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 95% sequence identity with SEQ ID NO: 1 and the following mutations according to Ambler classification:
an aliphatic hydrophobic residue other than alanine (A) at position 232,
a polar and neutral hydrophilic residue other than alanine (A) at position 237,
an aliphatic hydrophobic residue other than alanine (A) at position 238,
a polar and negative hydrophilic residue other than serine (S) at position 240, and
a polar and neutral hydrophilic residue other than (R) at position 244.
2. The beta-lactamase of claim 1 , wherein the beta-lactamase has a further mutation of a polar and positive hydrophilic residue other than aspartate (D) at position 276 according to Ambler classification.
3. The beta-lactamase of claim 1 , wherein the aliphatic hydrophobic residue is selected from glycine (G), leucine (L), isoleucine (I), methionine (M), and valine (V).
4. The beta-lactamase of claim 1 , wherein the polar and neutral hydrophilic residue is selected from asparagine (N), glutamine (Q), serine (S), threonine (T), proline (P), and cysteine (C).
5. The beta-lactamase of claim 1 , wherein the polar and negative hydrophilic residue is selected from aspartate (D) and glutamate (E).
6. The beta-lactamase of claim 2 , wherein the polar and positive hydrophilic residue is selected from arginine (R) and lysine (K).
7. A polynucleotide comprising a polynucleotide sequence encoding the beta-lactamase of claim 1 .
8. A host cell comprising the polynucleotide of claim 7 .
9. A pharmaceutical composition, comprising the beta-lactamase of claim 1 and a pharmaceutically acceptable carrier or excipient.
10. The pharmaceutical composition of claim 9 , wherein the pharmaceutical composition is formulated for oral administration, optionally selected from a tablet, multi-particulate sprinkle, and a multi-particulate capsule.
11. A method for preventing an antibiotic-induced adverse effect in the gastrointestinal (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 95% sequence identity with SEQ ID NO: 1 and the following mutations according to Ambler classification:
an aliphatic hydrophobic residue other than alanine (A) at position 232,
a polar and neutral hydrophilic residue other than alanine (A) at position 237,
an aliphatic hydrophobic residue other than alanine (A) at position 238,
a polar and negative hydrophilic residue other than serine (S) at position 240, and
a polar and neutral hydrophilic residue other than (R) at position 244.
12. The method of claim 11 , wherein the beta-lactamase has a further mutation of a polar and positive hydrophilic residue other than aspartate (D) at position 276 according to Ambler classification.
13. The method of claim 11 , wherein the aliphatic hydrophobic residue is selected from glycine (G), leucine (L), isoleucine (I), methionine (M), and valine (V).
14. The method of claim 11 , wherein the polar and neutral hydrophilic residue is selected from asparagine (N), glutamine (Q), serine (S), threonine (T), proline (P), and cysteine (C).
15. The method of claim 11 , wherein the polar and negative hydrophilic residue is selected from aspartate (D) and glutamate (E).
16. The method of claim 12 , wherein the polar and positive hydrophilic residue is selected from arginine (R) and lysine (K).
17. The method of claim 11 , wherein the subject is being administered or will be administered an antibiotic.
18. The method of claim 11 , wherein the antibiotic-induced adverse effect in the gastrointestinal (GI) tract is selected from C. difficile infection (CDI), C. difficile -associated disease, and antibiotic-associated diarrhea (AAD).