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 A232G, A237S, A238G, S240D, and D276N mutations according to Ambler classification.
2. The beta-lactamase of claim 1 , wherein the beta-lactamase hydrolyzes one or more of penicillins and cephalosporins.
3. The beta-lactamase of claim 2 , wherein the penicillin is ampicillin.
4. The beta-lactamase of claim 2 , wherein the cephalosporin is selected from ceftriaxone, cefotaxime, cefozolin, cefoperazone, cefepime, cefuroxime, and ceftazidime.
5. The beta-lactamase of claim 1 , wherein the beta-lactamase has improved catalytic efficiency against cephalosporins as compared to SEQ ID NO:1.
6. The beta-lactamase of claim 5 , wherein the beta-lactamase has about a 10 fold to about a 1000 fold improved catalytic efficiency against a cephalosporin as compared to SEQ ID NO:1.
7. The beta-lactamase of claim 6 , wherein the cephalosporin is selected from ceftriaxone, cefotaxime, cefozolin, cefoperazone, cefepime, cefuroxime, and ceftazidime.
8. The beta-lactamase of claim 1 , wherein the beta-lactamase has improved catalytic efficiency against a cephalosporin as compared to P3A.
9. The beta-lactamase of claim 8 , wherein the cephalosporin is selected from cefotaxime, cefepime, and ceftriaxone.
10. The beta-lactamase of claim 1 , wherein the beta-lactamase is active in the GI tract.
11. The beta-lactamase of claim 10 , wherein the beta-lactamase is stable in the small intestine, optionally selected from one or more of the duodenum, jejunum, and ileum.
12. A polynucleotide comprising a polynucleotide sequence encoding the beta-lactamase of claim 1 .
13. A host cell comprising the polynucleotide of claim 12 .
14. A pharmaceutical composition, comprising the beta-lactamase of claim 1 and a pharmaceutically acceptable carrier or excipient.
15. The pharmaceutical composition of claim 14 , wherein the composition is formulated for oral administration, optionally selected from a tablet, multi-particulate sprinkle, and a multi-particulate capsule.
16. 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 A232G, A237S, A238G, S240D, and D276N mutations according to Ambler classification.
17. The method of claim 16 , wherein the subject is being administered or will be administered an antibiotic.
18. 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 A232G, A237S, A238G, S240D, and D276N mutations according to Ambler classification.
19. The method of claim 18 , wherein the subject is being administered or will be administered an antibiotic.