Beta-lactamases with improved properties for therapy

Abstract

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 - 73 . (canceled) 
     
     
         74 . A beta-lactamase comprising an amino acid sequence having at least 95% sequence identity with SEQ ID NO:1 and having A232G, A237S, A238G, and S240D mutations according to Ambler classification, and further having a mutation at one or more of positions 135, 244, 246, and 276 according to Ambler classification. 
     
     
         75 . The beta-lactamase of  claim 74 , wherein the beta-lactamase has one or more of the following mutations according to Ambler classification:
 a hydrophobic residue other than glutamine (Q) at position 135;   a hydrophobic residue other than arginine (R) at position 244;   a hydrophilic residue other than serine (S) at position 266; and   a hydrophilic residue other than aspartate (D) at position 276.   
     
     
         76 . A polynucleotide comprising a polynucleotide sequence encoding the beta-lactamase of  claim 74 . 
     
     
         77 . A host cell comprising the polynucleotide of  claim 76 . 
     
     
         78 . A pharmaceutical composition, comprising the beta-lactamase of  claim 74  and a pharmaceutically acceptable carrier or excipient. 
     
     
         79 . The pharmaceutical composition of  claim 78 , wherein the pharmaceutical composition is formulated for oral administration. 
     
     
         80 . The pharmaceutical composition of  claim 79 , wherein the pharmaceutical composition is formulated as a tablet, a multi-particulate sprinkle, or a multi-particulate capsule. 
     
     
         81 . 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 having A232G, A237S, A238G, and S240D mutations according to Ambler classification, and further having a mutation at one or more of positions 135, 244, 246, and 276 according to Ambler classification. 
     
     
         82 . The method of  claim 81 , wherein the beta-lactamase has one or more of the following mutations according to Ambler classification:
 a hydrophobic residue other than glutamine (Q) at position 135;   a hydrophobic residue other than arginine (R) at position 244;   a hydrophilic residue other than serine (S) at position 266; and   a hydrophilic residue other than aspartate (D) at position 276.   
     
     
         83 . The method of  claim 81 , wherein the subject is being administered or will be administered an antibiotic. 
     
     
         84 . The method of  claim 81 , 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).