Compositions and methods for inhibiting biofilm deposition and production

Abstract

The invention provides a method for combating biofilm, said method comprising contacting a biofilm with a composition comprising effective amounts of one or more biofilm degrading enzymes in combination with antimicrobial agents or antimicrobial essential oils. The biofilm may be on an animate or inanimate surface and both medical and non-medical uses and methods are provided. In a preferred aspect the invention provides a composition and method for use in the treatment or prevention of a biofilm infection in a subject, particularly in the oral cavity.

1 . A composition comprising one or more essential oils and at least one biofilm degrading enzyme which act synergistically to degrade biofilm structures and inhibit biofilm deposition, in a biologically acceptable carrier for delivery of said composition. 
     
     
         2 . The composition as claimed in  claim 1  where the essential oils comprise one or more of menthol, thymol, eucalyptol and methyl salicylate and the at least one biofilm degrading enzyme is selected from mutanase, dextranase and glucoamylase. 
     
     
         3 . The composition as claimed in  claim 2  where the essential oils are menthol, thymol, eucalyptol and methyl salicylate, said biofilm degrading enzymes are mutanase and dextranase. 
     
     
         4 . The composition as claimed in  claim 2 , wherein said biofilm degrading enzymes are produced in a plant plastid. 
     
     
         5 . The composition of  claim 3  further comprising glucoamylase. 
     
     
         6 . The composition of  claim 1  wherein said biofilm degrading enzymes are dextranase to mutanase ratio in said composition is 5:1. 
     
     
         7 . A composition comprising an effective amount of one or more biofilm degrading enzymes selected from mutanase, dextranase and glucoamylase and one or more essential oils selected from the group consisting of menthol, thymol, eucalyptol, methyl salicylate, and combinations of two or more thereof in a carrier suitable for oral delivery, wherein said carrier is selected from the group consisting of mouthwash, mouth rinse, mouth spray, toothpaste, chewing gum, tooth gel, sub-gingival gel, mousse, foam, chewable tablet, dentifrice, lozenge, and dissolvable strip. 
     
     
         8 . The composition of  claim 7 , wherein said carrier is mouthwash, mouth rinse, or mouth spray. 
     
     
         9 . The composition of  claim 7 , wherein the carrier is a chewing gum, chewable tablet, lozenge, or dissolvable strip. 
     
     
         10 . The composition of  claim 7 , wherein the carrier is a chewing gum. 
     
     
         11 . The composition of  claim 7 , wherein said essential oils comprise a combination of menthol, thymol, eucalyptol and methyl salicylate. 
     
     
         12 . A method of degrading and/or removing unwanted biofilm from the oral cavity, comprising administering an effective amount of one or more biofilm degrading enzymes selected from the group consisting of mutanase, dextranase, glucoamylase, and combinations of two or more thereof, and one or more essential oils selected from the group consisting of menthol, thymol, eucalyptol, methyl salicylate, and combinations of two or more thereof, to a surface of the oral cavity having said biofilm, said enzymes degrading said film and said essential oil(s) exerting a bactericidal effect, thereby synergistically reducing or eliminating said unwanted biofilm. 
     
     
         13 . The method of  claim 11 , wherein the one or more essential oils comprises at least two essential oils selected from menthol, thymol, eucalyptol and methylsalicylate. 
     
     
         14 . The method of  claim 13 , wherein said essential oils comprise a combination of menthol, thymol, eucalyptol, and methyl salicylate. 
     
     
         15 . The method of  claim 12 , which is effective to selectively kill pathogenic  S. mutans  without adversely affecting commensal bacteria,  A. naeslundii  and  S. oralis.    
     
     
         16 . The method of  claim 12  wherein said administering step comprises first administering the biofilm degrading enzymes followed by administering the essential oils. 
     
     
         17 . A method for inhibiting biofilm deposition on the surface in the oral cavity comprising contacting a surface of the oral cavity susceptible to biofilm deposition with an effective amount of one or more biofilm degrading enzymes selected from the group consisting of mutanase, dextranase, glucoamylase, and combinations of two or more thereof, and one or more essential oils selected from the group consisting of menthol, thymol, eucalyptol, methyl salicylate, and combinations of two or more thereof, to a surface of the oral cavity having said biofilm said enzymes degrading said biofilm and said essential oils exerting a bactericidal effect, thereby synergistically inhibiting deposition of said unwanted biofilm. 
     
     
         18 . The method of  claim 17 , wherein the antimicrobial oral rinse comprises at least two essential oils selected from menthol, thymol, eucalyptol and methylsalicylate. 
     
     
         19 . The method of  claim 18 , wherein said essential oils comprise a combination of menthol, thymol, eucalyptol, and methyl salicylate. 
     
     
         20 . The method of  claim 17 , which is effective to selectively kill pathogenic  S. mutans  without adversely affecting commensal bacteria,  A. naeslundii  and  S. oralis.    
     
     
         21 . The method of  claim 17  wherein said administering step comprises first administering the biofilm degrading enzymes followed by administering the essential oils. 
     
     
         22 . The composition of  claim 4 , further comprising a plant remnant. 
     
     
         23 . The composition of  claim 20 , wherein said plant is a tobacco or lettuce plant. 
     
     
         24 . The composition of  claim 1 , wherein said enzymes are recombinant and are expressed as fusion proteins in bacteria and purified therefrom. 
     
     
         25 . The composition of  claim 1 , wherein said enzymes are purified from non recombinant enzyme expressing bacteria.