Compositions relating to a mutant Clostridium difficile toxin and methods thereof

Patent metadata

Field	Value
Publication number	US-RE48863-E
Application number	US-201916388011-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. A composition comprising (a) a first polypeptide, which comprises the amino acid sequence set forth in SEQ ID NO: 4, wherein the methionine residue at position 1 of SEQ ID NO: 4 is not present, wherein a side chain of a lysine residue of the first polypeptide is crosslinked to a beta-alanine moiety, and wherein the first polypeptide further comprises a crosslink between a side chain of an aspartic acid residue of the first polypeptide and a glycine moiety, and a crosslink between a side chain of a glutamic acid residue of the first polypeptide and a glycine moiety; and (b) a second polypeptide, which comprises the amino acid sequence set forth in SEQ ID NO: 6, wherein the methionine residue at position 1 of SEQ ID NO: 6 is not present, wherein a side chain of a lysine residue of the second polypeptide is crosslinked to a beta-alanine moiety, and wherein the second polypeptide further comprises a crosslink between a side chain of an aspartic acid residue of the second polypeptide and a glycine moiety, and a crosslink between a side chain of a glutamic acid residue of the second polypeptide and a glycine moiety.

2. The composition according to claim 1 , wherein the second polypeptide further comprises a dehydroalanine moiety.

3. The composition according to claim 1 , wherein the composition is immunogenic.

4. The composition according to claim 1 , wherein the composition comprises an immunologically effective amount of the first polypeptide and an immunologically effective amount of the second polypeptide.

5. The composition according to claim 1 , wherein the composition is lyophilized.

6. The composition according to claim 1 , wherein the composition further comprises an adjuvant.

7. The composition according to claim 1 , wherein the composition further comprises aluminum hydroxide.

8. The composition according to claim 1 , wherein the composition does not further comprise an adjuvant.

9. The composition according to claim 1 , wherein the composition further comprises trehalose.

10. The composition according to claim 1 , wherein the composition further comprises polysorbate-80.

11. A composition comprising (a) a first polypeptide, which comprises the amino acid sequence set forth in SEQ ID NO: 4, wherein the methionine residue at position 1 of SEQ ID NO: 4 is not present, wherein a side chain of a first lysine residue of the first polypeptide is crosslinked to a beta-alanine moiety, and wherein a second lysine residue of the first polypeptide is crosslinked to an aspartic acid residue or to a glutamic acid residue of the first polypeptide; and (b) a second polypeptide, which comprises the amino acid sequence set forth in SEQ ID NO: 6, wherein the methionine residue at position 1 of SEQ ID NO: 6 is not present, wherein a side chain of a lysine residue of the second polypeptide is crosslinked to a beta-alanine moiety, and wherein a second lysine residue of the second polypeptide is crosslinked to an aspartic acid residue or to a glutamic acid residue of the second polypeptide.

12. The composition according to claim 11 , wherein the second polypeptide further comprises a dehydroalanine moiety.

13. The composition according to claim 11 , wherein the composition is immunogenic.

14. The composition according to claim 11 , wherein the composition comprises an immunologically effective amount of the first polypeptide and an immunologically effective amount of the second polypeptide.

15. The composition according to claim 11 , wherein the composition is lyophilized.

16. The composition according to claim 11 , wherein the composition further comprises an adjuvant.

17. The composition according to claim 11 , wherein the composition further comprises aluminum hydroxide.

18. The composition according to claim 11 , wherein the composition does not further comprise an adjuvant.

19. The composition according to claim 11 , wherein the composition further comprises trehalose.

20. The composition according to claim 11 , wherein the composition further comprises polysorbate-80.

Assignees

• Wyeth Llc

Inventors

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

Classifications

• C12N9/1051

  Hexosyltransferases (2.4.1) · CPC title

• A61K39/08 Primary

  Clostridium, e.g. Clostridium tetani · CPC title

• A61K39/085

  Staphylococcus · CPC title

• C07K16/1282

  Clostridium (G) · CPC title

• C12N9/99

  Enzyme inactivation by chemical treatment · CPC title