Therapeutic bacteriophage compositions

Abstract

The present invention provides methods of designing panels of bacteriophages as therapeutic compositions against bacterial infections. The present invention also provides panels of bacteriophages for use in the prevention or treatment of bacterial infections.

1 . A method of designing a panel of bacteriophages as a therapeutic composition against a bacterial infection, the method comprising:
 providing two or more different bacteriophages, wherein each of said two or more different bacteriophages independently retards growth of a target bacterial species or strain;   combining at least two of said two or more different bacteriophages; and   determining growth of the target bacterial species or strain in the presence of said combination of two or more different bacteriophages,   wherein the target bacterial species or strain growth conditions are the same or equivalent in steps (a) and (c);   wherein, if said combination retards growth of the target bacterial species or strain at least equal to the greatest growth retardation achieved independently by any one of said two or more different bacteriophages, the combination is accepted as a panel of bacteriophages; and   wherein, if said combination retards growth of the target bacterial species or strain less than the greatest growth retardation achieved independently by any one of said two or more different bacteriophages, the combination is initially rejected as a panel of bacteriophages.   
     
     
         2 . The method of  claim 1 , further comprising:
 providing at least one further different bacteriophage that independently retards growth of the target bacterial species or strain, and   combining said at least one further different bacteriophage with at least one bacteriophage from a combination specified in  claim 1  parts (d) and (e) to form a further combination; and   determining growth of the target bacterial species or strain in the presence of said further combination;   wherein, if said further combination retards growth of the target bacterial species or strain at least equal to the greatest growth retardation achieved independently by any one of said different or further different bacteriophages, the combination is accepted as a panel of bacteriophages; and   wherein, if said combination retards growth of the target bacterial species or strain less than the greatest growth retardation achieved independently by any one of said different or further different bacteriophages, the combination is further rejected as a panel of bacteriophages.   
     
     
         3 . The method of  claim 2 , wherein, in step (b), said at least one further different bacteriophage is combined with at least one bacteriophage from an initially rejected combination. 
     
     
         4 . The method of any one of  claim 1 , wherein a panel of bacteriophages comprises three or more different bacteriophages. 
     
     
         5 . A method of designing a panel of bacteriophages as a therapeutic composition against a bacterial infection, the method comprising:
 providing a first bacteriophage that retards growth of a target bacterial species or strain;   propagating said first bacteriophage in a culture of the bacterial target species or strain until the development of bacterial resistance against said first bacteriophage, to obtain a first resistant bacterial culture;   providing a second bacteriophage;   determining growth of the first resistant bacterial culture in the presence of said second bacteriophage;   wherein, if said second bacteriophage retards growth of the first resistant bacterial culture, the second bacteriophage is selected for use in a panel of bacteriophages.   
     
     
         6 . The method of  claim 5 , wherein, if said second bacteriophage retards growth of the first resistant bacterial culture, the combination of first and second bacteriophages is selected for use in the panel of bacteriophages. 
     
     
         7 . The method of  claim 5 , further comprising:
 propagating said second bacteriophage in the first resistant bacterial culture until the development of bacterial resistance against said second bacteriophage to form a second resistant bacterial culture;   providing a third bacteriophage;   determining growth of the second resistant bacterial culture in the presence of said third bacteriophage;   wherein, if said third bacteriophage retards growth of the second   resistant bacterial culture, the third bacteriophage is selected for use in the panel of bacteriophages.   
     
     
         8 . The method of  claim 7 , wherein, if said third bacteriophage retards growth of the second resistant bacterial culture, a combination of the third bacteriophage and at least one of the first and second bacteriophages is selected for use in a panel of bacteriophages. 
     
     
         9 . The method of  claim 7 , wherein, if said third bacteriophage retards growth of the second resistant bacterial culture, a combination of the first, second and third bacteriophages is selected for use in a panel of bacteriophages. 
     
     
         10 . The method of  claim 1 , wherein said two or more bacteriophages are provided by first and second bacteriophages as identified in method  claim 5 . 
     
     
         11 . The method of  claim 1 , wherein said two or more bacteriophages are provided by two or more bacteriophages selected from the first, second and third bacteriophages as identified in method  claim 7 . 
     
     
         12 . The method of  claim 5 , wherein a bacterial culture is a bacterial liquid culture. 
     
     
         13 . The method of  claim 1 , wherein growth is determined in a bacterial liquid culture. 
     
     
         14 . The method of  claim 13 , wherein growth is determined in a bacterial liquid culture by measuring optical density of the liquid culture; preferably wherein optical density is measured at 600 nm. 
     
     
         15 . The method of  claim 1 , wherein the bacterial target species or strain is selected from:  Acinetobacter baumanii, Clostridium difficile, Escherichia coli, Klebsiella pneumonia, Pseudomonas aeruginosa, Stenotrophomonas maltophilia , bacterial species causative of body odour,  Staphylococcus aureus , and  Streptococcus mutans.    
     
     
         16 . A bacteriophage panel obtainable by a method according to  claim 1 . 
     
     
         17 . A bacteriophage panel according to  claim 16  for use in the prevention or treatment of a bacterial infection. 
     
     
         18 . The bacteriophage panel for use according to  claim 17 , wherein the subject is a domestic or farm animal. 
     
     
         19 . The bacteriophage panel for use according to  claim 17 , wherein the subject is a human. 
     
     
         20 . The bacteriophage panel for use according to  claim 17 , wherein the bacterial infection is an  Acinetobacter baumanii  infection, a  Clostridium difficile  infection, an  Escherichia coli  infection, a  Klebsiella pneumonia  infection, a  Pseudomonas aeruginosa  infection, a  Stenotrophomonas maltophilia  infection, a  Staphylococcus aureus  infection, or a  Streptococcus mutans  infection. 
     
     
         21 . The bacteriophage panel according to  claim 16  for use in food hygiene. 
     
     
         22 . The bacteriophage panel according to  claim 16  for use in agriculture or crop protection. 
     
     
         23 . The bacteriophage panel according to  claim 16  for use in environmental hygiene applications.