Modified beta-lactamases and methods and uses related thereto
The present invention relates to pharmaceuticals and modified beta-lactamases. Specifically, the invention relates to novel recombinant beta-lactamases and pharmaceutical compositions comprising the beta-lactamases. Also, the present invention relates to methods for modifying a beta-lactamase, producing the beta-lactamase and treating or preventing beta-lactam antibiotic induced adverse effects. Furthermore, the present invention relates to the beta-lactamase for use as a medicament and to the use of the beta-lactamase in the manufacture of a medicament for treating or preventing beta-lactam antibiotics induced adverse effects. Still further, the invention relates to a polynucleotide and a host cell comprising the polynucleotide.
1. A method for reducing the likelihood of a beta-lactam antibiotic-induced adverse effect in a subject in need thereof, comprising administering an effective amount of a beta-lactamase to the subject, wherein:
the subject is receiving a beta-lactam antibiotic, the beta-lactam antibiotic being a substrate of the beta-lactamase,
the beta-lactamase comprises an amino acid sequence having at 90% sequence identity with SEQ ID NO: 1 and an asparagine (N) or arginine (R) residue at a position corresponding to position 276 according to Ambler classification, and
the beta-lactamase hydrolyzes ceftriaxone substantially more efficiently than a beta-lactamase of SEQ ID NO: 1 that has an aspartic acid (D) at a position corresponding to position 276 according to Ambler classification.
2. The method of claim 1 , wherein the beta-lactam antibiotic-induced adverse effect is a Clostridium difficile infection.
3. The method of claim 1 , wherein the beta-lactamase hydrolyses a penicillin and a cephalosporin.
4. The method of claim 3 , wherein the cephalosporin is selected from cefoperazone and ceftriaxone.
5. The method of claim 1 , wherein the beta-lactamase is administered simultaneously to or sequentially with the beta-lactam antibiotic.
6. The method of claim 5 , wherein the beta-lactamase is administered before the beta-lactam antibiotic.
7. The method of claim 1 , wherein the beta-lactamase is administered orally.
8. The method of claim 1 , wherein the beta-lactam antibiotic is administered by intravenous injection.
9. The method of claim 1 , wherein the beta-lactam antibiotic is selected from penicillin and cephalosporin.
10. The method of claim 9 , wherein the cephalosporin is selected from cefoperazone and ceftriaxone.
11. The method of claim 1 , wherein the amino acid sequence has at least 93% sequence identity with SEQ ID NO: 1.
12. The method of claim 1 , wherein the amino acid sequence has at least 95% sequence identity with SEQ ID NO: 1.
13. The method of claim 1 , wherein the beta-lactamase comprises an amino acid sequence of SEQ ID NO: 1 with an asparagine (N) or arginine (R) residue at position 276 according to Ambler classification.
14. A method for reducing the likelihood of a Clostridium difficile infection in a subject in need thereof, comprising administering an effective amount of a beta-lactamase to the subject wherein:
the subject is receiving a beta-lactam antibiotic and the Clostridium difficile infection is caused by unabsorbed beta-lactam antibiotic in the intestinal tract the beta-lactam antibiotic being a substrate of the beta-lactamase,
the beta-lactamase comprises an amino acid sequence having at least 90% sequence identity with SEQ ID NO: 1 and a an asparagine (N) or arginine (R) residue at a position corresponding to position 276 according to Ambler classification and
the beta-lactamase hydrolyzes ceftriaxone substantially more efficiently than a beta-lactamase of SEQ ID NO: 1 that has an aspartic acid (D) at a position corresponding to position 276 according to Ambler classification.
15. The method of claim 14 , wherein the beta-lactamase hydrolyses a penicillin and a cephalosporin.
16. The method of claim 15 , wherein the cephalosporin is selected from cefoperazone and ceftriaxone.
17. The method of claim 14 , wherein the method maintains an ecological balance of normal intestinal microbiota.
18. The method of claim 14 , wherein the amino acid sequence has at least 93% sequence identity with SEQ ID NO: 1.
19. The method of claim 14 , wherein the amino acid sequence has at least 95% sequence identity with SEQ ID NO: 1.
20. The method of claim 14 , wherein the beta-lactamase comprises an amino acid sequence of SEQ ID NO: 1 with an asparagine (N) or arginine (R) residue at position 276 according to Ambler classification.