Compositions for controlling bacterial colonization

Abstract

A composition for controlling bacterial growth/colonization is provided. The composition comprises a selected enzyme, a selected anchor molecule coupled to the enzyme to form an enzyme-anchor complex, with the anchor being capable of attaching to a substrate proximal to a bacterial colony. The attachment to the substrate permits prolonged retention time of the enzyme-anchor complex where the bacterial colony is present to increase the effectiveness of the complex. The invention is also for a method of controlling colonization of bacterial plaque in the oral cavity, as well as a method of forming a composition for controlling the proliferation of bacterial colonies in the oral cavity.

1 . A composition for controlling bacterial growth/colonization in the oral cavity comprising: 
 an enzyme,    an anchor molecule coupled to the enzyme to form an enzyme-anchor complex, the anchor being capable of attaching to a substrate proximal to a bacterial colony, the anchor being selected from group consisting of: materials antigenic to substances or surfaces in the oral cavity, bioadhesive molecules, polyhydroxide compounds, salivary components, and oral cavity pellicle components;    wherein the attachment to the substrate permits prolonged retention time of the enzyme-anchor complex where the bacterial colony is present.    
     
     
         2 . A composition as claimed in  claim 1  wherein the materials antigenic to substances or surfaces in the oral cavity are derived from one or more of the group consisting of: saliva; salivary components; pellicle coating in the oral cavity; cellular components; and glycoproteins, glycolipids, proteoglycans and related molecules that contain a sialic acid moiety as part of their constitution.  
     
     
         3 . A composition as claimed in  claim 2  wherein the salivary components are selected from the group consisting of mucopolysaccharides, mucins, glycoproteins, secretary immunoglobulin A, agglutinins, proline-rich proteins (PRPs), histatins, statherin, cystatins, amylases, salivary peroxidases, carbonic anhydrases, lactoferrin, lactoperoxidase, lysozyme, muramidase, kallikrein, and fibronectin.  
     
     
         4 . A composition as claimed in  1  wherein the bioadhesive molecules comprise polypeptides having components selected from at least one of the following: 
 i. rich in basic amino acids;  
 ii. hydroxylated amino acids;  
 iii. DOPA (D or L-3,4-dihydroxyphenyl alanine);  
 iv. copolymers of basic amino acids, DOPA, tyrosine and glutamic acid;  
 V. substances which contain any of the following amino acids which constitute di-, tri-, tetra-, penta-, hexa-, hepta-, octa-, nona- and deca-peptides in any arrangement, combination or sequence: proline, lysine, serine, tyrosine, threonine and alanine; and  
 vi. proline-rich proteins (PRPs).  
 
     
     
         5 . A composition as claimed in  claim 1  wherein the polyhydroxide compounds comprise one or more of molecular units of polyphenols, and polyphenol polymers made up of polyhydroxy or polyphenolic monomeric units.  
     
     
         6 . A composition as claimed in  claim 1  wherein the salivary components and dental pellicles are selected from the group consisting of mucopolysaccharides and mucopolysaccharide fragments, mucins, glycoproteins, histatins, statherin, cystatins, amylases, salivary peroxidases, carbonic anhydrases, lactoferrin, lactoperoxidase, lysozyme, muramidase, kallikrein, fibronectin, and fragments thereof.  
     
     
         7 . A composition as claimed in  claim 1  wherein the interaction between the anchor and the substrate and the anchor and the enzyme is a bonded interaction.  
     
     
         8 . A composition as claimed in  claim 7  wherein the bonded interaction is selected from one or more of: covalent bonds of carbon-to-carbon, carbon-to-nitrogen, sulfur-to-sulfur, cross-linking bonds, ester, ether, peptide, amide and disulfide bonds.  
     
     
         9 . A composition as claimed in  claim 1  wherein the interaction between the anchor and the substrate and the anchor and the enzyme is a non-bonded interaction.  
     
     
         10 . A composition as claimed in  claim 9  wherein the non-bonded interaction is selected from the following interactions: hydrophobic, ionic (charge-charge interactions), receptor-ligand interactions, hydrogen bonding, van der Waals attractive forces.  
     
     
         11 . A composition as claimed in  claim 1  wherein the enzyme is selected for its ability to degrade a colonization matrix.  
     
     
         12 . A composition as claimed in  claim 11  wherein the colonization matrix includes polysaccharides, and the enzyme is selected for its ability to degrade the polysaccharides.  
     
     
         13 . A composition as claimed in  claim 1  wherein the anchor molecule is capable of attaching to any suitable substrate within an oral cavity.  
     
     
         14 . A composition as claimed in  claim 13  wherein the anchor molecule attaches to a tooth surface.  
     
     
         15 . A composition as claimed in  claim 13  wherein the anchor attaches to a pellicle on a tooth surface.  
     
     
         16 . A composition as claimed in  claim 13  wherein the anchor molecule attaches to a bacterial cell wall.  
     
     
         17 . A composition as claimed in  claim 13  wherein the surface in the oral cavity is a plaque matrix.  
     
     
         18 . A composition as claimed in  claim 12  wherein the polysaccharide is glucan.  
     
     
         19 . A composition as claimed in  claim 12  wherein the polysaccharide is a heterogenous and complex aggregate and mixture of many diverse oligo- and polysaccharides.  
     
     
         20 . A composition for controlling bacterial growth/colonization in the oral cavity comprising: 
 an enzyme,    an anchor molecule attachable to the enzyme to form an enzyme-anchor complex, the anchor being capable of attaching to a substrate proximal to a bacterial colony,    an organic molecule having at least two functionalities for binding to the enzyme and the anchor respectively, and    wherein the attachment to the substrate permits prolonged retention time of the enzyme-anchor complex where the bacterial colony is present.    
     
     
         21 . A composition as claimed in  claim 20  wherein the organic molecule is represented by the formula: 
       F z -R-F eb   
       wherein F z  is a charged functionality binding to the anchor, R is an organic moiety and F eb  is the enzyme binding functionality.  
     
     
         22 . A composition as claimed in  claim 21  wherein the organic molecule is a spacer between the anchor and the enzyme.  
     
     
         23 . A composition as claimed in  claim 21  wherein the charged functionality F z  is selected from the group consisting of carboxylate (R—CO 2   − ); sulfonic (R—SO 3   − ); sulfate (R—OSO 3   − ); phosphonic (R—PO 3   − ); phosphate (R—OPO 3   − ); phosphinic (R 1,2 —PO 2   − , where R 1  and R 2  may be identical or dissimilar); any positive species selected from the group including (R x ) 4 N + , ammonium salt R 4 P + , phosphonium salt dipolar functionality including phosphine oxides, phosphoranes, N-oxides, nitro, where there is a charge distribution between the hetero atom and oxygen.  
     
     
         24 . A composition as claimed in  claim 21  wherein R is has one or more of following structures: aliphatic, aromatic, cyclic, heterocyclic, polynuclear aromatic, polymeric (homo- or hetero-monomeric).  
     
     
         25 . A composition as claimed in  claim 21  wherein F z  initially possesses an ionic functionality convertible to a reactive species, so that the initial F z -R-F eb  complex is converted to F zx -R-F eb , where X is a halide atom and the F zx  portion can be secured to a surface or entity within the oral cavity represented by the formula: 
       SURFACE-F z -R-F eb   
       wherein the halide X is released.  
     
     
         26 . A composition as claimed in  claim 25  wherein a sulfonic acid converts to sulfonyl halide.  
     
     
         27 . A composition as; claimed in  claim 20  wherein the organic molecule has multiple functionalities whereby two or more anchors may be attached thereto.  
     
     
         28 . A composition as claimed in  claim 20  wherein the organic molecule has multiple functionalities whereby two or more enzymes may be attached thereto.  
     
     
         29 . A composition for controlling bacterial growth/colonization in the oral cavity comprising: 
 an enzyme selected from the group consisting of a combination of purified enzymes or an ersatz mixture of proteases, lipases and/or glycohydrolases in various ratios;    an anchor molecule coupled to the enzyme to form an enzyme-anchor complex, the anchor being capable of attaching to a substrate proximal to a bacterial colony;    wherein the attachment to the substrate permits prolonged retention time of the enzyme-anchor complex where the bacterial colony is present.    
     
     
         30 . A composition for controlling bacterial growth/colonization in the oral cavity comprising: 
 an enzyme selected from the group consisting of biologic extracts, oxidoreductase enzymes, polysaccharide-degrading enzymes, proteases, carbonic anhydrases, and a mixture of any two or more thereof;    an anchor molecule coupled to the enzyme to form an enzyme-anchor complex, the anchor being capable of attaching to a substrate proximal to a bacterial colony;    wherein the attachment to the substrate permits prolonged retention time of the enzyme-anchor complex where the bacterial colony is present.    
     
     
         31 . A composition as claimed in  claim 30  wherein the biologic extracts are selected from the group consisting of: dehydrated pancreas (Viokase) and combinations of trypsin, chymotrypsin, carboxypeptidase, amylase, lipase, nuclease, and fungal and bacterial extracts containing hydrolytic enzymes.  
     
     
         32 . A composition as claimed in  claim 30  wherein the oxidoreductase is glucose oxidase.  
     
     
         33 . A composition as claimed in  claim 30  wherein the polysaccharide-degrading enzyme is selected from the group consisting of dextranase, mutanase, mucinase, amylase, fructanase (levanase), galactosidase, glucosidase and glucan hydrolases.  
     
     
         34 . A composition as claimed in  claim 30  wherein the protease is neutral, alkaline or acid proteases.  
     
     
         35 . A composition as claimed in  claim 30  wherein the carbonic anhydrases are selected from one or more of metalloenzymes, lysozymes, muramidases, lactoferrin, kallidrein and other serine proteases for the degrading and clearing of debris from the basic polysaccharide backbone structure of plaque.  
     
     
         36 . A method of controlling bacterial colonization comprising the steps of: 
 forming an anchor-enzyme complex comprised of an enzyme selected for its ability to degrade at least portion of a colonization matrix, and an anchor, a portion of which is coupled to the enzyme to produce the complex; and    selecting the anchor based on the ability of said anchor to attach to a substrate, to thereby increase the retention time of the enzyme-anchor complex in close proximity to the matrix.    
     
     
         37 . A method of forming a composition for controlling the proliferation of bacterial colonies, the method comprising: 
 selecting an enzyme based on its ability to degrade the structural component where bacterial colonization occurs;    selecting an anchor molecule based on its ability to couple to the selected enzyme such that the enzyme retains effective enzymatic activity to degrade the structural component, the anchor molecule further being selected for its ability to attach to a substrate proximal the bacterial colonization; and    coupling the anchor and enzyme to produce an enzyme-anchor complex.    
     
     
         38 . A method as claimed in  claim 37  for controlling proliferation of bacterial colonies in the oral cavity.  
     
     
         39 . A method of treatment for controlling the proliferation of bacterial colonies, the method comprising: 
 selecting an enzyme based on its ability to degrade the structural component where bacterial colonization occurs;    selecting an anchor molecule based on its ability to couple to the selected enzyme such that the enzyme retains effective enzymatic activity to degrade the structural component, the anchor molecule further being selected for its ability to attach to a substrate proximal the bacterial colonization;    introducing the anchor molecule in to the oral cavity;    introducing the enzyme in to the oral cavity; and    allowing the anchor and enzyme to couple in the oral cavity to produce an enzyme-anchor complex.    
     
     
         40 . A method as claimed in  claim 39  wherein an intermediate organic molecule having at least two functionalities is attached to the anchor, each functionality binding to an anchor and an enzyme respectively.  
     
     
         41 . A composition as claimed in  claim 1  wherein the anchor is a non-dialyzable material obtained from cranberry juice.  
     
     
         42 . A composition as claimed in  claim 1  wherein the anchor is an adhesin receptor structure.  
     
     
         43 . A composition as claimed in  claim 1  wherein the anchor is an adhesin molecule.  
     
     
         44 . A composition as claimed in  claim 1  wherein the anchor is a bacterial lectin.  
     
     
         45 . A composition as, claimed in  claim 1  wherein the anchor is selected from a glucan-binding lectin (GBL), a glucan-binding protein (GBP), a glucosyltransferase, and a glucan-binding domain (GBD) of a whole or part of a glucosyltransferase enzyme.  
     
     
         46 . A composition as claimed in  claim 1  wherein the anchor attaches to lipoteichoic acid.  
     
     
         47 . A composition as claimed in  claim 1  wherein the anchor is antigenic to lipoteichoic acid.