Lysin polypeptides active against gram-negative bacteria

Abstract

The present disclosure provides methods and compositions useful for the prophylactic and therapeutic amelioration and treatment of infections caused by Gram-negative bacteria, including Pseudomonas aeruginosa. The disclosure further provides compositions and methods of incorporating and utilizing lysin polypeptides of the present disclosure for augmenting the efficacy of antibiotics generally suitable for the treatment of Gram-negative bacterial infection.

1 - 4 . (canceled) 
     
     
         5 . A vector comprising an isolated polynucleotide comprising a nucleic acid molecule that encodes a lysin polypeptide comprising an amino acid sequence having at least 80% sequence identity to a sequence selected from the group consisting of SEQ ID NO: 1-SEQ ID NO: 10, or a fragment thereof having lytic activity. 
     
     
         6 . A recombinant expression vector comprising a nucleic acid encoding a lysin polypeptide comprising an amino acid sequence at least 80% identical to a sequence selected from the group consisting of SEQ ID NO: 1-SEQ ID NO: 10, or a fragment thereof having lytic activity, the nucleic add being operatively linked to a heterologous promoter. 
     
     
         7 . A host cell comprising the vector of  claim 5 . 
     
     
         8 . The vector of  claim 5 , wherein the nucleic acid sequence is a cDNA sequence. 
     
     
         9 . An isolated polynucleotide comprising a nucleic acid molecule that encodes a lysin polypeptide comprising an amino acid sequence having at least 80% sequence identity to a sequence selected from the group consisting of SEQ ID NO: 1-SEQ ID NO: 10, or a fragment thereof having lytic activity. 
     
     
         10 . The polynucleotide of  claim 9  which is cDNA. 
     
     
         11 . A method of inhibiting the growth, or reducing the population, or killing of at least one species of Gram-negative bacteria, the method comprising contacting the bacteria with a composition comprising an effective amount of a lysin polypeptide comprising an amino acid sequence at least 80% identical to a polypeptide sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 4, and SEQ ID NO: 5, or a fragment thereof having lytic activity, wherein the lysin polypeptide or fragment thereof has the property of inhibiting the growth, or reducing the population, or killing  P. aeruginosa  and optionally at least one other species of Gram-negative bacteria. 
     
     
         12 . A method of treating a bacterial infection caused by one or more species of Gram-negative bacteria, comprising administering to a subject diagnosed with, at risk for, or exhibiting symptoms of a bacterial infection, a composition comprising an effective amount of a lysin polypeptide comprising an amino acid sequence at least 80% identical to a polypeptide sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 4, and SEQ ID NO: 5, or a fragment thereof having lytic activity, wherein the lysin polypeptide or fragment thereof has the property of inhibiting the growth, or reducing the population, or killing  P. aeruginosa  and optionally the one or more additional species of Gram-negative bacteria. 
     
     
         13 . The method of  claim 12 , wherein the Gram-negative bacteria is selected from the group consisting of  Pseudomonas aeruginosa, Klebsiella  spp.,  Enterobacter  spp.,  Escherichia coli, Citrobacter freundii, Salmonella typhimurium, Yersinia pestis , and  Franciscella tulerensis.    
     
     
         14 . The method of  claim 12 , wherein the lysin polypeptide amino acid sequence is at least 85% identical to a sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 4, and SEQ ID NO: 5, or a fragment thereof having lytic activity. 
     
     
         15 . The method of  claim 12 , wherein the lysin polypeptide amino acid sequence is at least 95% identical to a sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 4, and SEQ ID NO: 5, or a fragment thereof having lytic activity. 
     
     
         16 . The method of  claim 12 , wherein the Gram-negative bacterial infection is an infection caused by  Pseudomonas aeruginosa.    
     
     
         17 . A method of treating a topical or systemic pathogenic bacterial infection caused by one or more species of Gram-negative bacteria in a subject, comprising administering to a subject a composition comprising an effective amount of a lysin polypeptide comprising an amino acid sequence at least 80% identical to polypeptide sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 4, and SEQ ID NO: 5, or a fragment thereof having lytic activity, wherein the polypeptide or fragment thereof has the property of inhibiting the growth, or reducing the population, or killing the  P. aeruginosa  and optionally the at least one other Gram-negative bacteria. 
     
     
         18 . A method of preventing or treating a bacterial infection comprising co-administering to a subject diagnosed with, at risk for, or exhibiting symptoms of a bacterial infection, a combination of a first effective amount of a composition comprising an effective amount of lysin polypeptide comprising amino acid sequence at least 80% identical to a polypeptide sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 4, and SEQ ID NO: 5, or fragments thereof having lytic activity, and a second effective amount of an antibiotic suitable for the treatment of Gram-negative bacterial infection. 
     
     
         19 . The method of  claim 18 , wherein the antibiotic is selected from one or more of ceftazidime, cefepime, cefoperazone, ceftobiprole, ciprofloxacin, levofloxacin, aminoglycosides, imipenem, meropenem, doripenem, gentamicin, tobramycin, amikacin, piperacillin, ticarcillin, penicillin, rifampicin, polymyxin B, and colistin. 
     
     
         20 . A method for augmenting the efficacy of an antibiotic suitable for the treatment of Gram-negative bacterial infection, comprising co-administering the antibiotic in combination with one or more lysin polypeptides comprising an amino acid sequence at least 80% identical to a polypeptide sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 4, and SEQ ID NO: 5, or a fragment thereof having lytic activity, wherein administration of the combination is more effective in inhibiting the growth, or reducing the population, or killing the Gram-negative bacteria than administration of either the antibiotic or the lysin polypeptide or fragment thereof individually. 
     
     
         21 . The method of  claim 20 , wherein the lysin polypeptide amino acid sequence is at least 90% identical to a sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 4, and SEQ ID NO: 5, or a fragment thereof having lytic activity. 
     
     
         22 . The method of  claim 20 , wherein the lysin polypeptide amino acid sequence is at least 95% identical to a sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 4, and SEQ ID NO: 5, or a fragment thereof having lytic activity. 
     
     
         23 . The method of  claim 20 , wherein the antibiotic is selected from one or more of ceftazidime, cefepime, cefoperazone, ceftobiprole, ciprofloxacin, levofloxacin, aminoglycosides, imipenem, meropenem, doripenem, gentamicin, tobramycin, amikacin, piperacillin, ticarcillin, penicillin, rifampicin, polymyxin B, and colistin. 
     
     
         24 . (canceled)