E. coli FimH mutants and uses thereof

Patent metadata

Field	Value
Publication number	US-12357681-B2
Application number	US-202117554424-A
Country	US
Kind code	B2
Filing date	Dec 17, 2021
Priority date	Dec 23, 2020
Publication date	Jul 15, 2025
Grant date	Jul 15, 2025

Abstract

This disclosure relates to the design of E. coli mutated FimH polypeptides that result in improved biochemical properties and immunogenicity, compositions comprising such polypeptides, and uses thereof.

First claim

The invention claimed is:

1. A mutated FimH polypeptide, which comprises: at least one amino acid substitution relative to the amino acid sequence of the wild-type FimH polypeptide, wherein the substitution is selected from the group consisting of: F1I; F1V; F1M; F1Y; F1W; P12C; G14C; G15A; GI5P; G16A; G16P; A18C; P26C; V28C; Q32C; N33C; L34N; L34S; L34T; L34D; L34E; L34K; L34R; V35C; S62C; Y64C; G65A; L68C; F71C; T86C; L107C; Y108C; L109C; V112C; S113C; A115V; G116C; V118C; A119C; A119N; A119S; A119D; A119E; A119K; A119R; A127C; L129C; F144C; V154C; V155C; V156C; T158C; and V185I, and wherein the amino acid positions are numbered according to SEQ ID NO: 59.

2. A mutated FimH polypeptide, which comprises amino acid substitutions relative to the amino acid sequence of the wild-type FimH polypeptide, wherein the amino acid substitutions selected from the group consisting of:
a) G15A and G16A;
b) P12C and A18C;
c) G14C and F144C;
d) P26C and V35C;
e) P26C and V154C;
f) P26C and V156C;
g) V27A and Q133K;
h) V28C and N33C;
i) V28C and P157C;
j) Q32C and Y108C;
k) N33C and L109C;
I) N33C and P157C;
m) V35C and L107C;
n) V35C and L109C;
o) S62C and T86C;
p) S62C and L129C;
q) Y64C and L68C;
r) Y64C and A127C;
s) L68C and F71C;
t) V112C and T158C;
u) S113C and G116C;
v) S113C and T158C;
w) V118C and V156C;
x) A119C and V155C;
y) L34N and V27A;
z) L34S and V27A;
aa) L34T and V27A;
ab) L34D and V27A;
ac) L34E and V27A;
ad) L34K and V27A;
ae) L34R and V27A;
af) A119N and V27A;
ag) A119S and V27A;
ah) A119T and V27A;
ai) A119D and V27A;
aj) A119E and V27A;
ak) A119K and V27A;
al) AI119R and V27A;
am) G15A and V27A;
an) G16A and V27A;
ao) G15P and V27A;
ap) GI6P and V27A;
aq) G15A, G16A, and V27A;
ar) G65A and V27A; and
as) G15A, G16A, V27A, and Q133K, and wherein the amino acid positions are numbered according to SEQ ID NO: 59.

3. A mutated FimH polypeptide comprising a sequence selected from the group consisting of SEQ ID NOs: 2, 3, 5-8, 10-14, 16-21, 23-46, 48-58, and 61-64.

4. The mutated FimH polypeptide according to claim 1 , wherein the polypeptide is isolated.

5. A pharmaceutical composition comprising (i) the mutated FimH polypeptide according to claim 1 and (ii) a pharmaceutically acceptable carrier.

6. An immunogenic composition comprising the mutated FimH polypeptide according to claim 1 .

7. The immunogenic composition according to claim 6 , further comprising an E. coil saccharide antigen, wherein the saccharide antigen comprises a structure selected from the group consisting of formula O25b, Formula O1A, Formula O2, and formula O6.

8. The immunogenic composition according to claim 6 , further comprising at least one adjuvant.

9. The polypeptide of claim 1 , further comprising the amino acid substitution V27A.

10. The polypeptide of claim 2 , comprising the amino acid substitutions G15A, G16A, and V27A.

11. The polypeptide of claim 1 , wherein the polypeptide is stabilized by a donor-strand peptide of FimG (FimH-DsG).

12. The mutated polypeptide of claim 3 , comprising the sequence of SEQ ID NO: 62.

13. A pharmaceutical composition comprising (i) the mutated FimH polypeptide according to claim 2 and (ii) a pharmaceutically acceptable carrier.

14. An immunogenic composition comprising the mutated FimH polypeptide according to claim 2 .

15. The immunogenic composition according to claim 14 , further comprising an E. coli saccharide antigen, wherein the saccharide antigen comprises a structure selected from the group consisting of Formula O25b, Formula O1A, Formula O2, and Formula O6.

16. The immunogenic composition according to claim 14 , further comprising at least one adjuvant.

17. A pharmaceutical composition comprising (i) the mutated FimH polypeptide according to claim 3 and (ii) a pharmaceutically acceptable carrier.

18. An immunogenic composition comprising the mutated FimH polypeptide according to claim 3 .

19. The immunogenic composition according to claim 18 , further comprising an E. coli saccharide antigen, wherein the saccharide antigen comprises a structure selected from the group consisting of Formula O25b, Formula O1A, Formula O2, and Formula O6.

20. The immunogenic composition according to claim 18 , further comprising at least one adjuvant.

Assignees

• Pfizer

Inventors

• Che Ye
• Chorro Laurent Oliver
• Donald Robert George Konrad
• Griffor Matthew Curtis
• Silmon De Monerri Natalie Clare

Classifications

• C07K16/1232

  Escherichia (G) · CPC title

• C07K14/245 Primary

  Escherichia (G) · CPC title

• A61K2039/55566

  Emulsions, e.g. Freund's adjuvant, MF59 · CPC title

• A61K39/0266

  Klebsiella · CPC title

• A61P31/04

  Antibacterial agents · CPC title