Compositions relating to a mutant Clostridium difficile toxin and methods thereof

Patent metadata

Field	Value
Publication number	US-RE48862-E
Application number	US-201916387999-A
Country	US
Kind code	E1
Filing date	Apr 18, 2019
Priority date	Apr 22, 2011
Publication date	Dec 28, 2021
Grant date	Dec 28, 2021

Abstract

In one aspect, the invention relates to an immunogenic composition that includes a mutant Clostridium difficile toxin A and/or a mutant Clostridium difficile toxin B. Each mutant toxin includes a glucosyltransferase domain having at least one mutation and a cysteine protease domain having at least one mutation, relative to the corresponding wild-type C. difficile toxin. The mutant toxins may further include at least one amino acid that is chemically crosslinked. In another aspect, the invention relates to antibodies or binding fragments thereof that binds to said immunogenic compositions. In further aspects, the invention relates to isolated nucleotide sequences that encode any of the foregoing, and methods of use of any of the foregoing compositions.

First claim

The invention claimed is:

1. An isolated polypeptide comprising the amino acid sequence set forth in SEQ ID NO: 84, wherein a side chain of a lysine residue of the polypeptide is crosslinked to a betaalanine moiety.

2. The isolated polypeptide according to claim 1 , comprising the amino acid sequence set forth in SEQ ID NO:4, wherein the methionine residue at position 1 is optionally not present.

3. The isolated polypeptide according to claim 1 , further comprising a glycine moiety crosslinked to a side chain of an aspartic acid residue of the polypeptide or to a side chain of a glutamic acid residue of the polypeptide.

4. The isolated polypeptide according to claim 1 , further comprising a crosslink between a second lysine residue of the polypeptide and a side chain of an aspartic acid residue of the polypeptide.

5. The isolated polypeptide according to claim 1 , further comprising a crosslink between a second lysine residue of the polypeptide and a side chain of a glutamic acid residue of the polypeptide.

6. The isolated polypeptide according to claim 1 , wherein the polypeptide has an EC 50 of at least about 100 μg/ml, as measured by an in vitro cytotoxicity assay.

7. The isolated polypeptide according to claim 6 , wherein the polypeptide has an EC 50 of at least about 1000 μg/ml, as measured by an in vitro cytotoxicity assay.

8. An immunogenic composition comprising a first isolated polypeptide having the amino acid sequence set forth in SEQ ID NO: 84 wherein a side chain of a lysine residue of the first polypeptide is crosslinked to a beta-alanine moiety.

9. The immunogenic composition according to claim 8 , further comprising a second isolated polypeptide having the amino acid sequence set forth in SEQ ID NO: 86, wherein a side chain of a lysine residue of the second polypeptide is crosslinked to a beta-alanine moiety.

10. The immunogenic composition according to claim 9 , said first polypeptide comprising the amino acid sequence set forth in SEQ ID NO: 4, wherein the methionine residue at position 1 is optionally not present, and said second polypeptide comprising the amino acid sequence setforth in SEQ ID NO:6.

11. The immunogenic composition according to claim 8 , further comprising a crosslink between a side chain of a second lysine residue of the first polypeptide and a side chain of an aspartic acid residue.

12. The immunogenic composition according to claim 9 , further comprising a crosslink between a side chain of a second lysine residue of the second polypeptide and a side chain of an aspartic acid residue.

13. The immunogenic composition according to claim 8 , further comprising a crosslink between a side chain of a second lysine residue of the first polypeptide and a side chain of a glutamic acid residue.

14. The immunogenic composition according to claim 9 , further comprising a crosslink between a side chain of a second lysine residue of the second polypeptide and a side chain of a glutamic acid residue.

15. The immunogenic composition according to claim 8 , further comprising a crosslink between a side chain of an aspartic acid residue of the first polypeptide and a glycine moiety.

16. The immunogenic composition according to claim 9 , further comprising a crosslink between a side chain of an aspartic acid residue of the second polypeptide and a glycine moiety.

17. The immunogenic composition according to claim 8 , further comprising a crosslink between a side chain of a glutamic acid residue of the first polypeptide and a glycine moiety.

18. The immunogenic composition according to claim 9 , further comprising a crosslink between a side chain of a glutamic acid residue of the second polypeptide and a glycine moiety.

19. The immunogenic composition according to claim 8 , wherein each polypeptide has an EC 50 of at least about 1000 μg/ml, as measured by an in vitro cytotoxicity assay.

20. The immunogenic composition according to claim 8 , further comprising an adjuvant.

Assignees

• Wyeth Llc

Inventors

• Sidhu Maninder K
• Anderson Annaliesa Sybil
• Donald Robert G K
• Jansen Kathrin Ute
• Kalyan Narender K
• Moran Justin Keith
• Ruppen Mark E
• Flint Michael James

Classifications

• A61K39/08 Primary

  Clostridium, e.g. Clostridium tetani · CPC title

• C12N9/1051

  Hexosyltransferases (2.4.1) · CPC title

• C07K2317/92

  Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value · CPC title

• C07K2317/33

  Crossreactivity, e.g. for species or epitope, or lack of said crossreactivity · CPC title

• C07K14/33 Primary

  from Clostridium (G) · CPC title