Beta-lactamases with improved properties for therapy
This invention relates to, in part, compositions of beta-lactamases and methods of using these enzymes in, for example, gastrointestinal tract (GI tract) disorders such as C. difficile infection (CDI).
1. A beta-lactamase comprising an amino acid sequence having at least 90% sequence identity with SEQ ID NO: 1 and having a D276N and one or more of F33Y, S240P, T243I, and S266N mutations according to Ambler classification.
2. The beta-lactamase of claim 1 , comprising a D276N mutation and a F33Y mutation according to Ambler classification.
3. The beta-lactamase of claim 2 , further comprising a S240P mutation according to Ambler classification.
4. The beta-lactamase of claim 1 , comprising a D276N mutation and a T243I mutation according to Ambler classification.
5. The beta-lactamase of claim 4 , further comprising a S266N mutation according to Ambler classification.
6. The beta-lactamase of claim 1 , wherein the beta-lactamase hydrolyzes one or more of penicillins and cephalosporins.
7. The beta-lactamase of claim 6 , wherein the penicillin is ampicillin.
8. The beta-lactamase of claim 6 , wherein the cephalosporin is selected from ceftriaxone, cefotaxime, cefozolin, cefoperazone, cefepime, cefuroxime, and ceftazidime.
9. The beta-lactamase of claim 1 , wherein the beta-lactamase has improved enzymatic activity against a cephalosporin as compared to SEQ ID NO: 1.
10. The beta-lactamase of claim 9 , wherein the cephalosporin is ceftriaxone.
11. A polynucleotide comprising a polynucleotide sequence encoding the beta-lactamase of claim 1 .
12. A host cell comprising the polynucleotide of claim 11 .
13. A pharmaceutical composition, comprising the beta-lactamase of claim 1 and a pharmaceutically acceptable carrier or excipient.
14. The pharmaceutical composition of claim 13 , wherein the composition is formulated for oral administration, optionally selected from a tablet, a multi-particulate sprinkle, and a multi-particulate capsule.
15. A method for preventing an antibiotic-induced adverse effect in the GI tract, comprising administering an effective amount of a beta-lactamase to a patient in need thereof, wherein the beta-lactamase comprises an amino acid sequence having at least 90% sequence identity with SEQ ID NO: 1 and having a D276N and one or more of F33Y, S240P, T243I, and S266N mutations according to Ambler classification.
16. The method of claim 15 , wherein the beta-lactamase comprises a D276N mutation and a F33Y mutation according to Ambler classification.
17. The method of claim 16 , wherein the beta-lactamase further comprises a S240P mutation according to Ambler classification.
18. The method of claim 15 , wherein the beta-lactamase comprises a D276N mutation and a T243I mutation according to Ambler classification.
19. The method of claim 18 , wherein the beta-lactamase further comprises a S266N mutation according to Ambler classification.
20. A method for treating or preventing an antibiotic induced C. difficile infection (CDI) and/or a C. difficile -associated disease, comprising administering an effective amount of a beta-lactamase to a patient in need thereof, wherein the beta-lactamase comprises an amino acid sequence having at least 90% sequence identity with SEQ ID NO: 1 and having a D276N and one or more of F33Y, S240P, T243I, and S266N mutations according to Ambler classification.
21. The method of claim 20 , wherein the beta-lactamase comprises a D276N mutation and a F33Y mutation according to Ambler classification.
22. The method of claim 21 , wherein the beta-lactamase further comprises a S240P mutation according to Ambler classification.
23. The method of claim 20 , wherein the beta-lactamase comprises a D276N mutation and a T243I mutation according to Ambler classification.
24. The method of claim 23 , wherein the beta-lactamase further comprises a S266N mutation according to Ambler classification.
25. The method of claim 20 , wherein the C. difficile -associated disease is antibiotic-associated diarrhea.