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 98% sequence identity with SEQ ID NO: 1 and a mutation at all of positions 232, 237, 238, and 240, according to Ambler classification.
2. The beta-lactamase of claim 1 , wherein the beta-lactamase has a further mutation at position 276 according to Ambler classification.
3. The beta-lactamase of claim 2 , wherein the beta-lactamase has one or more of 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 positive hydrophilic residue or a polar and neutral hydrophilic residue other than aspartate (D) at position 276.
4. A polynucleotide comprising a polynucleotide sequence encoding the beta-lactamase of claim 1 .
5. A host cell comprising the polynucleotide of claim 4 .
6. A pharmaceutical composition, comprising the beta-lactamase of claim 1 and a pharmaceutically acceptable carrier or excipient.
7. The pharmaceutical composition of claim 6 , wherein the pharmaceutical composition is formulated for oral administration, optionally selected from a tablet, multi-particulate sprinkle, and a multi-particulate capsule.
8. 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 98% sequence identity with SEQ ID NO: 1 and a mutation at all of positions 232, 237, 238, and 240, according to Ambler classification.
9. The method of claim 8 , wherein the beta-lactamase has a further mutation at position 276 according to Ambler classification.
10. The method of claim 9 , wherein the beta-lactamase has one or more of 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 positive hydrophilic residue or a polar and neutral hydrophilic residue other than aspartate (D) at position 276.
11. The method of claim 8 , wherein the subject is being administered or will be administered an antibiotic.
12. The method of claim 8 , 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).