16 amino acids from the N-terminal region of said one or more major fimbrial subunits, and wherein said recombinant polypeptide construct can be connected to one or more additional recombinant polypeptide constructs, wherein each of the constructs contain fimibrial subunits derived from a different fimbrial type than any of the other constructs and wherein the recombinant polypeptide construct can contain a C-terminal histidine tag at the C-terminus, wherein said minor or major Escherichia coli fimbrial subunits are derived from ETEC strains selected from the group consisting of Class 5, CS3 and CS6.

2. The multi-agent immunogenic construct of claim 1 , wherein the multi-agent immunogenic construct comprises Shigella lipopolysaccharide in place of or in addition to the Campylobacter jejuni capsule polysaccharide.

3. The multi-agent immunogenic construct of claim 1 , wherein said molar ratio of Campylobacter jejuni or Shigella spp. lipopolysaccharide to Escherichia coli recombinant protein carrier is 1:1 to 5:1.

4. The multi-agent immunogenic construct of claim 1 , wherein the Escherichia coli recombinant polypeptide construct comprises the amino acid sequence selected from the group consisting of SEQ ID Nos. 108 and 110.

5. The multi-agent immunogenic construct of claim 1 , wherein the Escherichia coli recombinant polypeptide construct is encoded by the nucleotide sequence of SEQ ID Nos. 105 and 109.

6. The multi-agent immunogenic construct of claim 1 , wherein said Campylobacter jejuni polysaccharide is a repeating trisaccharide structure having the formula selected from the group consisting of:
[→3)-α-D-Gal-(1→2)-6d-α-D-altro-Me-Hep-(1→3)-β-D-GlcNAc-(1→] n ;
[→3)-β-6-deoxy-D-ido-Heptose(1→4)-β-D-GlcNAc(1→] n ;
selected from the group consisting of:

[→3)-L-beta-ido-Hep-(1→4)-beta-D-Glc-(1→] n , with non-stoichiometric substitution of O-methyl-phosphoramidate at position 2 of L-glycero-beta-D-ido-heptose;
[→3)-6d-beta-ido-Hep-(1→4)-beta-D-Glc-(1→] n , derived from HS13, with non-stoichiometric substitution of O-methyl-phosphoramidate at position 2 or/and 7 of 6-deoxy-beta-D-ido-heptose;
[→3)-L-beta-D-ido-Hep-(1→4)-beta-D-Glc-(1→] n ;
[→3)-L-alpha-D-ido-Hep-(1→4)-alpha-Gal-(1→] n , with non-stoichiometric substitution of O-methyl-phosphoramidate at position 2 of 6-deoxy-alpha-D-ido-heptose;

[→3)-6d-alpha-D-ido-Hep-(1→4)-alpha-Gal-(1→] n , derived from HS3, HS13 and HS50 with non-stoichiometric substitution of O-methyl-phosphoramidate at position 2 of L-glycero-alpha-D-ido-heptose, wherein “n” is 1 to 100.

7. The multi-agent immunogenic construct of claim 2 , wherein said Shigella lipopolysaccharide has the structure:

8. The multi-agent immunogenic construct of claim 1 , wherein said N-terminus of said minor or major subunit contains an 18-22 amino acid signal peptide.

9. The multi-agent immunogenic construct of claim 1 , wherein the amino acid sequence of said polypeptide linker is the amino acid sequence of SEQ ID No. 5 or a tri-glycine.

10. The multi-agent immunogenic construct of claim 1 , wherein one or more major subunits contain a deletion of the 14 to 18 N-terminal amino acids.

11. The multi-agent Immunogenic construct of claim 1 , wherein said amino acid sequence of said Escherichia coli fimbrial minor subunit is selected from the group consisting of SEQ ID Nos. 46, 52 and 58, or derivatives thereof, and wherein said amino acid sequence of said Escherichia coli fimbrial major subunit is selected from the group consisting of SEQ ID Nos. 55 and 61, or derivatives thereof.

12. A method of inducing an immune response against C. jejuni strains, wherein said method induces an immune response against one or more enterobacteria selected from the group consisting of C. jejuni strains, Escherichia coli , and Shigella , comprising the steps: a. administering the multi-agent immunogenic composition of claim 1 at a dose range of 0.1 μg to 10 mg per dose; b. administering a boosting dose of said capsule polysaccharide composition as in section a) at a dose range of 0.1 μg to 10 mg per dose.

13. The method of claim 12 , wherein said Campylobacter jejuni capsule polysaccharide comprises the polysaccharide structures of claim 6 .

14. The method of claim 12 , wherein said multi-agent immunogenic composition comprises the construct of claim 2 .

15. The method of claim 12 , wherein said Escherichia coli enterotoxigenic recombinant polypeptide construct wherein said Escherichia coli fimbrial minor subunit is selected from the group consisting of SEQ ID Nos. 46 and 58 or derivatives thereof, and wherein said amino acid sequence of said Escherichia coli fimbrial major subunit is selected from the group consisting of SEQ ID Nos. 49, 55 and 61, or derivatives thereof.

16. The method of claim 12 , wherein the Escherichia coli recombinant polypeptide construct comprises the amino acid sequence selected from the group consisting of SEQ ID Nos. 108 and 110.

17. The method of claim 12 , wherein the Escherichia coli recombinant polypeptide construct is encoded by the nucleotide sequence of SEQ ID Nos. 105 and 109.

18. The method of claim 12 , wherein said N-terminus of said minor or major subunit contains an 18-22 amino acid signal peptide.

19. The method of claim 12 , wherein the amino acid sequence of said polypeptide linker is the amino acid sequence of SEQ ID No. 5 or a tri-glyicine.

20. The method of claim 12 , wherein one or more major subunits contain a deletion of the 14 to 18 N-terminal amino acids.

Assignees

• Guerry Patricia
• Monteiro Mario Artur
• Savarino Stephen
• Us Navy

Inventors

• Guerry Patricia
• Monteiro Mario Artur
• Savarino Stephen

Classifications

• Y02A50/47

  Cross-Sectional Technologies · mapped topic

• A61K39/0283

  Shigella · CPC title

• A61K2039/55505

  Inorganic adjuvants · CPC title

• C07K16/1232

  Escherichia (G) · CPC title

• C07K2319/35

  containing a fusion for enhanced stability/folding during expression, e.g. fusions with chaperones or thioredoxin · CPC title