Antimicrobial and immunostimulatory system comprising an oxidoreductase enzyme

Abstract

The present invention relates to an antimicrobial and immunostimulatory system, applications thereof and a process for the production of the antimicrobial and immunostimulatory system. The present invention provides a storage-stable antimicrobial and immunostimulatory system comprising an oxidoreductase enzyme, a substrate for the oxidoreductase enzyme and hydrogen peroxide in an aqueous solution wherein the substrate for the oxidoreductase enzyme is present up to 90% by weight and water is present up to 20% by weight based on the weight of the total composition; the system has a pH from approximately 4 to 8; and the system provides a two-stage hydrogen peroxide release.

1 .- 58 . (canceled) 
   
   
       59 . A storage-stable antimicrobial and immunostimulatory formulation comprising glucose oxidase, D-glucose, additional sugars selected from one or more of sucrose, fructose and/or maltose and hydrogen peroxide in an aqueous solution;
 wherein glucose oxidase is present at an activity of at least 10 U per 100 g of the formulation;   D-glucose is present from 20 to 85% by weight based on the weight of the total formulation;   additional sugars selected from one or more of sucrose, fructose and/or maltose are present from 5 to 70% by weight based on the weight of the total formulation;   water is present from 10 to 20% by weight based on the weight of the total formulation;   the system has a pH from approximately 4 to 8;   and wherein the formulation provides a two-stage hydrogen peroxide release in which
 storage-stable endogenously produced hydrogen peroxide is bioavailable within the formulation at a level of at least 10 mg per litre for immediate release; and 
 the sustained release of further hydrogen peroxide for at least a twenty-four hour period occurs upon rehydration of the formulation. 
   
   
   
       60 . The formulation of  claim 59  wherein the additional sugars are present from 10 to 70% by weight based on the weight of the total formulation. 
   
   
       61 . The formulation of  claim 59  wherein fructose is present from 8 to 50% w/w %, maltose is present from 4 to 15 w/w %, sucrose is present from 0.5 to 3 w/w % based on the weight of the total formulation. 
   
   
       62 . The formulation of  claim 59  wherein the additional sugars and D-glucose are present at a ratio of approximately 0.05:1 to 3.5:1. 
   
   
       63 . The formulation of  claim 59  wherein the storage-stable endogenously produced hydrogen peroxide is bioavailable within the formulation at a level of at least 75 mg per litre for immediate release. 
   
   
       64 . The formulation of  claim 59  wherein the level of sustained release hydrogen peroxide produced upon rehydration of the formulation is at least 10 mg per litre, preferably 20 mg per litre. 
   
   
       65 . The formulation of  claim 59  further comprising a buffering agent, preferably carbonic acid-bicarbonate and/or phosphoric acid/disodium hydrogen phosphate. 
   
   
       66 . The formulation of  claim 59  further comprising at least one viscosity modifying ingredient. 
   
   
       67 . The formulation of  claim 59  with a pH from approximately 5 to 7. 
   
   
       68 . A storage-stable antimicrobial and immunostimulatory formulation of  claim 59 ,
 wherein the D-glucose is present from approximately 26% to 43% by weight based on the weight of the total formulation;   sucrose is present between 0.5% to 2.5% by weight based on the weight of the total formulation;   fructose is present between 30% to 40% by weight based on the weight of the total formulation;   maltose is present between 5% to 15% by weight based on the weight of the total formulation.   
   
   
       68 a. The formulation of  claim 68 , wherein the maltose is present between 5% and 9%. 
   
   
       69 . The formulation of  claim 68  wherein the D-glucose is present from approximately 33% to approximately 43% by weight based on the weight of the total formulation. 
   
   
       70 . The formulation of  claim 59  in a form adapted for topical, enteral or parenteral administration. 
   
   
       71 . The formulation of  claim 59  in the form of a topical ointment, cream, lotion, oil, liniment, liquid and/or gel, preferably adapted for intramammary administration or adapted for delivery as part of a teat seal, tissue, bandage and/or dressing. 
   
   
       72 . The formulation of  claim 59  in a form adapted for delivery via a dissolvable film strip or strips, dental floss, toothpaste, mouthwash and/or mouth guards. 
   
   
       73 . A method for treating a microbial infection and/or the repair and/or re-growth of damaged tissues and/or cells of a subject comprising the steps of applying an effective amount of the formulation of  claim 59  to an infected area of the subject by topical, enteral and/or parenteral mode of administration. 
   
   
       74 . The method of  claim 73  wherein the microbial infection is caused by gram positive bacteria, gram negative bacteria, acid-fast bacteria, viruses, yeasts, parasitic or pathogenic micro-organisms and/or fungi. 
   
   
       75 . The method of  claim 74  involving the treatment of a wound and/or wound sepsis wherein the wound is an acute wound, chronic wound, surgical wound, chronic burn and/or acute burn. 
   
   
       76 . The method of  claim 73  wherein the microbial infection is an oral, eye and/or ear infection, such as gum disease, oral ulceration and/or an oral hygiene disorder including halitosis and/or gingivitis. 
   
   
       77 . The method of  claim 73  wherein the microbial infection is a skin and/or nail infection, preferably a fungal skin and/or fungal nail infection such as athlete's foot and/or ringworm. 
   
   
       78 . The method of  claim 73  involving the treatment of mastitis, including wet and/or dry mastitis. 
   
   
       79 . A method for teeth whitening wherein the formulation of  claim 59  is in a form adapted for delivery to the teeth of a subject via dissolvable film strip(s), dental floss, toothpaste, mouthwash and/or mouth guard(s) and the method comprises the step of applying the formulation to the teeth of the subject. 
   
   
       80 . A process for manufacture of a storage-stable antimicrobial and immunostimulatory system comprising glucose oxidase, D-glucose, additional sugars selected from sucrose, maltose and/or fructose, and hydrogen peroxide in an aqueous solution according to  claim 59  comprising the steps of
 (a) heating the water to a temperature of at least 60° C., preferably from approximately 75° C. to 95° C.;   (b) adding D-glucose and the additional sugars selected from sucrose, maltose and/or fructose to the heated water to form a water-sugar solution,   (c) cooling the water-sugar solution to a temperature below approximately 40° C. to allow retention of enzyme activity;   (d) adding the glucose oxidase to the water-sugar solution of step (c) with stirring for a sufficient amount of time to form hydrogen peroxide at a pre-determined controlled rate; and   cooling the resultant mixture from step (d) to room temperature to produce a system with bioavailable and storage-stable endogenously produced hydrogen peroxide at a level of at least 10 mg per litre for immediate release.   
   
   
       81 . A storage-stable antimicrobial and immunostimulatory formulation containing glucose oxidase, D-glucose, sucrose, fructose, maltose and hydrogen peroxide in an aqueous solution and an optional buffering agent;
 wherein an effective amount of glucose oxidase is present at an activity of at least 10 U per 100 g of formulation;   wherein the D-glucose is present from approximately 33% to approximately 43% by weight based on the weight of the total formulation;   sucrose is present between 0.5% to 2.5% by weight based on the weight of the total formulation;   fructose is present between 30% to 40% by weight based on the weight of the total formulation;   maltose is present between 5% to 15% by weight based on the weight of the total formulation;   water is present from 10 to 20% by weight based on the weight of the total formulation;   an optional buffering agent in an effective amount of to achieve a formulation with a pH from approximately 4 to 8; and   the formulation provides a two-stage hydrogen peroxide release in which
 storage-stable endogenously produced hydrogen peroxide is bioavailable within the formulation at a level of at least 10 mg per litre for immediate release and 
   the sustained release of further hydrogen peroxide for at least a twenty-four hour period occurs upon rehydration of the formulation.