JP2018100296A - Antimicrobial compositions - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide antimicrobial compositions.SOLUTION: Storage-stable compositions for generating antimicrobial activity are described. The compositions comprise an enzyme that is able to convert a substrate to release hydrogen peroxide, and an unrefined natural substance, such as a honey, that includes a substrate for the enzyme. In certain embodiments, the enzyme is a purified enzyme. In other embodiments, the substrate lacks catalase activity, and the enzyme is additional to any enzyme activity able to convert the substrate to release hydrogen peroxide that may be present in the unrefined natural substance. The storage-stable compositions do not include sufficient free water to allow the enzyme to convert the substrate. Use of the compositions for treating microbial infections and wounds is described, as well as methods for producing the compositions.SELECTED DRAWING: None

Description

  The present invention relates to compositions for generating antibacterial activity, in particular storage-stable compositions, antibacterial compositions and solutions, the use of these compositions and solutions, in particular for wound healing, and methods for their production.

  Honey has been used since ancient times to treat microbial infections. In recent years, there has been a renewed interest in the therapeutic effect of honey, especially in the field of wound healing. In clinical trials, honey is an effective broad-spectrum antibacterial agent, effective against common wound-infecting microorganisms such as Pseudomonas aeruginosa, Staphylococcus aureus, Candida albicans and Escherichia coli, and against bacterial antibiotic-resistant strains Have been shown to be effective. Honey is also a natural product, making it an attractive alternative to drug-based therapies.

  Many different types of honey have antibacterial activity. This activity is mainly due to osmolality, pH, hydrogen peroxide production and the presence of phytochemical components. Manuka honey produced from the manuka tree (Leptosperum scoparium) has superior antimicrobial activity compared to most other honeys due to the high level of antimicrobial phytochemical activity present in this type of honey. Recognized. However, Manuka Honey has a relatively limited supply and cannot meet global market demand.

  In honey, in addition to its antibacterial action, several other actions are believed to help wound healing, particularly chronic wound healing. In particular, honey has the ability to self-melt wounds and to deodorize wounds. Debridement is the removal of dead, damaged or infected tissue to improve the healing ability of the remaining healthy tissue. Honey has also been reported to have anti-inflammatory properties, be able to stimulate tissue growth, and cope with pain to minimize scarring.

  Several honey-based wound care dressings are currently commercially available. Many of these use manuka honey, or other honey with high levels of non-peroxide antibacterial activity. This is because the antibacterial effect of hydrogen peroxide can be reduced by acidity, catalase activity, and protein digestive enzymes present in wound exudates.

  Honey-based dressings currently on the market include the Medihoney product line containing Manuka honey, manufactured by Comvita / Derma Sciences, and Activon, an additive-free pure Manuka honey manufactured by Advancis. It is done. Other honey-based dressings contain honey and one or more additional ingredients. Examples of these include Meladarm Plus, an antibacterial wound gel manufactured by SanoMed. This gel contains honey from a multi-flowered mountainous region of Bulgaria, which has a naturally high content of glucose oxidase and phenol (the main phytochemical components in honey are phenolic compounds) . During processing, the honey is not heated or irradiated. This is because it is believed that the healing properties, particularly the production of hydrogen peroxide by glucose oxidase, disappear. Instead, sterilization is performed using an ozone treatment method described in WO2008 / 049578. In this method, ozone gas is blown into the liquefied honey in a ozone-resistant container. However, ozone has been approved by the US Food and Drug Administration (FDA) as a disinfectant only for reusable medical devices and is therefore approved by the US FDA for disinfecting honey products for wound healing applications. It has not been.

  The Mesitlan family is a further family of honey-rich wound care products. L-Mesitran ointment manufactured by Triticum is 48% medical grade honey and several other ingredients including lanolin, sunflower oil, cod liver oil, marigold, aloe vera, vitamins C and E, and zinc oxide. Containing.

  The difference in antibacterial activity between honeys can be greater than 100 times depending on the geographical, seasonal and botany sources of honey, as well as harvesting, processing and storage conditions. Therefore, honey-based coating materials have different antibacterial effects depending on the type of honey used. Research by Jenkins, Burton, and Cooper (“The determination of anti-microbiological of the materials that are ti” and “A”. It has been found to have different efficacy against EMRSA-15. The dressings impregnated with Activon were the most effective, followed by the Mediaphone coating, and the effectiveness of the Mesitran product was quite low.

  Therefore, there is a need to reduce the variability in the antibacterial activity of honey and to improve the antibacterial activity of honey with a low antibacterial activity. It would also be desirable to provide an effective honey-based wound care product that has a high level of phytochemical components regardless of the use of honey. It would also be desirable to provide an effective honey wound care product that is not sterilized by ozone treatment.

  Several other antimicrobial wound care dressings are available that do not contain honey. Examples of these include silver-containing dressings such as ConvaTec's Aquacel® Ag dressing, which is a controlled form of silver ions when wound exudate is absorbed into the dressing. To release. However, silver-resistant organisms such as Staphylococcus aureus, Pseudomonas aeruginosa and enterococci have been reported and wound healing may be slowed.

  Povidone iodine (PVP-I) is a stable chemical complex of polyvinylpyrrolidone (povidone, PVP) and simple iodine. This is a broad spectrum disinfectant used topically for wound treatment and infection prevention. It has been demonstrated that bacteria do not develop resistance to PVP-I. PVP-I has been approved by the US FDA since 1994 for first aid treatment of small acute wounds. However, there has been some debate over its safety and efficacy and the US Department of Health and Human Services has not recommended its use in pressure ulcers.

  Chlorhexidine has a rapid bactericidal activity against a wide range of asporic bacteria. Antibacterial activity against Staphylococcus aureus, Pseudomonas aeruginosa and various clinical isolates has been recorded. However, it has been observed that methicillin resistant Staphylococcus aureus (MRSA) is resistant to chlorhexidine. In the UK, the Pharmaceuticals and Medical Products Regulatory Authority (MHRA) has also issued patient safety warnings about the risk of anaphylactic reactions from the use of medical devices and drugs containing chlorhexidine.

  Accordingly, there is also a need to provide antibacterial wound care products with a broad spectrum of activity that are non-toxic and can be used to treat chronic wounds.

International Publication No. 2008/049578

Jenkins RE, Burton NF, Cooper RA, “The determination of antibiotic activity of three honey dresseds by the avalidation test by the RSA”.

  Applicants recognize that the antimicrobial activity of honey depends on the delicate interrelationship between the natural inhibitor and the active agent, and reduce the variability between the honeys of different types and different harvests to reduce the natural honey. The composition which can produce the antibacterial activity of this substance and can improve the antibacterial property of the honey with poor antibacterial activity is developed. Applicants have also found that such compositions have excellent wound healing properties even after sterilization by gamma irradiation exposure. Applicants recognize that these discoveries also have applicability to other natural materials.

  Applicants have found that the antibacterial activity of the above composition can be precisely increased and controlled over a wide range. This makes it possible to provide a composition having antibacterial activity most suitable for the intended use.

  In a broad sense, the present invention provides a composition for generating antimicrobial activity. The composition includes an enzyme capable of converting a substrate to release hydrogen peroxide and a substance comprising a substrate for the enzyme. This enzyme may be present in any of the above substances for any enzyme activity that can convert a substrate and release hydrogen peroxide (referred to herein as “substrate conversion activity”). Additional (ie, added as a result of human intervention). Preferably, the composition is a storage stable composition that does not contain enough free water for the enzyme to convert the substrate.

  According to the present invention, there is provided a storage stable composition for generating antibacterial activity, the composition comprising a purified enzyme capable of converting a substrate to release hydrogen peroxide; and a substrate for the enzyme. The composition does not contain enough free water for the enzyme to convert the substrate.

  According to the present invention, there is also provided a method for producing a storage-stable composition for generating antibacterial activity, which comprises purifying a purified enzyme capable of converting a substrate and releasing hydrogen peroxide against the enzyme. The composition comprises contacting with a substance comprising a substrate and the composition does not contain sufficient free water for the enzyme to convert the substrate.

  In accordance with the present invention, there is further provided a storage stable composition for generating antibacterial activity, the composition comprising an enzyme capable of converting a substrate to release hydrogen peroxide; and a substrate for the enzyme. Containing and without catalase activity; the composition does not contain enough free water for the enzyme to convert the substrate.

  According to the present invention, there is also provided a method for producing a storage-stable composition for generating antibacterial activity, which method comprises converting an enzyme capable of converting a substrate to release hydrogen peroxide, a substrate for the enzyme. And the composition does not contain enough free water for the enzyme to convert the substrate.

  In the presence of sufficient water, the above-mentioned storage-stable composition enzyme can convert the substrate to release hydrogen peroxide. Hydrogen peroxide is known to be effective against a wide variety of different microorganisms. Antimicrobial activity therefore occurs after dilution of the storage-stable composition of the invention.

  According to the present invention, there is also provided a method for producing an antibacterial composition or solution, which method is sufficient for the enzyme to convert the substrate to release hydrogen peroxide by converting the storage stable composition of the present invention. Diluting with fresh water.

According to the present invention, there is further provided an antimicrobial composition or solution, which is the storage stable of the present invention diluted with sufficient water to release hydrogen peroxide by enzymatic conversion of the substrate. Contains a composition.
For example, the present invention provides the following items.
(Item 1)
A composition for generating antibacterial activity that is stable to storage, wherein the composition converts a substrate to release hydrogen peroxide; and an unpurified natural substance comprising a substrate for the enzyme;
And the composition does not contain enough free water for the enzyme to convert the substrate.
(Item 2)
A composition for generating antibacterial activity that is stable in storage, wherein the composition is capable of converting a substrate to release hydrogen peroxide; and an unpurified composition that has no catalase activity and contains a substrate for the enzyme Natural substances;
Wherein the enzyme is additive to any enzyme activity that may be present in the unpurified natural material and that can convert the substrate to release hydrogen peroxide, Wherein the enzyme does not contain enough free water to convert the substrate.
(Item 3)
The composition according to item 1, wherein the unpurified natural substance has no catalase activity.
(Item 4)
4. A composition according to item 2 or 3, wherein the substance has been treated to remove or inactivate catalase activity.
(Item 5)
A composition according to any preceding item, wherein the substance is an unpurified natural sugar substance.
(Item 6)
Item 6. The composition according to Item 5, wherein the sugar substance is honey.
(Item 7)
6. The composition according to item 5, wherein the sugar substance is honey that has been treated to remove catalase activity or inactivated.
(Item 8)
Item 6. The composition according to Item 5, wherein the sugar substance is syrup or converted syrup.
(Item 9)
The composition according to any one of the preceding items, wherein the substance has no enzyme activity capable of converting the substrate to release hydrogen peroxide.
(Item 10)
The composition according to any one of the preceding items, wherein the substance is a pasteurized substance.
(Item 11)
Item 7. The composition according to Item 6, wherein the honey is a non-pasteurized honey, preferably a creamy honey that has not been pasteurized.
(Item 12)
A composition according to any preceding item, wherein the enzyme is an oxidoreductase enzyme.
(Item 13)
13. The composition according to item 12, wherein the oxidoreductase enzyme is glucose oxidase, hexose oxidase, cholesterol oxidase, galactose oxidase, pyranose oxidase, choline oxidase, pyruvate oxidase, glycolate oxidase or amino acid oxidase.
(Item 14)
14. The composition according to item 13, wherein the substrate for the oxidoreductase enzyme is D-glucose, hexose, cholesterol, D-galactose, pyranose, choline, pyruvic acid, glycolic acid or an amino acid.
(Item 15)
14. The composition according to item 13, wherein the oxidoreductase enzyme is glucose oxidase, and the substrate for the oxidoreductase enzyme is D-glucose.
(Item 16)
16. The composition of item 15, wherein the composition comprises 1 unit or more of glucose oxidase per gram of the composition.
(Item 17)
18. A composition according to item 16, wherein the composition comprises 1500 units or less of glucose oxidase per gram of the composition.
(Item 18)
18. A composition according to item 16 or 17, wherein the composition comprises more than 15 units of glucose oxidase per gram of the composition.
(Item 19)
Item 19. The composition according to any of items 16-18, wherein the composition comprises 100 units or more, preferably 100-500 units of glucose oxidase per gram of the composition.
(Item 20)
20. The composition according to any of items 16-19, wherein the composition comprises 500 units or more, preferably 500-1000 units of glucose oxidase per gram of the composition.
(Item 21)
The composition according to any one of the preceding items, wherein the composition is a food standard composition.
(Item 22)
A composition according to any of the preceding items, wherein the composition is a medical grade or medical device grade composition.
(Item 23)
The composition according to any one of the preceding items, wherein each component of the composition is a natural substance.
(Item 24)
A composition according to any preceding item, wherein the composition is in powder form.
(Item 25)
25. An antimicrobial composition comprising a composition according to any of items 1 to 24 diluted with sufficient water to release hydrogen peroxide by conversion of the substrate by the enzyme.
(Item 26)
A composition for generating antibacterial activity, the composition comprising an unpurified natural substance; and a purified enzyme capable of releasing hydrogen peroxide by converting a substrate in the unpurified natural substance. .
(Item 27)
A composition for generating antibacterial activity, wherein the composition comprises an unpurified natural substance having no catalase activity; and an enzyme capable of releasing hydrogen peroxide by converting a substrate in the unpurified natural substance. And wherein the enzyme is additive to any enzyme activity that may be present in the unpurified natural material and that can convert the substrate to release hydrogen peroxide.
(Item 28)
27. The composition according to item 26, wherein the unpurified natural substance has no catalase activity.
(Item 29)
29. The composition according to any one of items 26 to 28, wherein the composition further comprises hydrogen peroxide.
(Item 30)
The composition according to any of the preceding items, wherein the composition is a sterile composition.
(Item 31)
31. The composition according to item 30, wherein the composition is sterilized by exposure to gamma radiation.
(Item 32)
32. An antibacterial solution comprising the composition according to any of items 26 to 31.
(Item 33)
30. A method of sterilizing a composition according to any of items 1 to 29, wherein the method is preferably 10-70 kGy, more preferably 25-70 kGy, most preferably 35-70 kGy of gamma irradiation. Exposing said method.
(Item 34)
A method for producing a composition for generating antibacterial activity having storage stability, wherein said method comprises a purified enzyme capable of converting a substrate to release hydrogen peroxide, and an unpurified natural substance comprising a substrate for said enzyme The method wherein the composition does not contain sufficient free water for the enzyme to convert the substrate.
(Item 35)
A method for producing a composition for generating antibacterial activity having storage stability, wherein the method comprises an enzyme capable of converting a substrate to release hydrogen peroxide, comprising a substrate for the enzyme and having no catalase activity Contacting the raw natural material, wherein the composition does not contain enough free water for the enzyme to convert the substrate.
(Item 36)
35. A method according to item 34, wherein the unpurified natural substance has no catalase activity.
(Item 37)
37. A method according to any of items 34 to 36, wherein the substance is an unpurified natural sugar substance.
(Item 38)
Any substance present in the substance that can treat the substance prior to contacting the substance with the enzyme to convert the substrate and / or convert the substrate to release hydrogen peroxide. 38. The method according to any one of items 34 to 37, wherein the enzyme activity is removed or inactivated.
(Item 39)
39. The method of item 38, further comprising treating the substance prior to contacting the substance with the enzyme to remove or inactivate catalase activity and / or substrate converting enzyme activity present in the substance.
(Item 40)
Treating the substance by heating the substance to inactivate catalase and / or substrate conversion activity and then cooling the substance to a temperature at which the enzyme can maintain activity before contacting the substance with the enzyme. 40. A method according to item 39.
(Item 41)
41. A method according to any of items 34 to 40, wherein the substance is honey.
(Item 42)
38. A method according to item 37, wherein the unpurified natural sugar substance is syrup or converted syrup.
(Item 43)
43. The method of item 42, wherein the method further comprises converting a material comprising sucrose to form the converted syrup.
(Item 44)
44. The method according to any of items 34 to 43, wherein the enzyme is an oxidoreductase enzyme.
(Item 45)
45. A method according to item 44, wherein the oxidoreductase enzyme is glucose oxidase, hexose oxidase, cholesterol oxidase, galactose oxidase, pyranose oxidase, choline oxidase, pyruvate oxidase, glycolate oxidase or amino acid oxidase.
(Item 46)
46. The method of item 45, wherein the substrate for the oxidoreductase enzyme is D-glucose, hexose, cholesterol, D-galactose, pyranose, choline, pyruvic acid, glycolic acid or an amino acid.
(Item 47)
45. The method of item 44, wherein the oxidoreductase enzyme is glucose oxidase and the substrate for the oxidoreductase enzyme is D-glucose.
(Item 48)
48. The method according to any one of items 34 to 47 for producing a food standard composition, wherein the substance and the enzyme are food standards.
(Item 49)
49. The method according to any of items 34 to 48, wherein each component of the composition is a natural substance (item 50).
A method for producing a composition for generating antibacterial activity, the method comprising contacting a purified enzyme capable of converting a substrate and releasing hydrogen peroxide with an unpurified natural substance comprising a substrate for the enzyme. Including said method.
(Item 51)
A method for producing a composition for generating antibacterial activity, wherein the method comprises an enzyme capable of converting a substrate to release hydrogen peroxide, an unpurified natural substance containing a substrate for the enzyme and having no catalase activity Contacting said method.
(Item 52)
51. A method according to item 50, wherein the unpurified natural substance has no catalase activity.
(Item 53)
53. A method according to any of items 34 to 52, wherein the enzyme and the unpurified natural substance are in powder form.
(Item 54)
25. A method for producing an antibacterial solution, wherein the method is a storage-stable antibacterial composition according to any one of items 1 to 24, wherein the enzyme converts the substrate in the sugar substance to produce hydrogen peroxide. Said method comprising diluting with sufficient water to release.
(Item 55)
33. The composition according to any one of items 1 to 31 for use as a pharmaceutical product or the antibacterial solution according to item 32.
(Item 56)
33. The composition according to any one of items 1 to 31 or the antibacterial solution according to item 32, which is used for prevention, treatment or improvement of microbial infection.
(Item 57)
Item 31. Use of the composition according to any one of Items 1 to 31 or the antibacterial solution according to Item 32 in the manufacture of a pharmaceutical product for preventing, treating or improving microbial infection.
(Item 58)
A method for preventing, treating or ameliorating microbial infection, wherein the method requires the prevention, treatment or amelioration of the composition according to any of items 1 to 31 or the antibacterial solution according to item 32. Said method comprising administering to said subject.
(Item 59)
59. The method of item 58, wherein the method comprises topically administering the composition or the solution.
(Item 60)
60. Use or method according to any of items 55-59 for treating a human or animal subject.
(Item 61)
A method of treating a wound, wherein the method comprises administering the composition of any of items 1-31 or the antimicrobial solution of item 32 to a wound site.
(Item 62)
33. The composition according to any one of items 1 to 31 or the antibacterial solution according to item 32 for wound treatment.
(Item 63)
Item 33. Use of the composition according to any one of Items 1 to 31 or the antibacterial solution according to Item 32 in the manufacture of a medicament for wound treatment.
(Item 64)
A method for treating inflammation, wherein the method comprises administering the composition according to any of items 1 to 31 or the antibacterial solution according to item 32 to an inflamed site.
(Item 65)
33. The composition according to any one of items 1 to 31 or the antibacterial solution according to item 32 for the treatment of inflammation.
(Item 66)
Item 33. Use of the composition according to any one of Items 1 to 31 or the antibacterial solution according to Item 32 in the manufacture of a medicament for treating inflammation.
(Item 67)
A method of stimulating tissue growth comprising administering the composition of any of items 1-31 or the antimicrobial solution of item 32 to a site in need of such stimulation. Said method.
(Item 68)
33. The composition according to any one of items 1 to 31 or the antibacterial solution according to item 32 for stimulating tissue growth.
(Item 69)
Item 33. Use of the composition according to any one of Items 1 to 31 or the antibacterial solution according to Item 32 in the manufacture of a medicament for stimulating tissue growth.
(Item 70)
A method of debridement, the method comprising administering the composition of any of items 1-31 or the antimicrobial solution of item 32 to a wound in need of debridement.
(Item 71)
33. The composition according to any one of items 1 to 31 or an antibacterial solution according to item 32 for debridement.
(Item 72)
Use of the composition according to any of items 1 to 31 or the antibacterial solution according to item 32 in the manufacture of a medicament for debridement.
(Item 73)
A method for deodorizing a wound, the method comprising administering a composition according to any of items 1-31 or an antimicrobial solution according to item 32 to a wound in need of deodorization.
(Item 74)
Item 33. The antibacterial solution according to Item 32 or Item 32 for deodorizing wounds.
(Item 75)
Item 33. Use of the composition according to any one of Items 1 to 31 or the antibacterial solution according to Item 32 in the manufacture of a medicament for deodorizing wounds.
(Item 76)
59. The method of item 58, wherein the subject is an insect.
(Item 77)
77. A method according to item 76, wherein the insect is a bee.
(Item 78)
78. A method according to item 77 for prevention, treatment or amelioration of Nosema infection.
(Item 79)
Item 33. A pharmaceutical composition comprising the composition according to any one of items 1 to 31 or the antibacterial solution according to item 32 together with a pharmaceutically acceptable carrier, excipient or diluent.
(Item 80)
80. The pharmaceutical composition according to item 79, wherein the composition is sterile.
(Item 81)
81. The pharmaceutical composition according to item 79 or 80, wherein the pharmaceutical composition is a medical grade or medical device grade pharmaceutical composition.
(Item 82)
A wound dressing comprising a dressing material for covering a wound and the composition according to any one of items 1 to 31 or the antibacterial solution according to item 32.
(Item 83)
33. An antibacterial composition adapted for topical administration to a subject, wherein the antibacterial composition comprises the composition according to any of items 1-31 or the antibacterial solution according to item 32.
(Item 84)
33. An antibacterial composition adapted for oral administration to a subject, wherein the antibacterial composition comprises the composition according to any of items 1-31 or the antibacterial solution according to item 32.
(Item 85)
33. A kit comprising the composition according to any one of items 1 to 31 or the antibacterial solution according to item 32, and instructions for administration of the composition.
(Item 86)
A method for preventing or inhibiting microbial growth, wherein the method prevents or inhibits such growth of the composition of any of items 1-31 or the antimicrobial solution of item 32. Said method comprising using the desired surface or area.
(Item 87)
89. The method of item 86, wherein said method comprises preventing or inhibiting the growth of plant or entomopathogenic fungi using said composition or said solution.
(Item 88)
88. A method according to item 86 or 87, wherein the method comprises using the composition or the solution to prevent or inhibit the growth of Erwinia.
(Item 89)
89. A method according to any of items 86 to 88, wherein the method comprises using the composition or the solution on a plant, fruit or vegetable.
(Item 90)
90. A method according to item 89, wherein the vegetable is potato.
(Item 91)
33. Use of the composition according to any of items 1 to 31 or the antibacterial solution according to item 32 to prevent or inhibit microbial growth.

The results of the well diffusion assay for the antibacterial effect of pasteurized Ulmo honey supplemented with different amounts of glucose oxidase are shown. Figure 2 shows the results of a well diffusion assay for antibacterial effect of a composition of the present invention with different amounts of enzyme compared to the effect of different phenol standards. 2 shows a photograph of the results of a well diffusion assay for the antibacterial effect of pasteurized Tineo honey supplemented with glucose oxidase. Figure 2 shows the results of a well diffusion assay for the antibacterial effect of different compositions of the invention. Figure 3 shows the results of a well diffusion assay for antibacterial effects of compositions of the present invention comprising different enzyme formulations. Figure 3 shows the results of a well diffusion assay for antibacterial effect of a composition of the invention after 60 or 90 days storage. 2 shows the results of a well diffusion assay for the antimicrobial effect of a powder formulation of the composition of the present invention. 2 shows the results of a well diffusion assay for the antibacterial effect of various products in the presence and absence of the compositions of the present invention. 2 shows the results of a well diffusion assay for the antibacterial effect of various products in the presence and absence of the compositions of the present invention. 2 shows the results of a well diffusion assay for the antibacterial effect of various products in the presence and absence of the compositions of the present invention. 2 shows the results of a well diffusion assay for the antibacterial effect of various products in the presence and absence of the compositions of the present invention. Figure 3 shows the results of a well diffusion assay for the antibacterial effect of a sterile honey-based composition of the present invention compared to Manuka UMF25 + honey. The therapeutic effect of the infected footpad using the aseptic honey composition of the present invention is shown. The therapeutic effect of a footpad ulcer using the sterile honey type composition of this invention is shown. The therapeutic effect of a footpad ulcer using the sterile honey type composition of this invention is shown. The therapeutic effect of the foot ulcer using the aseptic honey composition of the present invention is shown. The therapeutic effect of the foot ulcer using the aseptic honey composition of the present invention is shown. The therapeutic effect of the foot ulcer using the aseptic honey composition of the present invention is shown. The therapeutic effect of the foot ulcer using the aseptic honey composition of the present invention is shown. The therapeutic effect of the traumatic leg wound using the sterile honey-type composition of this invention is shown. The therapeutic effect of the infectious leg wound using the sterile honey-type composition of this invention is shown. The therapeutic effect of the foot ulcer using the aseptic honey composition of the present invention is shown. The therapeutic effect of the leg ulcer using the aseptic honey composition of the present invention is shown. The therapeutic effect of the leg ulcer using the aseptic honey composition of the present invention is shown. The therapeutic effect of the leg ulcer using the aseptic honey composition of the present invention is shown. The therapeutic effect of the leg ulcer using the aseptic honey composition of the present invention is shown. The therapeutic effect of the leg ulcer using the aseptic honey composition of the present invention is shown. The therapeutic effect of the leg ulcer using the aseptic honey composition of the present invention is shown. The therapeutic effect of the leg ulcer using the aseptic honey composition of the present invention is shown. The therapeutic effect of the leg ulcer using the aseptic honey composition of the present invention is shown. The therapeutic effect of the leg ulcer using the aseptic honey composition of the present invention is shown. Figure 2 shows the therapeutic effect of diabetic leg ulcers associated with latent osteomyelitis using the sterile honey-based composition of the present invention. The therapeutic effect of the pressure ulcer using the sterile honey-type composition of this invention is shown. The therapeutic effect of the infectious axillary wound using the aseptic honey type composition of this invention is shown. The therapeutic effect of the infection around the entrance of a catheter using the sterile honey-type composition of this invention is shown. The time kill curves of Surgihoney 1 (S1), Surgihoney 3 (S3), and Medihoney (MH) against different test organisms are shown: (a) Staphylococcus aureus; (b) methicillin resistant Staphylococcus Aureus (MRSA); (d) vancomycin-resistant enterococcus (VRE); (e) Pseudomonas aeruginosa; (f) Klebsiella; (g) E. coli; E. coli ESBL; (h) Enterococcus faecalis. The time kill curves of Surgihoney 1 (S1), Surgihoney 3 (S3), and Medihoney (MH) against different test organisms are shown: (a) Staphylococcus aureus; (b) methicillin resistant Staphylococcus Aureus (MRSA); (d) vancomycin-resistant enterococcus (VRE); (e) Pseudomonas aeruginosa; (f) Klebsiella; (g) E. coli; E. coli ESBL; (h) Enterococcus faecalis. The time kill curves of Surgihoney 1 (S1), Surgihoney 3 (S3), and Medihoney (MH) against different test organisms are shown: (a) Staphylococcus aureus; (b) methicillin resistant Staphylococcus Aureus (MRSA); (d) vancomycin-resistant enterococcus (VRE); (e) Pseudomonas aeruginosa; (f) Klebsiella; (g) E. coli; E. coli ESBL; (h) Enterococcus faecalis. Figure 3 shows the results of a plaque assay testing the antiviral activity of S1 and S2 Surgihoney. 2 shows a photograph of the results of using S1 Surgihoney for treating infectious leg ulcers. (A) shows the first day of treatment; (b) shows the fourth day of treatment; and (c) shows the seventh day of treatment. 2 shows a photograph of the results of using S1 Surgihoney for the treatment of pressure ulcers. (A) shows the first day of treatment; and (b) shows the 30th day of treatment. Figure 2 shows different hydrogen peroxide production rates for Surgihoney (SH) and two modified prototypes, PT1 and PT2. Figure 2 shows the relationship between phenol activity and maximum hydrogen peroxide activity in modified honey SH, PT1 and PT2.

  Catalase is an enzyme that catalyzes the decomposition of hydrogen peroxide into water and oxygen. Using a substance that does not have catalase activity means that there is no variation in the amount of this activity between similar substances from different sources, or between similar substances from different collections from the same source. This reduces the variation in antibacterial activity that can occur from such materials. Alternatively, if the substance contains catalase activity and it is impossible or undesirable to deactivate the catalase activity in the substance before contacting the substance with the enzyme, peroxidation that can be generated from the substance Enough enzyme may be used so that the effect of catalase activity on hydrogen is reduced. This also reduces the variation in antibacterial activity that can be generated from the above substances. In some embodiments, it is preferred that the substance is free of catalase activity.

  Catalase is present in many plants and animals. Catalase activity may be removed during processing or harvesting of the above materials, or may be inactivated prior to use of this material in the compositions of the present invention. Catalase activity may be heat-inactivated by pasteurization, for example. The temperature suitable for heat inactivation of the catalase activity is at least 60 ° C., 70 ° C. or 80 ° C., preferably 2 minutes or longer.

  As used herein, the term “storage-stable” means that the composition is kept at room temperature for several days or more, preferably one week or more, while maintaining the ability to generate antibacterial activity after dilution of the composition. More preferably, it is used to mean that it can be stored for 1 or 2 months or more. The preferred storage temperature is less than 37 ° C, preferably 20-25 ° C. The composition is preferably stored without exposure to light.

  Hydrogen peroxide is generally unstable at room temperature. Insufficient free water in the storage stable composition of the present invention prevents the enzyme from converting the substrate to release hydrogen peroxide, thus making the composition stable at room temperature for extended periods of time. Help maintain. The storage-stable composition of the present invention may contain some water provided that there is not enough free water for the above enzyme to convert the substrate. The amount of suitable water will vary depending on the detailed components of the composition. Typically, however, the shelf-stable composition of the present invention preferably comprises a total water content of less than 20%, for example 10% to 19% water.

  Hydrogen peroxide may be released for a sustained period after dilution of the composition of the invention, depending on the amount of substrate present in the composition and the activity of the enzymes described above. The amount of substrate and / or enzyme activity in the composition is such that, after dilution of the composition, a relatively high level of hydrogen peroxide release in a short time, or a lower level of peroxidation in a longer time. It will be appreciated that one may choose to provide hydrogen release. Preferably, the composition provides a sustained release of hydrogen peroxide for at least 24 hours, more preferably at least 48 hours after dilution of the composition. Preferably, the composition provides a sustained release of hydrogen peroxide at a level of less than 2 mmol / liter after dilution of the composition for at least 24 hours.

  The enzyme present in the composition of the invention may be present in any of the above substances, any enzyme activity capable of converting a substrate to release hydrogen peroxide (herein referred to as “substrate conversion activity”). In other words, the composition of the present invention comprises the above substances and the added enzyme. In some embodiments, it is preferred that the substance does not have substrate conversion activity.

  It is understood that after dilution of the shelf stable composition of the present invention, sufficient enzyme should be present in the composition to convert the above substrate to form hydrogen peroxide as necessary. Like.

  Because it is important that hydrogen peroxide is generated by the storage-stable composition of the present invention in the presence of sufficient water, the composition of the present invention should not contain any added peroxidase. Will be understood.

  Preferably the enzyme is a purified enzyme. As used herein, the term “purified enzyme” is used to include an enzyme preparation in which the enzyme is separated from at least some of the impurities originally present when the enzyme is made. Preferably, impurities that have been removed or reduced include impurities that would otherwise interfere with the enzyme's ability to convert the substrate and release hydrogen peroxide.

  If the enzyme is capable of converting the substrate and releasing hydrogen peroxide, it may not be necessary or desirable that the purified enzyme be of a high level of purity. May be. In some situations, it may be desirable to use an enzyme formulation that is relatively crude. Examples of suitable purity levels include at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80% or 90% purity.

  However, it is preferred that the amount of any catalase that may be present from the outset when the above enzyme is made is reduced. This enzyme may be produced by recombinant means or non-recombinant means, and may be a recombinant enzyme or a non-recombinant enzyme. This enzyme may be purified from a microbial source, preferably from a non-genetically modified microorganism.

  The purity level of the enzyme may be selected as needed depending on the intended use of the composition. For example, if the composition is intended for human consumption, food grade purity may be appropriate. Medical applications require the use of medical grade or medical device grade purity.

  Preferably the enzyme is an oxidoreductase enzyme. Examples of oxidoreductase enzymes that can convert substrate to release hydrogen peroxide include glucose oxidase, hexose oxidase, cholesterol oxidase, galactose oxidase, pyranose oxidase, choline oxidase, pyruvate oxidase, glycolate oxidase and amino acid oxidase Is mentioned. The substrates corresponding to these oxidoreductase enzymes are D-glucose, hexose, cholesterol, D-galactose, pyranose, choline, pyruvate, glycolic acid and amino acids, respectively.

  A mixture of one or more oxidoreductase enzymes and one or more substrates for the oxidoreductase enzyme may be present in the composition of the present invention.

  According to a preferred embodiment of the present invention, the oxidoreductase enzyme is glucose oxidase and the substrate is D-glucose.

  The above substance may be any substance containing a substrate for the above enzyme. Preferably this substance is free of catalase activity. Preferably this material is an unpurified material. As used herein, the term “unpurified” is used to refer to material that has not been processed to a pure form. Unpurified material includes material that may be concentrated, for example, by drying or boiling.

  Preferably, the material includes one or more substrates from natural sources (referred to herein as “natural materials”). Examples of natural substances include substances from plant sources, including substances from sap, roots, nectar, flowers, seeds, fruits, leaves or shoots. More preferably, this material is an unpurified natural material.

  Preferably said substance comprises one or more of the following substrates: D-glucose, hexose, cholesterol, D-galactose, pyranose, choline, pyruvic acid, glycolic acid or amino acid.

Preferably the substance is a sugar substance. As used herein, the term “sugar substance” is used to mean any substance that contains one or more sugars. As used herein, the term “sugar” is used to refer to a carbohydrate having the general formula C _m (H ₂ O) _n . Preferred sugars include monosaccharides such as D-glucose, hexose or D-galactose. Preferably, the sugar material comprises one or more sugars from natural sources (referred to herein as “natural sugar materials”). More preferably, the natural sugar material is an unpurified natural sugar material. The unpurified natural sugar material may be a natural sugar product (or may be derived from a natural sugar product). In a preferred embodiment, the unpurified natural sugar product is honey. In some preferred embodiments, the honey is a honey that has been treated to remove or inactivate catalase activity. Alternatively, the unpurified natural sugar material may be a processed natural sugar such as syrup or inverted syrup.

  As explained above, preferably the substance itself may not have enzyme activity (referred to as “substrate conversion activity”) that can convert the substrate and release hydrogen peroxide. The lack of substrate conversion activity of this substance has the advantage that there is no variation in the amount of this activity between similar substances from different sources, or between similar substances from different sources from the same source. Have. This further reduces the variation in antibacterial activity that can occur from such materials. If so, substrate conversion activity is provided only by the enzyme in contact with the substance, and the amount of substrate conversion activity present in the composition can be controlled.

  Substrate conversion activity may be removed during processing or collection of the above materials, or may be inactivated prior to use of this material in the compositions of the present invention. Substrate conversion activity may be inactivated by heat inactivation, for example by pasteurization. A suitable temperature for heat inactivation of the substrate conversion activity is at least 80 ° C., preferably for at least 2 minutes. The advantage of heat inactivation is that both catalase activity and substrate conversion activity can be inactivated in a single heat inactivation step.

  The storage-stable composition of the present invention may contain an antimicrobial agent. For example, the enzyme is contacted with the substance in an aqueous solution under the conditions of substrate conversion by the enzyme, and then the composition is dried to provide free water that is not sufficient for the enzyme to convert the substrate. Hydrogen peroxide may be present if the above shelf-stable composition is formed by reducing the water content of the composition to a level that only exists. Preferably, however, the storage-stable antimicrobial composition does not contain detectable hydrogen peroxide. Such a composition may be formed, for example, by contacting the enzyme with a substrate in the absence of sufficient free water for the enzyme to convert the substrate. Examples of other antimicrobial agents that may be present in the shelf stable compositions of the present invention include antibiotics, antiviral agents or antifungal agents.

  The composition of the present invention is particularly preferably a food standard composition. Such a composition may be used for human consumption. In other particularly preferred embodiments, the composition is a medical grade or medical device grade composition. Alternatively, the composition of the present invention may be an animal food standard for consumption in animals.

  It is particularly preferred that each component of the composition is a natural substance (ie, each component is derived from a natural source). The compositions of the present invention containing only natural ingredients provide an attractive alternative to drug-based antimicrobial formulations.

  Preferably the substance is honey. The honey may preferably be medical grade or medical equipment grade honey. In some embodiments, the honey is preferably honey that has been treated to remove or inactivate catalase activity originally present in the honey. According to a preferred embodiment of the present invention, the substance is pasteurized honey and the enzyme is glucose oxidase. According to a preferred embodiment, the substance is a medical grade or medical equipment grade honey and the enzyme is a medical grade or medical equipment grade enzyme, preferably glucose oxidase.

  Honey is a natural product made by bees using nectar from flowers. Honey is a saturated or supersaturated solution of sugar. Honey is a natural sweet substance made by bees from the nectar of plants, secretions from live parts of plants, or insect effluents that suck plants at live parts of the plant in the Codex Alimentarius International Food Standards. Honey collected, mixed and mixed with special substances of bees, dehydrated, stored in beehives, left to mature and mature " Revised Coded Standard for Honey, 2001).

  Nectar typically contains about 14% monosaccharide (w / w), 1% phenolic compound, and 85% water. This phenolic compound imparts flavor, aroma and color to the honey. The nectar ferments very fast under the warm conditions of the beehive, typically 36 ° C. To prevent this, nectar is mixed with secretions from the salivary glands and hypopharyngeal glands of foraging bees that contain enzymes. In the beehive, nectar is passed from bees to bees and more secretions are added before they are stored in the beehives. The amount of enzyme present depends on the age of the bees, diet and physiology (when the bees are foraging bees, they produce more digestive enzymes in the glands), colony strength, beehive temperature, and nectar flow And sugar content.

Enzymes added to nectar by bees include diastase, which catalyzes the conversion of starch into dextrin and sugar, invertase, which catalyzes the conversion of sucrose into fructose and glucose, and glucose into hydrogen peroxide and gluconic acid. Glucose oxidase that catalyzes Low doses of hydrogen peroxide prevent the growth of yeast that ferments nectar quickly. When the bees progressively dry the nectar to form honey, the honey becomes acidic (pH 3.5 to 4.5) by gluconic acid. In honey, water is effectively trapped in sugar molecules and cannot be used for further chemical reactions. The amount of “free” water in the honey is measured as water activity (a _w ). The range of a _w found in honey is reported to be 0.47 to 0.70, with mean values of 0.562 and 0.589 (RCIEGG, M; BLANC, B, 1981, The water activity of honey). and related sugar solutions.Lebensmittel-Wissenshaft und Technology 14: 1-6). The _aw of aged honey is so low that no species growth is maintained and fermentation does not occur if the moisture content is lower than 17.1% (Molan, P. et al.
C. (1992). The antibacterial activity of honey: The nature of the antibacterial activity. Bee World, 73 (1), 5-28). The acidity of honey and the absence of free water prevents further fermentability and stops the action of glucose oxidase. Honey also contains an indefinite amount of catalase originating from nectar.

The typical chemical composition of flower honey is as follows.

  In addition, there are traces of pollen and the enzymes invertase, diastase, catalase and glucose oxidase. Pollen can be used to identify the plant origin of honey. There are also phytochemical components. This will vary depending on the source of the honey, but is typically less than about 1%.

  When diluted, glucose oxidase present in natural honey converts the glucose substrate in the diluted honey to release hydrogen peroxide. However, variability in honey content (especially variability in the content of glucose oxidase activity, glucose and catalase activity) can be observed in honey from different sources, or another collection of honey from the same source. This means that the antibacterial efficacy can vary greatly.

  According to an embodiment of the present invention, the honey may be pasteurized. Honey pasteurization inactivates the catalase and glucose oxidase activities present in this honey. Optionally, the pasteurized honey may be filtered to remove any particles (such as wax particles and bee wings) that may be present in the honey after harvest. In order to form the storage stable composition of the present invention, the temperature of the added glucose oxidase is not inactivated, and this pasteurized honey is sufficient to facilitate mixing with glucose oxidase. Once the honey has been cooled to a temperature that remains liquid (preferably 35-40 ° C.), the honey and glucose oxidase are contacted.

  Honey can be pasteurized at a temperature sufficient for heat inactivation of catalase activity. The preferred minimum temperature is 60 ° C to 80 ° C. This temperature should preferably be maintained for at least 2 minutes.

  Since the by-product of heating the honey is the production of HMF (hydroxymethylfurfuraldehyde) used as an indicator of heat and storage changes in the honey, control of the heat treatment may be important. HMF is produced by the decomposition of fructose in the presence of acid. Heat increases the rate of this reaction. The speed increases exponentially with increasing heat. Each time honey rises 1 ° C. above 40 ° C., which is close to the ambient temperature of a normal honeycomb, HMF increases rapidly. HMF is not a hazardous product. Jam, molasses, golden syrup, etc. may have an HMF concentration 10 to 100 times that of honey. However, HMF concentration is used as an indicator of honey degradation, and according to Codex Alimentarius Standard, in the EU, 40 mg / l is the maximum allowable concentration in honey for a table.

  To prevent an increase in HMF, the honey is rapidly raised to a temperature level at which catalase is inactivated, and then the temperature of the honey is rapidly increased up to 40-45 ° C. using a heat exchange mechanism. It is preferable to lower.

  No water is added during the process of this preferred embodiment, so the resulting composition does not contain enough free water to convert glucose in the presence of glucose oxidase to release hydrogen peroxide. The above shelf-stable composition includes pasteurized honey and added glucose oxidase. There is no detectable hydrogen peroxide. This composition can be stored at room temperature for several days or longer.

  In other embodiments of the invention, the honey may not be pasteurized.

  According to some preferred embodiments, the honey (pasteurized or non-pasteurized) is a creamy honey. Creamy honey is honey that has been processed to control crystallization. Creamy honey contains a large number of small crystals that prevent the formation of large crystals that can occur in raw honey. A process for making creamy honey was described in US Pat. No. 1,987,893. In this processing, the raw honey is first pasteurized, and then a pre-processed cream honey is added to the pasteurized honey to produce a mixture of 10% cream honey and 90% pasteurized honey. The mixture is then rested at a temperature controlled at 14 ° C. This method produces a batch of creamy honey in about a week. Seed batches can be made by crystallizing normal honey and crushing the crystals to the desired size. Large scale manufacturers modified the process by stirring the mixture using a stir bar while keeping the honey mixture at 14 ° C. In an alternative creaming method, the pasteurization step may be omitted, and instead the honey is slowly warmed to 37 ° C.

  The glucose oxidase is preferably a purified natural glucose oxidase formulation, which is a food standard for human consumption or of medical grade or medical equipment grade. The activity of the glucose oxidase may be selected according to a desired generation rate of hydrogen peroxide generated after dilution of the storage-stable composition. Several glucose oxidase formulations are commercially available (glucose oxidase is identified by reference CAS: 9001-37-0). Common microbial sources of glucose oxidase from non-genetically modified organisms include selected strains of Aspergillus niger, Penicillium amagasakiense, Penicillium variabile, and Penicillium notatum. Medical device grade glucose oxidase from GMO Aspergillus niger is available from Biozyme UK and has an activity of 240 iu / mg. Food grade glucose oxidase from Aspergillus niger is available from BIO-CAT INC and has an activity of 15,000 units / g. Non-genetically modified glucose oxidase is available from BIO-CAT INC and has an activity of 12,000 / g. Glucose oxidase (GO3B2) from Aspergillus niger is available from BBI Enzymes Limited and the activity is 360 units / mg. Contaminants: alpha amylase 0.05% or less, saccharase 0.05% or less, maltase 0.05% or less, and GO / Cat2000 or more.

  The activity (eg, glucose oxidase activity) of the above enzyme may be in the range of, for example, 1 to 400 IU / mg or 1 to 300 IU / mg, for example 250 to 280 IU / mg. The amount of enzyme used depends on the desired use of the composition, the amount of any catalase activity present in the material, the amount of substrate present in the material, the desired level of hydrogen peroxide release, and the desired excess. This may be due to several factors, including the length of hydrogen oxide release time. Suitable amounts of enzyme can be readily determined by one skilled in the art and, if necessary, hydrogen peroxide for different amounts of enzyme using a well diffusion assay as described in Example 2 below. The extent of release can be measured and determined. A suitable amount of enzyme (such as glucose oxidase) may be 0.0001% to 0.5% w / w of the above composition. The amount of enzyme used may be selected to produce a composition that generates antibacterial activity equal to the selected phenol standard (eg, 10%, 20% or 30% phenol standard).

  The composition of the present invention, particularly the above-mentioned substance is honey (for example, non-pasteurized honey), and the above enzyme can convert D-glucose in this honey to release hydrogen peroxide. The composition of the present invention that is glucose oxidase may contain 1 unit or more, preferably 1500 units or less of glucose oxidase per gram of the composition. This glucose oxidase is additive (ie, added as a result of human intervention) to any glucose oxidase that may be naturally present in the material.

  As used herein, “unit” is defined as the amount of enzyme that oxidizes 1 micromole of glucose per minute at 25 degrees Celsius and pH 7.0.

  Applicants have found that the antibacterial activity of the compositions of the present invention may be increased simply by increasing the amount of glucose oxidase activity present in the composition.

In some embodiments of the invention, the composition of the invention is greater than 15 units per gram of the composition, such as at least 30 units, at least 50 units or at least 100 units, and preferably less than 685 units, such as Contains 100-500 units of glucose oxidase. Such compositions have been found to have superior antibacterial properties than compositions having 15 units or less of glucose oxidase per gram of the composition. In particular, such compositions are disclosed in MSSA, MRSA, Group A and B Streptococci, Enterococcus, E.I. E. coli, E.I. coli
ESBL, Serr. lifacfaciens Amp C, Kleb. pneumoniae, Pseud. Has high potency against a wide range of microorganisms including aeruginosa and Candida albicans.

  In other embodiments of the invention, the composition of the invention comprises at least 500 units, such as 500-1000 units or 685-1000 units of glucose oxidase per gram of the composition. Such compositions have been found to have even better antimicrobial properties. In particular, such compositions are MSSA, MRSA, E.I. It has higher potency against a wide range of microorganisms including E. coli ESBL and Staphylococcus aureus.

  The pasteurization process described above inactivates any enzyme activity present in the honey, thus catalase activity between pasteurized honey from different sources or between different collections of honey from the same source. And no variation in substrate conversion activity. The amount of substrate exchange activity can be controlled by the addition of a purified glucose oxidase formulation with a well-defined amount and a well-defined enzyme. Thus, the inherent antimicrobial activity variability between honeys of different types and different harvests is significantly reduced, and the antimicrobial properties of honey with low antimicrobial activity are improved.

  According to a further preferred embodiment of the invention, said substance may be a conversion substance, preferably a conversion syrup such as a converted maple syrup. As used herein, the term “inverted” is used to mean that sucrose initially present in the substance has been converted to glucose and fructose, for example, by the activity of an invertase or sucrase enzyme. Accordingly, such additive materials may be used in the compositions of the present invention having a glucose oxidase enzyme.

  Maple syrup is made from the sap of sugar maple or black maple trees. In cold climates, these trees store starch on their stems and roots before winter, and when the starch is converted to sugar, it rises into the sap in the spring. Maple trees can be pierced and drained, and the exuded sap can be collected and concentrated to produce maple syrup. Maple syrup typically contains about 66% sucrose, 33% water, and 1% glucose and fructose. Because of the low amount of glucose present, maple syrup does not have strong antimicrobial development properties when mixed with glucose oxidase. However, when sucrose is first converted to glucose and fructose, for example by treatment with invertase or sucrase, the resulting converted syrup contains a much higher concentration of substrate for glucose oxidase.

  According to the present invention, there is further provided an antibacterial composition or a composition for generating antibacterial activity comprising a substance having no catalase activity and an enzyme capable of converting a substrate in the substance and releasing hydrogen peroxide. The

  According to the present invention, an antibacterial composition or for generating antibacterial activity, comprising contacting an enzyme capable of converting a substrate to release hydrogen peroxide with a substance containing a substrate for the enzyme and having no catalase activity. A method of making the composition is also provided.

  According to the present invention, there is further provided an antibacterial composition or a composition for generating antibacterial activity comprising a substance and a purified enzyme capable of converting a substrate in the substance to release hydrogen peroxide.

  According to the present invention, the manufacture of an antibacterial composition or a composition for generating antibacterial activity comprising contacting a purified enzyme capable of converting a substrate to release hydrogen peroxide with a substance containing a substrate for the enzyme. A method is also provided.

  The composition or antimicrobial composition of the present invention may further comprise hydrogen peroxide. According to the present invention, there is further provided an antibacterial solution comprising the composition or antibacterial composition of the present invention.

  The compositions and solutions of the present invention can be used to treat any microbial infection that can be treated with hydrogen peroxide. Examples include infections caused by gram positive bacteria, gram negative bacteria, acid-fast bacteria, viruses, yeasts, parasitic or pathogenic microorganisms or fungi. In particular, infections caused by the following microorganisms may be treated. Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, Candida albicans, Propionibacterium acnes, Staphylococcus aureus, Staphylococcus epidermidis, Staphylococcus saprophytics, beta-hemolytic Streptococci Group A or B, Campylobacter coli, Campylobacter jejuni, methicillin-resistant Staphylococcus Aureus (MRSA), methicillin-sensitive Staphylococcus Aureus (MSSA), Botrytis cinerea, M cobacterium tuberculosis, Cryptosporidium, Plasmodium and Toxoplasma.

  According to the present invention there is also provided the use of a composition or solution of the present invention that prevents or inhibits microbial growth.

  According to the present invention, there is also provided a composition or solution of the present invention for use as a medicament. According to the present invention, there is further provided a composition or solution of the present invention for the prevention, treatment or amelioration of microbial infection. The invention also provides the use of a composition or solution of the invention in the manufacture of a medicament for the prevention, treatment or amelioration of a microbial infection.

  According to the present invention, there is further provided a method for preventing, treating or ameliorating a microbial infection, which comprises administering a composition or solution of the present invention to a subject in need of such prevention, treatment or amelioration. including. The subject may be a human or animal subject. The composition of the present invention may be administered topically.

  Applicants have found that the compositions of the present invention, in particular the compositions of the present invention in which the above-mentioned unpurified natural substance is honey, have excellent wound healing properties. In particular, chronic wounds lasting from one month to over a year are beginning to heal visibly within a few days of treatment with the composition of the present invention.

  In particular, the wound healing activity of the compositions as described above is believed to be due to the sustained, low concentration of hydrogen peroxide release that results when the compositions of the present invention are diluted, for example, with wound exudate. This will carry oxygen to the wound site.

  Oxygen has a variety of important roles in assisting wound healing. Complex wound healing processes require large amounts of energy. If the wound is infected, more energy is needed, which means that more oxygen is needed. Oxygen is involved in many mechanisms of the natural healing process. Oxygen has a major role in metabolic assistance, substrate repair, antiseptic / infection control, and signaling and control of cellular responses. Wounds that receive sufficient oxygen generally heal at a greater rate compared to wounds that do not have enough oxygen. Ischemia / hypoxia directly inhibits wound healing processes such as angiogenesis, collagen synthesis and epithelialization, and may also interfere with the ability of leukocytes to kill bacteria. As the bacteria grow, more white blood cells are recruited to the wound site and oxygen consumption is further increased.

  In order for a wound to heal, it must have sufficient energy and nutrition to drive the healing process. Oxygen is essential for these metabolic processes. Tissue regeneration and healing requires granulation tissue formation, epithelialization, contraction and remodeling. The healing process requires the growth of various types of cells as the matrix forms, new blood vessels form, and the epithelium replaces. Such cellular activity depends on oxygen availability, which allows breathing unimpeded. The biosynthetic pathway required to make all biopolymers (eg proteoglycans, structural proteins, etc.) depends on oxygen, which satisfies the basic energy requirements. Various enzymes are required, including matrix metalloproteases, which are also very expensive in terms of energy demand, ie oxygen demand.

  Collagen synthesis is an important part of the above process. Collagen deposition is very important for repair of connective tissue and as part of the angiogenic process. Oxygen is a cofactor required for the hydroxylation of proline and lysine during the formation of procollagen. The synthesis of mature collagen requires prolyl hydroxylase and lysyl hydroxylase enzymes, both of which depend on oxygen for their function.

  Angiogenesis / angiogenesis is essential to complete healing. This needs to be triggered by an appropriate signal. In a hyperoxic environment, macrophage leukocytes trigger this signal leading to a complex and organized series of events, including collagen formation and organization after tissue degradation, endothelial cell migration / colony formation, and angiogenesis be able to.

When a wound is created, the body's natural defenses are activated. Neutrophils gather at the wound site immediately after trauma and release bactericidal reactive oxygen species (ROS) and hydrogen peroxide (H ₂ O ₂ ) to kill bacteria and prevent infection. In response to environmental stimuli, macrophages arrive at the wound site, engulf foreign bodies, and release vascular endothelial growth factor (VEGF), a very important angiogenic factor for wound healing. Oxygen has a major role in these events. One of the major microbicidal mechanisms of macrophages and neutrophil leukocytes is the “respiration burst”, which is the natural activity by which these cells kill microorganisms. Individuals who are deficient in the respiratory burst process suffer from bacterial infections. Leukocytes require oxygen to carry the respiratory burst effect, and their efficacy can be increased by high oxygen concentrations. These leukocytes also have other bactericidal mechanisms that are triggered when leukocytes engulf microorganisms (phagocytosis). This involves a significant amount of energy consumption and is therefore an oxygen dependent process and therefore does not work well in an oxygen deficient environment. Works best in oxygen-rich conditions. Oxygen also kills anaerobic bacteria and is important for some antibiotics to function effectively.

  Oxygen molecules are important cellular signals that interact with growth factors and other signals (eg, redox signals) to control signal transduction pathways. The molecular signal released by macrophages and triggering angiogenesis is “vascular endothelial growth factor” (VEGF). When the oxygen concentration is high, macrophage leukocytes can release VEGF. Several other important factors and genes associated with enzymes are triggered by high oxygen concentrations.

  Nitric oxide (NO) production is also recognized as central signaling and controls events in wound healing. NO is made by nitric oxide synthase (NOS) enzyme, and its inducible enzyme (iNOS) is upregulated in the early stages of wound healing. However, the enzyme can only function if it is rich in arginine and oxygen, allowing NO production at an appropriate rate.

  Oxygen also has a direct role in signaling (stimulation) of the epithelialization process. The epithelialization process is a major late stage healing event in which new epithelial cells proliferate, organize themselves and differentiate into structured epithelia.

  Therefore, the continuous supply of oxygen to the wound site with the composition of the present invention is considered particularly important in promoting wound healing.

  In addition to the hydrogen peroxide releasing activity of this composition, several activities of this composition are believed to contribute to the wound healing activity of the preferred compositions of the present invention. In particular, preferred compositions have the ability to debride and deodorize wounds, have anti-inflammatory properties, can stimulate tissue growth, and cope with pain to minimize scarring It is considered possible.

  Malodor is a common feature of chronic wounds and is attributed to the presence of anaerobic bacterial species that produce malodorous compounds from spoiled serum and tissue proteins. In addition to the antibacterial action, the composition of the present invention containing glucose (preferably 24 to 40% by weight) metabolizes it to bacteria in preference to amino acids, resulting in the production of odorless metabolite lactic acid.

(Preferably, range close to the honey, that is a with a _w of 0.47 to 0.7) having a high osmolality compositions of the present invention, when promoting debriding by autolysis action of tissue proteases considered It is done. The composition of the present invention creates a moist wound environment by extracting lymph from wound tissue due to its strong osmotic action. This provides a constant supply of protease at the interface between the wound bed and the necrotic tissue overlying the wound bed. This action also cleans the wound bed surface from below. Activation of the protease by hydrogen peroxide released by the composition of the invention in contact with water may also help. The debridement may also contribute to reducing the bacterial load of the wound by removing necrotic tissue. It is well known that necrotic tissue provides a good medium for bacterial growth and increases the risk of infection when left in a wound.

  Although the body's inflammatory response is characteristic of the initiation of the healing process, prolonged reactions can inhibit healing, cause further damage to tissues and make it more difficult to deal with wounds. Prolonged inflammatory responses are often accompanied by high levels of exudate. Inhibiting inflammation not only reduces patient pain, but also reduces vascular opening, thus reducing edema and exudate. The ability of the composition of the present invention to remove infection and debride is believed to contribute to anti-inflammatory effects. Compositions of the invention comprising an antioxidant (preferably one or more antioxidants present in honey) may also reduce excessive inflammation by removing free radicals.

  Prolonged inflammation results in fibrosis that appears as a hypertrophic scar in the wound. Preferred compositions of the invention may reduce scarring by reducing inflammation, promoting angiogenesis, and stimulating granulation tissue formation. Preferred compositions of the invention may also stimulate epithelial growth.

  The compositions of the present invention are also believed to have an immunostimulatory effect mediated by interleukin-1 (IL-1). The compositions of the present invention are believed to promote the release of IL-1 from skin cells. IL-1 is a cytokine that is also secreted by macrophages, monocytes and dendritic cells. IL-1 is an important part of the body's inflammatory response to infection. IL-1 increases adhesion factor expression in endothelial cells, allowing leukocytes to migrate to the site of infection. It also works in the brain's thermoregulatory center to raise body temperature. Called intrinsic pyrogen. As body temperature rises, the body's immune system helps fight infection. This is the early stage of the inflammatory immune response that increases the antimicrobial activity of the immune system. The inflammatory response plays a central role in wound healing through defense against possible infections and by participating in cell and tissue repair and regrowth. The antibacterial effect of the compositions of the present invention is believed to be aided and supplemented by an immunostimulatory effect that aids in regrowth and repair of damaged tissue and / or cells.

  The composition of the present invention, particularly the honey-based composition, provides a moist healing environment for the wound tissue without the risk of maceration of the surrounding skin and prevents the dressing from adhering to the wound bed, thus replacing the dressing There is no pain and no tissue damage. Such compositions also stimulate normal cell growth and accelerate the granulation and epithelialization process in the wound. This composition also has an occlusive effect to protect wounds and damaged tissue. Honey-based compositions are also believed to significantly reduce the pH of the wound and aid the healing process.

  According to a preferred embodiment of the present invention, the composition of the present invention may be used in a method of wound care, including the treatment of wounds or the treatment or treatment of wound sepsis.

  The wound may be an acute wound, a chronic wound, a surgical wound (eg, a cesarean wound), a chronic burn or an acute burn. The compositions of the present invention may be used in the prophylactic prevention of wound sepsis. It will be appreciated that when using the shelf stable composition of the present invention, it may be diluted with the liquid present at the wound site, thereby leading to the release of hydrogen peroxide from the diluted composition. .

  When any tissue integrity is compromised, wounds occur (eg, skin cracks, muscle tears, burns or fractures). A wound may be caused by an act (trauma) or a surgical procedure, by an infection, or by an underlying condition.

  Acute wounds include surgical wounds and trauma such as lacerations, abrasions, tears, penetrations or bites, and burns. Acute wounds typically go through an orderly and timely recovery process, resulting in sustained repair of anatomical and functional integrity.

  Chronic wounds exist for more than 3 weeks or do not go through an orderly and timely process that results in anatomical and functional integrity, or do not build sustained and functional results, A wound that has not undergone the repair process (Lazarus et al., Arch Dermatol. 1994; 130 (4): 489-493).

  Chronic wounds can be divided into four categories: venous ulcers, arterial ulcers, diabetic ulcers and pressure ulcers. The few wounds that do not fall into these categories may be due to reasons such as radiotoxicity or ischemia.

  Venous ulcers usually occur in the legs, occupy about 70% to 90% of chronic wounds, and often occur in the elderly. They are thought to be due to venous hypertension caused by the failure of the valves present in the veins to prevent blood regurgitation. Ischemia results from dysfunction and, when combined with reperfusion injury, causes tissue damage leading to the wound. In venous disease, ulcers are usually in the gait area between the ankle and calf, often on the medial side of the leg.

  Arterial leg ulcers result from decreased arterial blood flow and subsequent tissue perfusion. Atherosclerosis or peripheral vascular disease is the most common cause of arterial leg ulceration. If the reduced blood supply is left untreated, tissue necrosis can occur to the extent that is supplied by the affected artery. Ulcer growth is often rapid, resulting in deep tissue destruction. Arterial leg ulcers can occur anywhere in the lower leg. Often they are deep and circular with well defined boundaries.

  Diabetic ulcers: Diabetes causes immune depression and damages microvessels, preventing proper oxygen supply to tissues, which can lead to chronic wounds. Diabetes causes neuropathy, which impairs pain sensation and pain perception. Thus, the patient may initially be unaware of small wounds on the legs and feet and as a result may not be able to prevent infection or repeated injury. Pressure also plays a role in the formation of diabetic ulcers. Diabetic patients have a 15% higher risk of amputation due to chronic ulcers than the general population.

  Pressure sores usually occur in people with symptoms such as paralysis, which prevents the movement of well-pressed body parts such as the heel, scapula and sacrum. Pressure ulcers are caused by ischemia that occurs when the pressure at the tissue is greater than the pressure at the capillaries, thus restricting blood flow to that area. Muscle tissue requires more oxygen and nutrients than the skin and is most affected by long-term pressure. Like other chronic ulcers, reperfusion injury damages tissue.

  When treating ulcers, creating a barrier and protecting the wound during healing; providing a moist wound environment; removing the wound scab; reducing bacterial load; ideally immune modification and nutrition improvement It is desirable to use to promote healing. Conventional wound treatment products are unable to achieve all of these effects. Some conventional treatments have some of these effects, but they are quite toxic and are therefore not ideal treatments. However, the composition of the present invention, particularly the composition of the present invention in which the above substance is honey, has all these properties. Furthermore, the antibacterial activity of the compositions of the present invention can be adjusted to be much more effective than other honey wound care products, and the composition includes antibiotic resistant bacteria such as MRSA, It is effective against a wide range of pathogenic bacteria related to chronic wounds.

  The compositions of the invention are particularly advantageous in the initial treatment of wounds, particularly in patients such as diabetics whose wounds are likely to get worse. Initial treatment with the compositions of the present invention prevents complications arising from chronic wounds, keeps patients active and prevents consumption in dealing with complications.

  It has also been found that the compositions of the present invention are effective in reducing or preventing surgical wound infections. Surgical wound infection is particularly a problem with cesarean section, with cesarean section having a fairly high infection rate of about 10%. The composition of the present invention is convenient for application after surgery. For example, the composition of the present invention may be applied to a dressing and then contacted with a wound and held in place with a second dressing. Example 40 below demonstrates that a single application of the composition of the present invention to a post-operative cesarean wound reduced the surgical site infection rate by 60% compared to past data.

  Peripherally inserted central venous catheters (PICC lines) are used for chemotherapy treatment and / or administration of other drugs. The PICC line is a long thin flexible tube that is inserted into one of the vena cava in the arm near the elbow bend. It is then passed through the blood vessel until the tip enters the vena cava just above the heart. However, infection can occur within the line or in an area that enters the blood vessel. If infection occurs, patients are usually given antibiotics. If these do not remove the infection or if the infection is severe, the line may be removed. In order to reduce the chance of infection, a dressing containing an antibacterial agent such as chlorhexidine or silver may be used at the site where the line enters.

  The composition of the present invention has been found to be effective in preventing and eliminating colonization in the PICC line by using the composition of the present invention locally at the site where the line enters (see below). See Example 39).

  The compositions of the present invention can be used easily without any expert training or complex equipment. This composition is very suitable for use in the third world. In the third world, patients are often more susceptible to infection, and the materials and equipment used for surgery and wound care are more often infected than in developed countries.

  The compositions of the present invention are non-toxic and do not have any of the problems associated with silver-containing coatings, PVP-I or chlorhexidine.

  The compositions of the present invention may also be used to treat donor or recipient transplant sites.

  In accordance with the present invention, a method of treating a wound is provided, which method comprises administering a composition of the present invention to the wound.

  According to the present invention, there is also provided a composition of the present invention for wound treatment.

  According to the present invention, there is further provided the use of the composition of the present invention in the manufacture of a medicament for the treatment of wounds.

  According to the present invention, a method for treating inflammation is also provided, which method comprises administering a composition of the present invention to the site of inflammation.

  According to the present invention, there is also provided a composition of the present invention for treating inflammation.

  The present invention further provides the use of the composition of the present invention in the manufacture of a medicament for treating inflammation.

  According to the present invention, a method for stimulating tissue growth is also provided, which method comprises administering a composition of the present invention to a site in need of such stimulation.

  According to the present invention there is also provided a composition of the present invention for stimulation of tissue growth.

  According to the present invention, there is further provided the use of the composition of the present invention in the manufacture of a medicament for stimulating tissue growth.

  According to the present invention, a method of debridement is also provided, which method comprises administering a composition of the present invention to a wound in need of debridement.

  According to the present invention, a composition of the present invention for debridement is also provided.

  The present invention further provides the use of the composition of the present invention in the manufacture of a medicament for debridement.

  According to the present invention, a method for deodorizing a wound is also provided, which method comprises administering a composition of the present invention to a wound in need of deodorization.

  According to the present invention there is also provided a composition of the present invention for wound deodorization.

  According to the present invention, there is further provided the use of the composition of the present invention in the manufacture of a medicament for deodorizing wounds.

  For wound healing applications, the compositions of the invention may be administered at an appropriate frequency as determined by the health care provider. Suitably, the composition of the invention may be administered at least every 2-3 days, eg weekly, but preferably every day or every other day.

  The amount of the composition of the invention to be administered depends on many factors such as the strength of the antimicrobial properties of the composition and other wound healing properties of the composition, the size of the wound, and the age and symptoms of the subject being treated. Depends on. For many applications, however, administration of 2 to 100 g or 5 to 100 g of the composition of the present invention may be suitable, preferably 10 to 50 g.

  According to a preferred embodiment of the present invention, the composition of the present invention is sterile. Sterile compositions are preferably used in medical applications such as wound healing.

  The composition of the present invention may be sterilized by any suitable means. Applicants have found that the compositions of the present invention retain glucose oxidase activity (and therefore the ability to release hydrogen peroxide by dilution) after sterilization by exposure to gamma radiation.

  Thus, according to the present invention, there is provided a composition of the present invention that is sterilized by exposure to gamma radiation.

  According to the present invention, there is also provided a method for sterilizing the composition of the present invention, which method comprises exposing the composition to gamma irradiation.

  A suitable gamma irradiation level is 10 to 70 kGy, preferably 25 to 70 kGy, more preferably 35 to 70 kGy.

  Since ozone is not approved by the US FDA for sterilization of honey-based products for wound healing applications, the compositions of the present invention are preferably not sterilized by ozone treatment, ozone or by ozone treatment. It does not contain any ingredients that have been subjected to sterilization. In particular, the composition of the present invention should not contain ozonized honey or ozonated oil.

  A preferred composition of the present invention for medical use is a sterile disposable composition.

  A sterile composition of the present invention stored without exposure to light is expected to remain stable for more than 6 months. For example, such compositions may be packaged in high density polyethylene / low density polyethylene (HDPE / LDPE) tubes or polyester-aluminum-polyethylene (PET / Al / PE) sachets.

  The composition of the present invention is preferably a medical grade or medical device grade composition. Preferably, the unpurified natural substance is honey, preferably medical grade or medical equipment grade honey.

  Preferably, the composition of the present invention comprises creamed honey, more preferably cream honey that has not been pasteurized. Such compositions can be easily administered topically because the large crystals or many large crystals present are minimized by the creaming process.

  It will be appreciated that in compositions of the invention that include honey, it is not necessary to use pasteurized honey in the composition if the composition is sterilized. Alternatively, it may be preferable to use non-pasteurized honey (preferably creamy honey) or other unpurified natural substances. In a particularly preferred embodiment, the composition of the present invention comprises non-pasteurized honey and added purified glucose oxidase.

  Therefore, according to the present invention, a storage-stable composition for generating antibacterial activity is provided, which composition comprises honey that has not been pasteurized and added purified glucose oxidase, the glucose oxidase being sufficient In the presence of free water, D-glucose in honey can be converted to release hydrogen peroxide, and the composition contains sufficient free water for the glucose oxidase to convert D-glucose. Absent.

  According to the present invention, there is also provided a method for producing a storage-stable composition for generating antibacterial activity, the method comprising contacting non-pasteurized honey with purified glucose oxidase, the glucose oxidase Can convert D-glucose in honey to release hydrogen peroxide in the presence of sufficient free water, and the composition contains enough free water for the glucose oxidase to convert D-glucose. Not included.

  Such compositions preferably contain at least 1 unit and preferably 1500 units or less of glucose oxidase per gram of the composition. Suitably such a composition comprises more than 15 units of glucose oxidase per gram of the composition, for example at least 100 units, preferably 100-500 units of glucose oxidase per gram of the composition, or 1 gram of the composition. Per unit of at least 500 units, preferably 500-1000 units of glucose oxidase.

  The honey of such a composition may include cream honey that has not been pasteurized.

  The present invention also provides a pharmaceutical composition comprising the composition of the present invention together with a pharmaceutically acceptable carrier, excipient or diluent.

  According to certain preferred embodiments of the present invention, a dressing comprising the composition of the present invention is provided. Suitable dressings include gauze, bandages, tissue paper, films, gels, foams, hydrocolloids, alginate, hydrogels, or polysaccharide pastes, polysaccharide granules or polysaccharide beads. The compositions of the present invention may be present with a wound dressing substrate, such as a collagen substrate or a collagen-glycosaminoglycan substrate.

  The composition of the present invention may be in the form of a solid or semi-solid formulation. Examples of solid or semi-solid formulations include capsules, pellets, gel capsules, powders, hydrogels, pills, pills or spheres. Alternatively, the composition of the present invention may be in the form of a liquid formulation. Examples of liquid formulations include syrup, paste, spray, drop, ointment, cream, lotion, oil, liniment or gel. Typical gels include alcoholic gels such as isopropanol gel, ethanol gel or propanol gel, or hydrogels.

  The composition of the present invention may be in a form suitable for administration to a human or animal subject. Suitable forms include forms adapted for topical or oral administration. Forms suitable for topical administration include topical ointments, creams, lotions, oils, liniments, liquids, gels or soluble strips. Forms suitable for oral administration include capsules, pellets, gel capsules, pills, small pills, spheres, troches, dental floss, toothpaste, mouthwash, and soluble film strips. When using the storage-stable composition of the present invention, it is diluted with the liquid present at the site of administration (for example, by saliva for oral administration or by exudate from the wound) and peroxidized at the site of administration. Hydrogen may be released.

  The compositions of the present invention may be one or more suitable antibacterial or immunostimulatory ingredients, excipients or adjuvants, or any other suitable where it is desired to provide the ability to generate antibacterial activity. It may be present with the ingredients. For example, the compositions of the present invention may be present with preservatives, antitussives, diaper care formulations, moisturizers, itching mitigants, cleansers, scrubs, detergents, barrier agents, release agents. Preferably, however, the composition of the invention does not contain any antibiotics.

  The compositions of the present invention may be in a form suitable for controlled release or sustained release delivery. For example, oral dosage forms may have enteric coatings to provide controlled release or sustained release delivery.

  According to another aspect of the present invention, the composition of the present invention may be in the form of a use as a cosmetic composition. The compositions of the present invention may be present with one or more suitable cosmetic excipients or adjuvants. Cosmetic applications include many different personal care applications, including treatment of hair conditions such as dandruff or treatment of body odor.

  The composition of the present invention may be provided in the form of a prophylactic hand barrier solution or a hand sanitizing solution. Such hand barrier liquids may be provided in the form of creams, lotions or hydrogels and are used as hand-wash detergent type products with advantageous properties for preventing microbial infections prophylactically.

  Advantageously, the compositions of the invention may be used in the treatment or prevention of MRSA or other antibiotic resistant microorganisms and bacteria. The present invention overcomes the problem of the emergence of microbial antibiotic-resistant strains by non-toxic means. For this application, the compositions of the invention may be administered topically as, for example, topical ointments, creams, lotions, oils, liniments, liquids and / or gels. The compositions of the present invention may be administered as part of a tissue paper or skin wipe.

  Said microbial infection may be an infection of the mouth, eyes, ears, skin, chest or nails. Oral infections may be gingival diseases, oral ulcers and / or oral hygiene disorders. This oral health disorder may be bad breath and / or gingivitis. Alternatively, the oral infection may be a nasal infection, including a pharyngeal infection, or a nasal Staphylococci infection. Ocular infections may include conjunctivitis. The skin infection may be a skin fungal infection. Cutaneous fungal infections include foot ringworm and / or ringworm in humans. In veterinary medicine, cutaneous fungal symptoms include control of hoofs, ringworm and zoonotic skin infections. The nail infection may be a fungal nail infection such as onychomycosis.

  The composition of the present invention may be used for the treatment of skin disorders such as acne, eczema or psoriasis. Acne and eczema may have a microbial infection component that can be treated by this composition, and secondary microbial infections of psoriatic lesions caused by scratching can be treated by the composition of the present invention.

  The composition of the present invention may be used in veterinary medicine. Important veterinary applications include the treatment of microbial infections and the treatment or management of wound care, or the treatment of burns. Specific symptoms include chronic skin infections in dogs (subcutaneous staphylococcus infection), otitis externa (ear infections), oral care in animals, Campylobacter infection in chickens, scoliosis, enteric microbial infections in pigs, poultry and cattle Cryptosporidium infection, elimination of communicable communicable infections, wound dressings such as horn removal and abscess treatment. The present invention has the particular advantage that in veterinary applications it becomes possible to treat microbial infections without the inclusion of antibiotics in the food chain.

  The compositions of the present invention may be provided with any other composition or product where it is desired to provide the ability to generate antimicrobial activity. The compositions of the invention may be provided as a mixture with other compositions or products, or may be provided separately from them, eg packaged as a kit with other compositions or products. When the composition of the present invention is provided as a mixture with other compositions or products, the other compositions or products are free enough for the enzyme described above to convert the substrate of the composition of the present invention. It is preferable not to contain water.

  The composition of the present invention may be provided with instructions for using the composition. For example, the composition of the present invention may be packaged as a kit with the instructions.

  According to the present invention, there is provided a kit for prevention, treatment or amelioration of microbial infection, or wound treatment, which kit comprises the composition or solution of the present invention and instructions for administration of the composition or solution. Including.

  In accordance with the present invention, there is further provided a method for preventing or inhibiting microbial growth, wherein the method uses the composition or solution of the present invention on a surface or area where it is desired to prevent or inhibit microbial growth. including.

  The compositions or solutions of the present invention may be particularly useful for preventing or inhibiting the growth of plant or insect pathogens. For example, the compositions or solutions of the present invention can prevent shelf life of edible products (such as fruits or vegetables) or other products (expensive plant material including bulbs, especially lily bulbs, or flowers), or prevent microbial growth. It may be particularly suitable to extend by blocking. If this microbial growth is not prevented or inhibited, the product will be damaged. Treatment of edible products or other products with the compositions or solutions of the present invention may be preferable to using pesticides or pharmaceutical products, especially when the compositions or solutions contain only natural ingredients. Good.

According to a preferred embodiment, the composition or solution of the present invention may be used to prevent or inhibit the growth of Erwinia. Erwinia is a genus of the family Enterobacteriaceae and mainly contains plant pathogen species. These species produce enzymes that hydrolyze pectin between plant cells, leaving the cells apart. This is a disease called “plant rot”. Well-known members of this genus are E. Amylovora species, which cause burn disease in apples, pears and other rosaceae crops. Erwinia carotovora (Pectobacte
rium carotovorum), another species, has a broad host range including carrots, potatoes, tomatoes, leafy vegetables, pumpkins and other pumpkins, onions, peppers. This species can cause disease in almost any plant tissue that it invades. This is a very important pathogen because it is a common cause of spoilage in preserved fruits and vegetables. E. Rot caused by carotovora is often referred to as bacterial soft rot (BSR).

  Erwinia infection is particularly problematic for potato preservation. Significant losses can occur when Erwinia occurs. It also causes black rot at the center of the potato. This problem is currently controlled using pharmaceutical products. According to a preferred embodiment of the present invention, the composition or solution of the present invention may be used for the prevention or prevention of potato rot caused by Erwinia infection. It is particularly preferred to use a composition or solution according to the invention which contains only natural ingredients.

  According to a further aspect of the present invention, the composition or solution of the present invention may be used to prevent or inhibit the growth of insect pathogens, especially Nosema in honey bees.

  Nosema is the most prevalent adult bee disease. This is caused by a unicellular microsporozoan parasite that exists in two stages, the long-lived spore stage and the replication-proliferation stage. When an adult honeybee ingests a spore, the spore grows to the growth stage and invades cells that form the inside of the bee's intestine. Two species that affect bees, Nosema apis and Nosema ceranae, have been described. Nosema apis reduces the life span of infected bees. Nosema ceranae has transcended species barriers from Asian hosts. It is known to be more pathogenic than apis. Infected foraging honeybees die away from the beehive, thus losing honeybees continuously, reducing food delivery to the beehive and then destroying the colony.

  Treatment of Nosema with the antibiotic Fumidil B (prepared from Aspergillus fumigatus, causative agent of Stonebrood disease) inhibits spore replication in the midgut but does not kill the spores. This antibiotic is also very expensive. It must also be used off-season to minimize the risk of contaminating market honey and entering the human food chain.

  We have found that the compositions of the present invention can kill Nosema spores and Nosema at the growth stage. According to the present invention, there is provided a method for preventing or treating Nosema in a bee, the method comprising administering to the bee a composition or solution of the present invention. Preferably, the storage-stable composition of the present invention in powder form is diluted with water and used in beehives. The storage-stable composition of the present invention in powder form may be made, for example, by dry pasteurizing honey prior to contacting with glucose oxidase in powder form.

  Preferred embodiments of the present invention will now be described, by way of example only, with reference to the accompanying drawings.

Example 1
This example describes a preferred manufacturing process for the storage-stable composition of the present invention and a preferred process for diluting the storage-stable composition to release hydrogen peroxide.

“Activated” Honey Manufacturing Process Heat the honey for 2 minutes at 80 ° C. using a heat exchanger (if desired, use a lower temperature if sufficient to inactivate catalase) Preferably 60 ° C. or higher). The purpose of this heating process is to pasteurize the honey and to reduce the honey viscosity so that it can be filtered to remove any wax particles and wings that may be present in the honey after collection. As well as inactivating any catalase in honey that affects the effective generation of hydrogen peroxide.

  The pasteurized honey is then filtered and left to cool naturally to the normal honeycomb temperature (35-40 ° C.). Glucose oxidase is then added at a low concentration (equal to that normally found in honey) to replace the glucose oxidase naturally present in the honey but inactivated by the pasteurization process. The filtered pasteurized honey is fairly liquid at 35-40 ° C. and can be easily mixed with the glucose oxidase. However, there is no reason that this mixing cannot be performed at room temperature. The resulting “activated” honey is then stored at room temperature.

  There is no modification to the natural honey composition other than the replacement of the inactivated glucose oxidase. There is no detectable hydrogen peroxide in the above “activated” honey.

Dilution of “activated” honey After dilution of “activated” honey, free water becomes available and the glucose oxidase starts to convert the glucose present in this honey, so that over-oxidation occurs over time. Hydrogen is released. The hydrogen peroxide concentration is less than 2 mmol / liter but is released for a long time.

Example 2
This example describes the results of a test demonstrating the antibacterial effect of activated Ulmo honey. This honey is described as “activated” if it contains added glucose oxidase.

Well Diffusion Assay Staphylococcus aureus (NCIMB 9518) was grown on nutrient agar or in nutrient broth.

  The culture was seeded on an antibacterial diffusion agar plate by collecting the overnight culture with a cotton swab and placing it on the surface of the agar plate. The plate was left at room temperature for 15 minutes. A well with a diameter of 7 mm was opened on the agar surface. 200 microliters of sample (phenol standard or honey) was placed in each well.

  Plates were incubated for 16 hours and inhibition circles were measured using dial calipers (+/− 0.1 mm). The diameter of the zone including the well diameter was recorded.

  A phenol standard was prepared by diluting phenol in purified water at the required concentration. For example, a 10% phenol standard was prepared by diluting 1 g of phenol in 9 g of purified water. This standard was stored at 30 ° C. and shaken before use.

Honey Honey: Pasteurized Ulmo Honey

  Non-activated honey (ie, honey without added glucose oxidase) was used as a control.

Enzyme preparation Glucose oxidase: medical device grade material, non-food grade, GMO Aspergillus niger origin, supplied by Biozyme UK, activity 240 iu / mg. Initially 0.5% w / w enzyme was used, but this could be reduced to 0.005% w / w enzyme to equal the 20% phenol standard.

Hydrogen peroxide detection Merckquant peroxide test: catalog number 1.10011.0002 Measuring range / color scale memory mg / l H2O2 0.5-2-5-10-25.

Measurement procedure in aqueous solution: Dissolve honey in water at 50/50 w / w. The reaction zone of the test strip is immersed in the measurement sample (15-30 ° C.) for 1 second. Run excess liquid from the long end of the test strip onto the absorbent paper towel and after 15 seconds (Catalog No. 110011) or 5 seconds (Catalog No. 110081) on the label where the color of the reaction zone most closely matches It is determined using the color area. The corresponding result is read in mg / l H ₂ O ₂ units or an intermediate value is estimated if necessary.

Dissolve in methanol at 50/50 w / w to ensure that endogenous hydrogen peroxide is not available in the honey. Measurement in organic solvent (easily volatile ether): The reaction zone of the test strip is immersed in the measurement sample (15-30 ° C.) for 1 second. After evaporation of the solvent (3-30 seconds, gently rub the test strip back and forth), immerse the reaction zone in distilled water for 1 second and allow excess liquid to pass from the long end of the test strip onto the absorbent paper towel. Or gently breathe 4 times over the reaction zone, 3-5 seconds each time. After 15 seconds (catalog number 110011) or 5 seconds (catalog number 110081), the color of the reaction zone is determined using the color area on the label that most closely matches. The corresponding result is read in mg / l H ₂ O ₂ units or an intermediate value is estimated if necessary.

Results FIG. 1a shows the results of the well diffusion assay. In this assay, pasteurized Ulmo honey, pasteurized Ulmo honey (0.1%, 0.01%, 0.01%, 0.0015 or 0.0001%) with different amounts of glucose oxidase enzyme preparation added. w / w), or samples containing Manuka UMF25 + honey were added to wells of agar plates seeded with Staphylococcus aureus. This result shows the diameter of the blocking circle recorded for each sample.

  The antimicrobial effect on Staphylococcus aureus of activated honey containing 0.001% w / w or more glucose oxidase (240 iu / mg) was comparable to that of Manuka 25 + honey.

  The effect of activated honey is bactericidal.

  FIG. 1b shows the results of the well diffusion assay. In this assay, samples containing pasteurized Ulmo honey supplemented with 0.5% w / w glucose oxidase enzyme preparation were prepared as MGO Manuka honey, Manuka 25+ honey, and a series of different phenol standards (10% to 40%) samples. This result shows the area of the inhibition circle of each sample.

  The above results show that the effect of activated Ulmo honey is equivalent to 30% phenol standard, more than twice the effect of MGO Manuka honey and almost twice the effect of Manuka UMF25 + honey. Yes.

  FIG. 2 shows that the potency of activated honey can be adjusted to meet the requirements by adding a suitable amount of enzyme.

Example 3
This example describes the results of a test that demonstrates the antibacterial effect of activated Tineo honey. This honey is described as “activated” if it contains added glucose oxidase.

Well diffusion assays were performed as described in Example 2 using samples of TINEO honey and various phenol standards. FIG. 3a is a photograph of a duplicate agar plate showing the staphylococcus aureus inhibition circle produced by the various samples. This figure contains a drawing showing the placement of the sample in the well of the agar plate.
1.10% phenol standard 2.20% phenol standard 3.30% phenol standard Tineo honey (pasteurized Tineo honey without added glucose oxidase)
5). Tineo inactivation (pasteurized Tineo honey with further heat inactivation)
6). Manuka UMF25 +
7). Tineo 20+ (pasteurized Tineo honey with added glucose oxidase (0.005% Biozyme enzyme))

  Figure 3a shows that neither the Tineo honey alone or the Tineo inactivated honey resulted in inhibition of Staphylococcus aureus growth. The effect of Tineo honey with added glucose oxidase (Tineo 20+) was greater than that of Manuka UMF25 + and was equivalent to the 30% phenol standard.

FIG. 3b is a photograph of an agar plate showing the staphylococcus aureus inhibition circles produced by the various samples. The various samples were arranged as shown below.
1.10% phenol standard 2.20% phenol standard 3.30% phenol standard Tineo honey activity 25+ (0.005% enzyme w / w)
5). Tineo honey activity 25+ (0.005% enzyme w / w)
6). Tineo honey activity 40+ (0.05% enzyme w / w)
7). Manuka Honey UMF25 +

  FIG. 3b shows that samples with Tineo honey activity 25+ and 40+ were more effective in inhibiting Staphylococcus aureus growth than samples with Manuka honey 25+.

Example 4
This example describes the results of tests that demonstrate the antibacterial effect of activated Ulmo and Tineo honey.

  FIG. 4a shows the results of three replicates of the well diffusion assay in which the inhibition circle of Staphylococcus aureus growth was measured after 24 hours of incubation. This assay was performed as described in Example 2. Samples of phenol in deionized water with a concentration range of 40-10%, samples of two manukas (Manuka UMF25 +, Manuka 550 MGO), and samples of activated Ulmo honey with 0.01% w / w Biozyme glucose oxidase (Same as Example 2).

  FIG. 4b shows the results of a further well diffusion assay. Glucose oxidase (Biozyme formulation described in Example 2) was added to pasteurized Ulmo honey (0.005% enzyme w / w) and stored at 40 ° C. for 24 or 72 hours. Stored activated honey samples were then tested for activity against Staphylococcus aureus in a well diffusion assay as described in Example 2 and compared to a series of 4-25% phenol standards. FIG. 4b shows that the activated honey was stable when stored at 40 ° C. for 72 hours, and that the activated honey was comparable to the 20% phenol standard in efficacy against Staphylococcus aureus.

  FIG. 4c shows the results of a further well diffusion assay, in which a new and old batch (old batch) of pasteurized Ulmo honey (Biozyme formulation described in Example 2, 0.5% w / w) is shown. Was received about 4 months before the new batch), and Tineo honey (Biozyme formulation described in Example 2, 0.0005-0.5% w /) increasing the amount of glucose oxidase added. The activity of w) was compared to Manuka 18+ and 25+ UMF honey and a series of 10-40% phenol standards. In FIG. 4c, “normal” refers to as-received pasteurized honey, “inactivation” refers to pasteurized honey undergoing further heat inactivation, and “reactivation” refers to further heat inactivation. This refers to pasteurized honey to which glucose oxidase has been added after conversion. Therefore, Ulmo honey was tested without added glucose oxidase ("Ulmo old 4 normal", "Ulmo new X" and "Ulmo new Y") and with added glucose oxidase. In FIG. 4c, the area of the stop circle of Staphylococcus aureus is shown. The old batch of pasteurized Ulmo honey ("Ulmo Old 4 Reactivation") retains the same degree of activity as the new pasteurized batches ("Ulmo New X Reactivation" and "Ulmo New Y Reactivation") You can see that All of these batches showed about 2-fold higher activity than Manuka honey 18+ and higher activity than Manuka honey 25+. As the amount of glucose oxidase added to pasteurized Tineo honey was increased, the activity of honey increased.

  In this test, 0.005% w / w enzyme is roughly equivalent to 10% phenol standard, 0.05% w / w enzyme is roughly equivalent to 15% phenol standard, 0.5% The w / w enzyme was approximately equivalent to the 25% phenol standard. Manuka 18+ honey was equivalent to a 10% phenol standard, and Manuka 25+ honey showed an intermediate activity between 10% phenol standard and 15% phenol standard.

Example 5
In this example, the effects of different glucose oxidase enzyme formulations were tested using pasteurized Tineo honey. The enzyme formulations used were the Biozyme formulation described in Example 2, and ANHUI MINMETALS DEVELOPMENT I / E CO. , LTD (hereinafter referred to as “Anhui” enzyme preparation), a food grade non-GMO origin glucose oxidase.

FIG. 5a shows the results of a well diffusion assay performed as described in Example 2, in which the enzyme-free pasteurized Tineo honey, or 0.005-0.05% w / w Samples of pasteurized Tineo honey containing two Anhui enzymes were compared to various phenol standards (10%, 20%, 30%) and Manuka honey UMF25 + samples. This result indicates that increasing the amount of enzyme increases the effectiveness of activated honey against Staphylococcus aureus. 0.0125%
The w / w enzyme was approximately equivalent to the 20% phenol standard and the Manuka honey sample, and the 0.05% w / w enzyme was approximately equivalent to the 30% phenol standard.

  FIG. 5b shows the results of a well diffusion assay performed as described in Example 2, in which different amounts of enzyme were added to samples of a commercial batch of pasteurized Tineo honey. The “Tineo” sample is pasteurized honey without added glucose oxidase. “Tineo inactivated” samples are pasteurized honey undergoing further heat treatment and do not contain added glucose oxidase. The “Tineo 20+” sample contains 0.005% w / w Biozyme glucose oxidase. This result indicates that the Tineo sample has no activity against Staphylococcus aureus. The Tineo 20+ sample was more effective against Staphylococcus aureus than Manuka 25+ and was between the 20% and 30% phenol standards.

Example 6
This example describes the stability test results of activated honey stored for 60 days or 90 days.

  To pasteurized Ulmo or Tineo honey, glucose oxidase enzyme from Biozyme described in Example 2 was added at 0.005% w / w and the resulting mixture was added at 37 ° C. for 2 months (Ulmo honey sample) or at room temperature. Stored for 90 days (Tineo honey sample). The amount of enzyme used corresponds to the amount used in commercial food products.

  After storage, the samples were tested in a well diffusion assay against Staphylococcus aureus performed as described in Example 2. Stored samples were compared to 10%, 20% and 30% phenol standards. FIG. 6a shows the results for the Ulmo honey sample, and FIG. 6b shows the results for the Tineo honey sample. Neither type of honey had any apparent loss of activity during storage.

Example 7
In this example, the test results of the antibacterial activity of powder activated honey in which a glucose oxidase enzyme in powder form is added to powdered honey will be described.

  Powdered honey was obtained from Honi Bake from ADM Specialty Ingredients. 0.1% w / w Biozyme glucose oxidase (as described in Example 2) was added to the powdered honey. This mixture was tested for antimicrobial activity in a well diffusion assay against Staphylococcus aureus performed as described in Example 2. The mixture was added to two different wells on the agar plate. The control was used with only honey powder. The result is shown in FIG. The control showed no activity, but the activated honey showed a clear circle of inhibition.

  FIG. 7b shows the results of another well diffusion assay, in which various powdered honeys were obtained from ANHUI MINMETALS DEVELOPMENT I / E CO. , Mixed with powdered enzyme from LTD (Example 5). Sample well 24a is honey powder only, and sample well 24b is honey powder mixed with 1% w / w enzyme. Again, you can see an obvious stop circle.

Example 8
This example shows the antibacterial activity of various different products alone and the test results of the antibacterial activity when they are mixed with activated honey (ie, honey with added glucose oxidase). Antibacterial activity against Staphylococcus aureus was tested in a well diffusion assay performed as described in Example 2. The various samples tested are listed in Table 2 below, and a photograph of the results is shown in FIG.

  This result clearly shows that the antibacterial effect is increased when the product is mixed with activated honey.

Example 9
This example shows the antibacterial activity of various different products alone and further test results of the antibacterial activity when they are mixed with activated honey (ie, honey with added glucose oxidase). Antibacterial activity against Staphylococcus aureus was tested in a well diffusion assay performed as described in Example 2. The various samples tested are listed below and a photograph of the results is shown in FIG.

Figure 9a:
1. Famous brand disinfectant cream number 1
2. Famous brand disinfectant cream number 1 + 10% Ulmo 40+
3. Famous brand disinfectant cream number 2
4). Famous brand disinfecting cream number 2 + 10% Ulmo 40+
5). Tineo honey activity 20+
6). Manuka Honey 25+
7). Ulmo honey activity 20+

FIG. 9b:
1. 1. Cough syrup Cough syrup + 10% Ulmo 40+
3. Famous brand hand cream 4. Famous brand hand cream + 10% Ulmo 40+
5). Tineo honey activity 20+
6). Manuka Honey 25+
7). Ulmo honey activity 20+

  This result clearly shows that the antibacterial effect is increased when the product is mixed with activated honey.

Example 10
In this example, a method for sterilizing a composition of the present invention comprising honey that has not been pasteurized and added purified glucose oxidase is described.

  Ten sealed sachets each containing 50 g of the above composition were gamma irradiated with a target dose of 11.6 to 14.2 kGy (when measured with a dosimeter, the dose was 13.1 to 13.6 kGy), Subsequently, sterility was tested separately.

  In sterility testing, all work was performed under laminar flow in a clean room. 10 g of the above sample was added to 100 ml of sterile tryptone soy broth (TSB: casein pancreatic juice digest, 17 g / L, soybean meal papain digest, 3 g / L, sodium chloride, 5 g / L, dipotassium hydrogen phosphate, 2. 5 g / L, glucose 2.5 g / L, pH 7.3 ± 0.2), shaken and mixed, then transferred to a sterile container. An additional 100 ml of TSB was added to remove all sample residue and added to the same container. TSB was added to the sample and incubated at 30 ° C. ± 2 ° C. for a minimum of 14 days and visually observed for signs of microbial growth. Positive controls were performed on all media before the start of the test.

  At the end of the incubation period, one positive result was noted. Demonstration of a sterilization dose of 35 kGy was observed.

  Testing sachets before and after sterilization by gamma irradiation using a well diffusion assay similar to the assay described in Example 2 confirmed that the effect of irradiation on the antimicrobial activity of the composition was negligible. There was no observable decrease in activity level after sterilization.

Example 11
This example illustrates the results of a test demonstrating the antibacterial effect of the composition of the present invention as compared to Manuka UMF25 + honey and heat inactivated honey (inactive honey). This composition contains honey that has not been pasteurized and added purified glucose oxidase (referred to as "Surgihoney") and is sterilized using gamma irradiation.

  Well diffusion assays as described in Example 2 were performed using Manuka Honey UMF25 +, inert honey and Surgihoney samples. FIG. 10 (a) is a photograph of an agar plate showing the inhibition circle of Staphylococcus aureus (ATCC 9518) produced by various samples. In the figure, Manuka honey UMF25 + is at the top of the plate, inactive honey is at the middle, and Surgihoney is at the bottom. FIG. 10 (b) shows a plate treated with Manuka UMF25 + honey (top) and a plate treated with Surgihoney (bottom).

  This result clearly shows that Surgihoney retained significant antibacterial activity against Staphylococcus aureus after sterilization, and that Surgihoney was more effective against Staphylococcus aureus than Manuka honey UMF25 +.

Example 12
A stability test of a composition of the invention comprising honey that has not been pasteurized and added purified glucose oxidase and sterilized using gamma irradiation (at a minimum dose of 35 kGy).

  Accelerated aging technology is based on the assumption that the chemical reactions involved in material degradation follow the Arrhenius reaction rate function. This function states that when the temperature is increased or decreased by 10 ° C. in a uniform process, the chemical reaction rate changes by about 2 or 1/2 times. For example, 5.3 weeks at 55 ° C. corresponds to one year of shelf storage, 10.6 weeks at 55 ° C. corresponds to two years, and 26.5 weeks corresponds to five years.

  Products from two different production batches were used in this study. The product is a sachet containing 10 g of the above composition. The pouch is sterilized by gamma irradiation with a minimum dose of 35 kGy. Samples were stored at accelerated aging conditions of 55 ° C. (± 2 ° C.).

The relationship between real time and accelerated aging is as follows.

Tests, test interval and required samples Samples from each batch were tested at the same time interval. The following table summarizes the total number of samples required at each time point for the tests performed and for each batch.

Test method Filled sachet weight: The average tare weight of an empty sachet is 1.7 g. Ten sachets were individually weighed on a calibrated laboratory balance and the results were recorded by subtracting the tare weight.

  Pressure test: Each sachet was tested with a pressure tester according to standard operating procedures. The number of passes and failures was recorded.

  Honey pH: 5 bags of contents were placed in a glass beaker. Three standards were used to calibrate the Hanna pH meter, and then the electrodes were rinsed in deionized water. The pH electrode and electrode probe were immersed in a honey sample, and the pH value and temperature were read from a digital display and recorded.

  Water level: Five bags of honey were taken at each time point. Approximately 1 ml of sample was taken from each pouch and placed on the refractometer sample plate (one sample at a time). The user observed through the lens while closing the sample cover and pointing the device at a light source such as a window. The scale values indicated by the shadow lines were read and the results recorded.

  Color: 1 sachet of honey was opened and a small amount of honey composition was placed on a white tile. The color of the honey composition was compared to the Honey Color Chart Pfund Scale and a score was recorded (30-800).

Result Filled sachet weight-accelerated aging

Pressure test results-accelerated aging

pH test results-accelerated aging

Water content test results-accelerated aging

Color test results-accelerated aging

  In Examples 13-35 below, wounds using a composition of the invention comprising honey that has not been pasteurized with added glucose oxidase (in these examples, this composition is referred to as “Surgihoney”) are used. Explains treatment results. Surgihoney provided a sealed sachet with each sachet containing 10 g of composition. The pouch was sterilized using gamma irradiation. The pouch was used from the 0th day of treatment. Every time the dressing was changed, a new Surgihoney was used. The dressing was changed every day as recorded in the examples, or more frequently. Surgihoney was used as a dressing or applied directly to a wound and then covered with a dressing. In both cases, Surgihoney was in direct contact with the wound and was covered with a dressing.

Example 13
In this example, treatment results for infectious footpads using Surgihoney are described. The results are shown in FIG.

The patient was a 78 year old diabetic man. The left foot wound occurred over a month prior to the start of treatment and caused mild pain.
a) Day 0-Wound profile: Wound: 3 x 2 x 1 cm; 99% healthy granulation but 1% green colonization; peeling of surrounding skin; small amount of yellow pus exudate producing odor;
b) Day 5-Wound profile: Wound improvement; Each time honey pouch is used, second dressing change; Metronidazole and Amoxicillin; Tea tree oil is also used;
c) Day 10-Wound profile: wound improvement; disappearance of green colony formation; wound was washed, dry skin removed and additional honey sachets were used.

Example 14
This example describes the treatment results for footpad ulcers using Surgihoney. The results are shown in FIG.

The patient was a 61-year-old diabetic woman and had an infectious footpad ulcer in her right leg that had occurred for a month before treatment and caused mild pain.
a) Day 0-Wound profile: Wound: 2 x 2 x 0.2 cm; 1% tan scab, rest granulation tissue; low amount of yellow exudate;
b) Day 7-Wound profile: wound improvement; using Surgihoney 0.5 pouch with daily dressing change; Flucox;
c) Day 10-Wound profile: Wound further improvement; Flucox discontinued.

Example 15
This example describes the treatment results for footpad ulcers using Surgihoney. The results are shown in FIG.

The patient was a 61 year old diabetic woman who had a foot ulcer on both the first and second footpad of the left foot. Mild pain.
a) Day 0-Wound profile: Wound: 0.8 x 0.8 x 0.2 cm; 100% granulation tissue; Small amount of serous exudation;
b) Day 7-Wound profile: wounds healed to an extent not already open; honey sachet half each day; Flucox;
c) Day 10-wound profile: wound improvement; now dry, no scab or exudate; flucox discontinued.

Example 16
In this example, treatment results for foot ulcers using Surgihoney are described. The results are shown in FIG.

The patient was a 50 year old diabetic woman who had a foot ulcer that occurred in the month prior to treatment. The skin was fragile but no pain was reported.
a) Day 0-Wound profile: Wound: 1 x 0.5 x 0.5 cm; 1% tan scab, rest granulation tissue; very low amount of yellow exudate;
b) Day 7-Wound profile: wound improvement; dry; scab was replaced by healthy granulation; size and depth decreased; use honey dressing (0.5 sachet) after wound wound with saline No antibiotics;
c) Day 9-Wound profile: greatly improved wound; almost closed, no exudate present; subsequent use of dressings with 0.5 sachets each.

Example 17
In this example, treatment results for foot ulcers using Surgihoney are described. The results are shown in FIG.

The patient was 87 years old, a diabetic male with peripheral vascular disease, and had a diabetic foot ulcer that had developed for over a year. The surrounding skin was fragile, but no pain appeared.
a) Day 0-wound profile: wound: 1 x 1.8 cm; 1% tan scab, rest granulation tissue; small amount of yellow serous effusion;
b) Day 3-Wound profile: Wound improvement; Honey sachet used with dressing change; No pain reported; No antibiotics;
c) Day 6-Wound profile: wound improvement; wound bed is now 100% granulation tissue; additional honey sachets used; Diprosalic on dry skin around ulcers.

Example 18
This example describes the treatment results for diabetic foot ulcers using Surgihoney. The results are shown in FIG.

The patient was male and had a diabetic foot ulcer caused by the stimulating action of low quality shoes. The patient had an ulcer less than one month prior to treatment.
a) Day 0-wound profile: red surrounding skin and mild pain; 1% scab, 99% granulation; infection and diabetes; low amount of yellow serous effusion; wound size: 2 cm x 2 cm x 0. 2 cm;
b) Day 7-Wound profile: Diagnosis in the hospital is over, but dressing is changed daily from day 1 to the present day. The dressing material was changed by the son's wife, a regular nurse, according to the procedure manual. Wound improvement; 100% healthy granulation; healthy surrounding skin, mild pain; antibiotics used, Flucocacillin, 500 mg, every 6 hours for 7 days; wound size: 1.5 cm × 1.3 cm × 0.1.

Example 19
In this example, treatment results for foot ulcers using Surgihoney are described. The results are shown in FIG.

The patient was a 55-year-old male with severe and poorly managed diabetes. This patient had a deep ulcer at the site of the third footpad after amputation.
a) Day 0-Wound profile: Wound: 3 x 3 x 3 cm; 1% tan scab; 1% green colony formation; rest granulation tissue; medium amount of yellow pus exudate;
b) Day 1-Wound profile: wound improvement; all scabs removed before use of honey (1 sachet); metronidazole;
c) Day 3-Wound profile: wound improvement; use of additional sachets; patient discharge; wipe specimen: +++ skin microflora; no antibiotics.

Example 20
This example describes the treatment results for traumatic leg wounds using Surgihoney. The results are shown in FIG.

The patient was a 95-year-old woman who had a traumatic wound in her lower leg that caused moderate pain.
a) Day 0-Wound profile: Wound: 15 x 12 x 1 cm; mostly granulation tissue, but 1% of the wound bed is necrotic and black; medium amount of bloody exudate;
b) Day 3-Wound profile: no wound change; no further necrosis but the wound is about the same; honey 1 sachet used; wiped specimen: Entrecoccus sp. No antibiotics;
c) Day 8-Wound profile: wound improvement; reduced exudate volume and reduced necrotic tissue.

Example 21
This example describes the treatment results for infectious leg wounds using Surgihoney. The results are shown in FIG.

  The patient was 91 years old and had an infectious wound on her lower leg. The surrounding skin was fragile and there was mild pain.

a) Day 0-Wound profile: wound size: 4.5 x 2.5 cm; most red granulation tissue and 1% yellow / brown scab; medium amount of yellow serous effusion.
b) Day 15-wound profile: wound improvement; size: 4 x 2 cm; less scab and serous exudation;
c) Day 19-Wound Profile: Wound Improvement; Use Surgihoney with each dressing.

Example 22
In this example, treatment results for foot ulcers using Surgihoney are described. The results are shown in FIG.

The patient was a 53-year-old diabetic woman with chronic diabetic foot ulcers that persisted after amputation of the first, second, third and fourth toes.
a) Day 0-wound profile: wound: 6.5 x 3 x 0.2 cm; granulation tissue and low amount of yellow serous effusion;
b) Day 5-Wound profile: wound size slightly improved but wound bed condition changed little; unbound granulation tissue removed; honey sachets used with absorbent dressing; antibiotics No substance;
c) Day 7-Wound profile: Wound lavage, with healthy granulation tissue; Change dressings using honey sachets each time every 2 days;
d) Day 9-Wound profile: wound improvement; size decreased, exudate decreased; patient discharged.

Example 23
In this example, treatment results for leg ulcers using Surgihoney are described. The results are shown in FIG.

The patient was a woman who had a leg ulcer that had developed over the year before the start of treatment and had mild pain.
a) Day 0-wound profile: healthy surrounding skin; 1% scab, 99% granulation; moderate amount of yellow serous effusion; wound size: 7 cm x 3 cm;
b) Day 7-Wound profile: Wound improvement, patient used community care; Surgihoney used one sachet; decreased in size and reduced exudate present; became more tolerable pain.

Example 24
In this example, treatment results for leg ulcers using Surgihoney are described. The results are shown in FIG.

  The patient was a 77-year-old male who was malnourished and dehydrated. anemia. Prostate cancer. The patient had a leg ulcer that had developed over 6-12 months prior to treatment. Mild pain.

a) Day 0-Wound profile: Wound: 20 x 10 x 0.5 cm; 1% tawny scab, rest is granulation tissue but has moderate green pus exudate; Wipe specimens: ++ Coliform and Pseudomonas Antibiotics: Ben Pen and Clinda;
b) Day 4-wound profile: no wound change; honey-only dressing; Ben Pen and Clinda;
c) Day 8-Wound profile: wound improvement; all green scabs disappeared; no antibiotics; paracetamol.

Example 25
In this example, treatment results for leg ulcers using Surgihoney are described. The results are shown in FIG.

  The patient was a 91-year-old woman with leg ulcers that had developed over 6-12 months. She was hospitalized and had severe pain.

a) Day 0-wound profile: severely painful and odorous wound; infection; scab 10%, green colonization 50%, cellulitis 20%, healthy granulation 20%; wound: 15cm x 5cm x 0. 3 cm; exudate: low volume, yellow serum;
b) Day 11-Wound profile: mild pain, greatly improved wound; 100% healthy granulation; no infection; wound: 10 cm x 5 cm x 0.2 cm; exudate: low volume, bloody red; wound size 50% decrease.

Example 26
In this example, treatment results for leg ulcers using Surgihoney are described. The results are shown in FIG.

The patient was a 78-year-old male patient with urinary tract infection. This patient had an infectious leg ulcer. The surrounding skin was macerated. The wound produced mild pain.
a) Day 0-wound profile: wound size: 5 cm x 6 cm x 0.5 cm; red granulation 98%, yellow / brown scab 1%, necrotic tissue 1%; low amount of bloody red exudate;
b) Day 8-Wound profile: wound improvement; use 1 sachet of honey for each dressing change; wiped specimen reveals slight skin microflora; no antibiotics.

Example 27
In this example, treatment results for leg ulcers using Surgihoney are described. The results are shown in FIG.

The patient was a 82-year-old woman who was malnourished and dehydrated. The patient had a leg ulcer that had developed over 3 months prior to treatment.
a) Day 0-wound profile: wound: 3 x 3 x 0.5 cm; wound bed consisting of healthy granulation tissue; small amount of yellow blood exudate;
b) Day 3-wound profile: wound improvement; wiping specimen reveals ++ skin microflora; honey 1 sachet with dressing; no antibiotics;
c) Day 8-Wound profile: wound improvement; wound size decreased and exudate volume decreased.

Example 28
In this example, treatment results for leg ulcers using Surgihoney are described. The results are shown in FIG.

The patient was an 83 year old, incapable man with low serum albumin. The patient had leg ulcers that had developed over the three months prior to treatment and had moderate levels of pain.
a) Day 0-wound profile: wound: 20 x 18 x 0.5 cm; yellow / brown scab 1%, green colony formation 1%, the rest granulation tissue; medium amount of green serous exudation; slight wound odor;
b) Day 4-Wound profile: wound improvement; using 2 sachets of Surgihoney; + Pseudomonas by wiping specimen; no antibiotic intake;
c) Day 8-Wound profile: Wound improved after two changes of honey-only dressing; +++ Pseudomonas; markedly improved sufficiently and honey discontinued.

Example 29
In this example, treatment results for leg ulcers using Surgihoney are described. The results are shown in FIG.

  The patient was a 74-year-old man who smoked, suffered from type 2 respiratory failure and had poor exercise. This patient had a septic skin laceration that had occurred over the week prior to treatment and had moderate levels of pain.

a) Day 0-Wound profile: Wound: 15 x 20 x 0.3 cm; 99% healthy granulation but 1% infected at risk; medium yellow exudate; Found; antibiotic: clindamycin;
b) Day 6-Wound profile: Wound improved after two changes of dressing; Honey-only dressing; Ulcer is now dry; No growth in wipes; Clindamycin;
c) Day 9-Wound profile: wound improvement; cellulitis still remains but ulcer healed; no growth in wipe again specimen; clindamycin.

Example 30
In this example, treatment results for leg ulcers using Surgihoney are described. The results are shown in FIG.

The patient was an 81-year-old male with heavy and extensive ulcers on the legs and arms. The patient suffered from peripheral vascular disease and was malnourished. The wound had occurred over the month before treatment. The surrounding skin was peeled off, causing severe pain.
a) Day 0-wound profile: wound: 30 x 28 x 1 cm; yellow / brown scab 1%, the rest granulation tissue; large amount of red blood exudate; wiped specimen reveals a mixture of ++ E. coli and Pseudomonas;
b) Day 6-Wound profile: wound improvement; slight Pseudomona; no antibiotics but for pain oromorp;
c) Day 10-Wound profile: wound improvement; + Pseudomonas still present; severe pain decreased.

Example 31
In this example, treatment results for leg ulcers using Surgihoney are described. The results are shown in FIG.

The patient was 67 years old, a man with peripheral vascular disease. Leg ulcers had developed over a week prior to treatment. The surrounding skin was fragile and the pain was moderate.
a) Day 0-wound profile: wound: 1 x 1 x 0.2 cm; yellow / brown scab 1%, the rest granulation tissue; small amount of yellow serous exudation in the wound;
b) Day 11-Wound profile: now showing signs of improvement after a series of dressings changes where there was no change in wound; exudate decreased and wounds reduced; one honey sachet with each change A wiping specimen reveals staph aureus ++;
c) Day 18-Wound profile: no wound change; wound size: 3 x 3 x 0.2 cm; staph aureus ++ present.

Example 32
In this example, treatment results for leg ulcers using Surgihoney are described. The results are shown in FIG.

  The patient was a 43-year-old woman with a large diabetic ulcer that existed for 1 month. The ulcer was associated with latent osteomyelitis and the bone was wound excised and metallized from the ribs. The patient had a broken leg and subsequent treatment resulted in a bone infection. The leading clinician's opinion was that this patient was likely to have a foot amputation.

a) Day 0-Wound profile: 40% scab cover, 10% green colony formation, 50% healthy granulation wound; healthy surrounding skin; moderate pain; moderate amount of yellow pus exudate; wound 9 cm x 8 cm x 2 cm No antibiotics;
b) Day 1-Wound profile: no wound change; wiped specimen: E. coli and Pseudomonas; no antibiotics; no analgesics;
c) Day 14-Wound profile: wound improvement; using one sachet of Surgihoney; wound slowly granulates and greatly improves; wiped specimen: Slight skin microbiota found.

Example 33
This example describes the results of treating pressure ulcers using Surgihoney. The results are shown in FIG.

The patient was 88 years old and a diabetic woman who was malnourished. This patient had pressure ulcers (grade 3) that had developed for 6 months prior to treatment. Medium level of pain occurred and surrounding skin was fragile.
a) Day 0-wound profile: wound size: 0.7 x 0.7 x 0.2 cm; yellow / brown scab 1%, cellulitis tissue 1%, the rest granulation; low amount of yellow pus exudation liquid;
b) Day 3-Wound profile: wound healing (wound closed); use one sachet of Surgihoney.

Example 34
This example describes the treatment results for infectious axillary wounds using Surgihoney. The results are shown in FIG.

The patient was a 21-year-old woman who had an infectious wound under his heel. The surrounding skin was red and the pain was mild.
a) Day 0-wound profile: cellulitis 1%, granulation 99%; low amount of yellow serous effusion; wound size: 5 cm x 5 cm;
b) Day 24-Wound profile: wound improvement; on holiday, using honey for 2 weeks; no antibiotics.

Example 35
This example describes the results of treatment of infection around the catheter entrance using Surgihoney. The results are shown in FIG.

The patient was a 41-year-old woman with cancer. The patient had an infection of the breast area around the catheter entrance. The wound was present for less than 1 week prior to treatment and had mild pain.
a) Day 0-wound profile: wound: 2 cm x 2 cm; cellulitis 1%, the rest granulation tissue; low amount of red serous effusion;
b) Day 6-Wound profile: Wound improved greatly; no exudate; one sachet of Surgihoney; wiped specimen but no growth; no antibiotics;
c) Day 14-Wound profile: Wound improves to a point where it is no longer a problem.

Example 36
Surgihoney
Surgihoney is an unpasteurized honey with added glucose oxidase. Three different Surgihoney formulations with different antibacterial activity were made.

  S1 Surgihoney: Unpasteurized honey with 0.1% (w / w) added glucose oxidase. The enzyme used was food grade glucose oxidase from Aspergillus niger manufactured by BIO-CAT INC, and the activity was 15,000 units / g. Sealed pouches containing 50 g of S1 Surgihoney were gamma irradiated at a target dose of 11/6 to 14.2 kGy.

  S2 Surgihoney: Unpasteurized honey with 0.1% (w / w) added glucose oxidase. The enzyme used was glucose oxidase (GO3B2) from Aspergillus niger manufactured by BBI Enzymes Limited, and the activity was 274 units / mg. Unit definition: The amount of enzyme that oxidizes 1 micromole of glucose per minute at 25 degrees Celsius, pH 7.0. Contaminants: alpha amylase 0.05% or less, saccharase 0.05% or less, maltase 0.05% or less, and GO / Cat2000 or more.

  S3 Surgihoney: Unpasteurized honey with 0.25% (w / w) added glucose oxidase. The enzyme used was glucose oxidase (GO3B2) manufactured by BBI Enzymes Limited, and the activity was 274 units / mg.

  Thus, S1 Surgihoney has 15 units of glucose oxidase per gram of composition, S2 Surgihoney has 274 units of glucose oxidase per gram of composition, and S3 Surgihoney has 685 units of gram per composition. Contains glucose oxidase.

Example 37
In vitro antibacterial activity of Surgihoney In this example, the sensitivity of a series of wound and ulcer bacterial isolates to Surgihoney by disc diffusion method, determination of minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC), and time bactericidal measurement Explain the test.

Summary Results: Surgihoney shows very potent inhibitory and bactericidal activity against a wide range of gram positive and gram negative bacteria and fungi. The MIC / MBC is significantly lower than the concentration expected to be reached with local clinical use. The local concentration of Surgihoney in the wound is estimated to be about 500 grams / L. Staph. Surgihoney 1 MIC / MBC for Aureus is 31 and 125 grams / L and Surgihoney 3 MIC / MBC is 0.12 and 0.24 grams / L.

  The sterilization rate depends on the efficacy. In the least potent Surgihoney 1, complete bactericidal activity occurred within 48 hours for all microorganisms tested. With the most potent Surgihoney 3, bactericidal activity occurred within 30 minutes. Maintenance of the Surgihoney inoculum formulation within 1 week showed complete bactericidal activity and no bacterial residues.

  Conclusion: Surgihoney has broad potential as a highly active topical treatment that combines the healing properties of honey with the powerful antimicrobial activity of bioengineered products for skin lesions, wounds, ulcers and cavities Yes. High activity against multi-drug resistant bacteria. It is more active than the other honeys tested and is comparable to chemical disinfectants in antibacterial activity.

  Superficial wounds and skin ulcers are becoming more and more common with the aging of the population in many countries and with the global epidemic of obesity and type 2 diabetes. In the UK, community nurses spend a lot of time on leg ulcer care, and if they want to maintain standards in their local leg ulcer services, supervision by leg ulcer nurses is essential . Chronic wounds on the skin are often colonized with bacteria. Although it is difficult to know when they become pathogenic and whether they are pathogenic, bacterial colonization delays tissue healing, even if no obvious infection is present, It is believed to play a role in creating a biofilm and, as a result, creating a wound scab and malodor.

  Tissue survival is particularly difficult when combined with comorbidities. Chronic wounds are always colonized with bacteria, which can destabilize the healing process. If a microbial sample is reported to be a growing bacterium, it will send the sample and attempt to provide whole body antibiotics. This only contributes to selecting even more resistant microorganisms, which is why chronic leg ulcers are so frequently colonized with multi-drug resistant microorganisms such as methicillin-resistant Staphylococcus aureus and Pseudomonas aeruginosa That is why.

  Surgihoney has been developed as a prophylactic dressing for wounds. In this study, we examine the in vitro properties of Surgihoney. Surgihoney retains all of the established natural honey healing properties, but its antimicrobial activity can be set at whatever degree of efficacy is required. In this study, the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of Surgihoney 1, 2 and 3 and the time bactericidal curve were measured.

Methods Surgihoney was offered as efficacy grades 1, 2 and 3. Surgihoney is given in a semi-solid form as a sterile pharmaceutical grade in a sachet.

  Clinical isolates were recovered from soft tissue microbiology samples. 18 isolates of Staphylococcus aureus, 12 strains of methicillin sensitive (MSSA) and 6 strains of methicillin resistant (MRSA), 6 isolates of β-hemolytic streptococci, Lancefield group A (2), B (2), C (1 ), G (1), vancomycin resistant E. coli. Enterococcus spp. containing faecium. Including 5 isolates of Escherichia coli, a broad spectrum β-lactamase producing strain coli 6 strain, Klebsiella spp. 2 strain, Serratia marcescens Amp C producing strain 1 strain, Pseudomonas Aeruginosa4 strain, Acinetobacter Lwoffii1 strain Propionibacterium Acnes1 strain, Bacteroides Fragilis1 strain and Candida Albicans2 strain was tested Candida Glabrata1 strain, the Aspergillus Fumigates1 strain against Surgihoney.

Agar Diffusion 6 mm wells were cut in isosenites agar that had already been seeded with test microorganisms at a concentration that gave semi-confluent growth. The test Surgihoney and other honey from the preliminary study were added to the wells.

  Preliminary studies were conducted to compare Surgihoney efficacy S1, S2, S3 with various honeys from all over the world, European honey, South American honey, New Zealand honey, Yemani honey, Sudane honey, and medical honey Medihoney And antibacterial coatings containing silver (Silver Aquacel) and iodine (Iodoflex). Wells were cut out in plates seeded with Staphylococcus aureus and filled with test honey, or in the case of dressings, they were cut 2 × 2 cm and placed on the surface of the seeded plate.

  After preliminary studies, Surgihoney efficacy S1, S2, S3 was tested alone against a series of bacterial isolates from skin lesions. Wells were filled to the surface using a formulation that was emulsified by diluting about 2 grams of three undiluted Surgihoneys with an equal volume of sterile water. After aerobic incubation for 18-24 hours (longer for Candida and Aspergillus spp. And anaerobic for Propionibacterium sp. And Bacteroides sp.), Zone sizes were measured.

Minimum Growth Inhibitory Concentration and Minimum Bactericidal Concentration Surgihoney products were liquefied by warming to 37 ° C., and 5 grams thereof was mixed with 10 mL of sterile deionized water. This dilution was considered as “no dilution” material for serial dilution. The British Antibacterial Chemotherapy Association (BSAC) method for the implementation of minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) was used (Andrews JM. Determination of minimum concentrations. J Antimicrob 372 Chem1 482 Chem; Supp 1): 5-16). Surgihoney products were serially diluted from no dilution to 1024 times in wells of a microtiter tray. 75 μL of each honey dilution was added to each well in the microtiter tray strip. The undiluted concentration showed a concentration of 250 grams / L and the 2048-fold dilution showed about 0.12 grams / L.

  Test microorganisms were prepared by taking 4 colonies from a pure culture that were morphologically identical for each microorganism to produce a 0.5 McFarland density. This was further diluted 1:10.

  All wells containing controls were seeded with 75 μL of test isolate formulation. The well tray was incubated at 37 ° C. for 18 hours. The MIC was considered as the most diluted well showing no detectable turbidity.

  The MIC well and wells near the MIC well were subcultured on a blood agar medium and incubated at 37 ° C. for 18 hours to measure MBC. MBC was the most diluted concentration that showed no growth after incubation.

Time-sterilization curve The test bioinoculum was prepared by taking 0.1 mL of 0.5 McFarland density test organism and incubating it in 3 mL of nutrient broth. The test inoculum was divided into 3 separate Bijou vials, and 0.5 g of Surgihoney 1 (S1), Surgihoney 3 (S3), or Medihoney (MH) was added to the control and 3 test formulations. Inoculum colonies were determined by serial dilutions of 1:10 and plating by 0.1 mL on blood agar plates and repeated three times.

  The test and control inocula were maintained at 30 ° C. to simulate the temperature of the superficial skin lesion. Colony counts were performed as described above in triplicate at 0.5, 2, 4, 24, 48, 72 and 168 hours.

  In terminal culture, inoculate 0.1 ml of the initial inoculum into nutrient broth to neutralize any residual effects of Surgihoney, incubate at 37 ° C for 72 hours, and then plate into blood agar to determine test organism survival. Went by.

Results Inhibition Circle Size Preliminary comparative studies demonstrated that all Surgihoney efficacy had higher antimicrobial activity than any other honey tested, including the medical grade honey honey. The blocking circle for S1 was larger than the blocking circle formed by any other honey. The silver coating material produced some blocking effect under the coating material, but there was no blocking circle as was present in Surgihoney. The iodine coating formed a large blocking circle (about 70 mm) against Staphylococcus aureus, which was larger than S1 (36 mm) and equivalent to S3 (67 mm).

  In a quantitative inhibition circle size test, all-potency Surgihony showed a circle of inhibition in agar diffusion for all bacteria tested, both gram positive and gram negative bacteria, including multiple antibiotic-resistant bacteria, and fungal species. Formed. The inhibition circle size for each increased with increasing Surgihoney composition efficacy. Table 12. Surgihoney's blocking effect is not only dependent on direct contact with the active agent, as with silver coatings, but also diffuses well beyond the well, creating a large blocking circle as listed in Table 12. Formed.

MIC and MBC
Surgihoney showed significant antibacterial activity against all isolates tested. MIC and MBC agreed well among isolates of the same species, regardless of whether the isolate was multidrug resistant or highly sensitive. Table 13 lists the MIC and MBC values for the isolates tested by dilution ratio, and Table 14 shows the MIC and MBC in grams per liter. The degree of efficacy increased with the Surgihoney grade. The MBC for each isolate was close to the MIC and in most cases was within a single dilution.

  The local concentration of Surgihoney in the wound is estimated to be about 500 grams / L. Staph. The MIC / MBC for Surgihoney 1 for Aureus is 31 and 125 grams / L, respectively, and the MIC / MBC for Surgihoney 3 is 0.12 and 0.24 grams / L.

Time sterilization curve Surgihoney quickly kills bacteria. Starting with about 105 colony forming units per milliliter (cfu / mL), the number of cfu / mL in the control continually increased while in the Surgihoney inoculum, after contact with both potencies of Surgihoney, cfu / mL fell rapidly. By 30 minutes, in both S1 and S3, cfu numbers often decreased 1000-fold (FIG. 34). In S1, in many cases, bacterial growth was undetectable by 2 hours, and in S3, it was undetectable by 30 minutes. Enterococci appeared to be stronger and survived for 48 hours. Bactericidal activity was complete for all organisms since terminal cultures in nutrient broth followed by plating on blood agar did not detect any organisms in the S1 or S3 inoculum.

DISCUSSION Surgihoney is a natural honey and, unlike many commercial honeys for human consumption, is also an organic matter in the current sense of having no agricultural additives or antimicrobial residues. Surgihoney does not depend on a specific nectar source, unlike honey, such as manuka, which depends on a specific plant nectar source for high activity. Surgihoney can adjust antibacterial activity through the formulation process, allowing the production of various grades with measured uniform potency.

In this study, the efficacy of Surgihoney was clearly demonstrated to be active against all species of bacteria and fungi tested as very potent antimicrobial agents. In a preliminary study comparing Surgihoney to various honeys from all over the world, and compared to the medical grade honey Medihoney, Surgihoney showed significantly higher antimicrobial efficacy. Compared to the commonly used topical disinfectants silver and iodine, Surgihoney 3 produces an antibacterial effect as high as the iodine coating, and the silver coating (Aquacel)
An antibacterial effect higher than that of Ag) was generated. The silver coating was only effective at inhibiting bacteria that were in direct contact with the coating.

  In MIC and MBC studies, Surgihoney not only blocks microorganisms, but also kills microorganisms at concentrations 10 to 1000 times lower than the concentration expected to be achieved in local therapy (estimated at 500 grams / L). It is shown. The bactericidal activity of Surgihoney occurs at a concentration close to its inhibitory activity. Thus, Surgihoney can be highly active in multimicrobial inhibition and eradication when used topically in any colony wound or superficial infected wound or soft tissue cavity. Many chronic wounds are colonized with resistant bacteria and wound healing is delayed by bacterial residues in biofilm formation. Thus, the use of Surgihony not only promotes wound healing, but may help reduce the use of appropriate antibiotics. In clinical use, the local concentration of Surgihoney at the wound site will be significantly higher than the concentration of systemic antibiotics in serum or deep tissue. This is reflected in Surgihonney's MIC and MBC values, which are correspondingly higher than those generally shown for systemic antibiotics.

  The bactericidal activity rate is shown to be very fast by the time bactericidal curve, which is within 30 minutes for Surgihoney 3 and within 2 hours for Surgihoney 1. This is the case for both gram positive and gram negative organisms, but Enterococci appears to be slightly more robust. Fungi, Candida spp. , Aspergillus sp. However, higher concentrations and longer exposures are required to inhibit growth and kill organisms.

  Surgihoney is used as a topical wound dressing for skin lesions and cavities with the aim of providing a moist wound healing environment while also reducing microbial colonization and assisting in removing scabs and promoting granulation and epithelialization It is formulated as a sterile product.

  Other antibacterial formulations are available as topical formulations intended for the treatment or prevention of wound infections. The silver impregnated coating appears to have good antibacterial activity but also exhibits cytotoxicity when compared to the honey formulation. Iodine analogues also have good antibacterial activity, but have also been reported to be toxic in certain situations. Concerns about using chlorhexidine formulations in wound dressings are also increasing due to the development of antimicrobial resistance and toxicity.

  Surgihoney's clinical utility appears to be topical use on the skin, in wounds and cavities. Wounds can be colonized with bacteria that can form a biofilm and delay healing. Due to increasing concerns about antimicrobial resistance and the shortage of new antimicrobial agents, topical agents with broad antimicrobial activity may play a role in reducing the use of systemic antibiotics in soft tissue lesions. These in vitro studies have demonstrated Surgihoney's ability as a wound dressing with high antimicrobial activity. The efficacy of Surgihoney can be regulated, and Surgihoney also exerts other important functions in wound healing: wet barrier, scab cover, local nutrient supply, local immune regulation and cell toxicity is not.

Conclusion These in vitro results support the clinical use of Surgihoney as a wound dressing and it is not only a powerful and non-toxic antibacterial agent, but also plays all the roles required in the wound healing process. It may be the first product that can be demonstrated.

Example 38
Surgihoney's Antiviral Activity S1 or S2 Surgihoney was mixed with herpes simplex virus in cell culture (50% honey and virus mixture in cell culture) and then incubated at 37 ° C. for 1 hour. This mixture was then plated on cells and the number of virus plaques formed with each mixture was recorded. Controls without honey or with control honey were also performed.

  The number of virus (herpes simplex virus) plaques recorded after 1 hour incubation is shown in FIG. No plaques were formed with the S1 or S2 Surgihoney mixture, compared to 160 plaques with the honey-free mixture and 150 plaques with the control honey mixture.

  This result indicates that both the S1 and S2 Surgihoney formulations have potent antiviral activity.

Example 39
Use of Surgihoney in line site dressings To assess the effectiveness of Surgihoney in preventing infection of peripherally inserted central venous catheters (PICC lines), S1 Surgihoney is applied locally to the line entrance site in the arm of 30 patients. used. Approximately 3-8 g of S1 Surgihoney was used for the dressing, which was then contacted with the wound and kept in place with the second dressing. Line site colonization and line-related bacteremia were assessed and compared to 30 patients who did not receive Surgihony dressing. The results are shown in Table 15 below.

  It was concluded that Surgihoney is an effective antibacterial agent for use in line site dressings.

Example 40
Use of Surgihoney to prevent infection of cesarean section wounds Surgical wound infection is a particular problem in cesarean section, which has a fairly high infection rate of about 10%. In the UK, with 147,726 cesarean sections each year, cesarean wound infections (8-24.6%) and widespread variants across NHS hospitals (13.6-31.9%) (Bragg et al., 2010. Variation in rates of caesarea section among English NHS trusts after accounting for 10 and 20 minutes). Cesarean wound infection is a major cause of long hospitalization, resource consumption, and other morbidity and mortality. Recovery from cesarean section is more difficult in women who develop postoperative wound infections.

  To evaluate the effectiveness of Surgihoney in preventing infection of cesarean section wounds, S1 Surgihoney was used topically once on the wound after surgery. About 25 g to 35 g of S1 Surgihoney was used for the dressing, which was then contacted with the wound and kept in place with the second dressing. Over three months, nearly 200 patients were evaluated.

Clinical Evaluation Women undergoing cesarean section (CS) between October 2012 and January 2013 were offered Surgihoney as a single use wound dressing in covering the wound at the end of the surgical procedure. Each patient used 10 g Surgihoney sachets. Using aseptic technique, a “non-sterile” surgical assistant opened the Surgihoney pouch and carefully applied the sterile contents onto the sterile dressing. The obstetrician or operating room midwife then applied the dressing to the surgical wound. After the surgical procedure, the midwife in charge completed the evaluation record. Data collected were MRSA status, diabetes history, medication and body volume index. For 14 days after the surgical procedure, the midwife in charge also recorded the presence of any wound healing problems, especially exudation, pain and inflammation. If there was inflammation, wound culture wipes were requested and microbiological results were recorded. Surgical site infection (SSI) rates during the 3-month evaluation using Surgihoney coating were compared to infection rates in the 9 months prior to the evaluation based on data collected by the infection control team. Wound infection was clinically defined as an inflammatory wound (erythema, swelling, secretion) requiring antibiotic treatment. The SSI rate was calculated as a percentage of all CS surgical procedures performed.

Results The results are shown in Table 16 below. There were 186 CSs in the three-month period from October 2012 to January 2013, of which 102 (55%) were urgent. None of the women were colonized with MRSA. Four (2.23%) suffered from diabetes. Forty-two (27.3%) had a body mass index greater than 25. During the evaluation, 4 out of 186 cases were confirmed. This indicated an infection rate of 2.15%. One patient reported an adverse event associated with Surgihoney use, which was a form of wound stimulation and resolved in 3 days without further therapeutic intervention. In the last nine months, there were 590 CS surgical procedures (234 were selected, 356 were urgent), and 32 CS SSIs were recorded by infection control observers, which was 5.42% Showed the infection rate. Decrease in infection rate is significant: p = 0.042 (χ ² test).

  The above results indicate that the surgical site infection rate was reduced (60% reduction) in the group treated with S1 Surgihoney compared to historical data. Surgihony dressing was well tolerated and few adverse events were reported.

  When using Surgihoney, the wound infection rate was reduced by 60.33%. Using SSI data from the two groups in this study, the CS SSI rate (expected) before Surgihoney was 5.42% and later 2.15% (observed). At these levels (lower than the previously reported 9.6% infection rate), the estimated CSS SSI infection rate in the UK is 8007 cases per year (expected) and 3176 cases per year (observation) ). The difference that can potentially be reduced by using Surgihoney is 4831 cases.

  It was concluded that S1 Surgihoney effectively reduces the infection rate of cesarean wounds after surgery. The prevention of wound colonization using Surgihoney, a drug that is not toxic to the healing tissue and also accelerates the healing process, is a new and potentially important discovery, which is a surgical discovery. There is a possibility to change the way to deal with. Surgihoney offers a clinically effective and cost-effective therapeutic intervention that significantly reduces SSI in women undergoing cesarean section.

Discussion This evaluation demonstrated that Surgihoney, a highly effective antimicrobial wound dressing, could be used as a wound dressing for early CS wounds to prevent infection. As a “natural” product with established wound healing properties, Surgihoney appears to promote wound healing in addition to providing strong antimicrobial activity to prevent wound colonization and infection. Some halogenated chemical disinfectants may provide similar antibacterial activity but may delay wound healing (Jan WA. Comparison of conventional pydine dressing with honey)
dressing for the treatment of diabetic foot ulcers. JPMI-Journal of Postgraduate Medical Institute 2012; 26 (4): 402-7)). Iodine wound dressings are contraindicated in CS (Joint Formulary
Committee. The British National Formula. London: The Pharmaceutical Press; 2013) and its use is associated with various toxicities (Pietsch and Meakins: Complications of povidone-iodine absorption
in topically treated burn patents. The
Lancet 1976; 307 (7954): 280-2; Scoggin et al .: Hypernatramia and acidosis in associat.
ion with topical treatment of burns. The
Lancet 1977; 309 (8018): 959; Donovan et al .: Seizures in a Patient Treated with Continuous Povidone-Iodine Medianary Irritation. New England Journal of Medicine 1992; 326 (26): 1784; Colpaert: Iodine toxicity as a cause of total triblock block in burnt. Burns 2009; 35: S45-S6; Ramaswamykanive: Cardiovascular collapsing following povidone-iodine wash. Anaesthesia and Intensive Care 2011; 39 (1): 127-30; Lakhal: Povidone iodine: Features of critical system absorption. Analyzes Francais d'Anesshesie et de Reanimation 2011; 30 (7-8): e1-8): e1-e3. ).

  Similarly, in Cochrane systematic reviews, whether silver-containing dressings or topical agents promote wound healing and prevent wound infection (Storm-Verslott et al .: Topical silver for preventing wound anti-wound infection. Cochrane system. Or whether they are an effective treatment for infectious or contaminated chronic wounds (Vermeulen et al .: Topical silver for treating infected wounds (Review). Cochrane reviews 2010; (10): 42) There is no evidence.

  Although previous systematic reviews have shown evidence of ambiguous benefits for the clinical efficacy of honey as a wound dressing, this new formulation would provide significant clinical benefit in CS patients (Jull et al .: Honey as a topical treatment for wounds: The Cochrane Collation, 2009; Jul et al: Honey as a topical treatment for wounds. In a temporal comparison of wound infection rates, the above assessment shows that infection rates have decreased by 60.33%, from 5.42% before treatment intervention to 2.15% when using Surgihoney. Yes.

  Health-related infections are a serious and costly medical complication affecting about 8% of hospitalized patients, SSI accounts for 14% of these infections, and nearly 5% of patients undergoing surgical procedures develop SSI It has been found. SSI is associated with a fairly high morbidity, with more than a third post-operative death associated at least in part with SSI. Antimicrobial prophylaxis is routinely used in many surgical procedures to reduce surgical wound infection. Skin disinfection is also routinely used by surgeons prior to skin incision to reduce skin bacterial load, but the use of antimicrobial dressings is not routinely practiced. The reason for this may be because most topical disinfectants have an adverse effect on tissue healing.

  Surgihoney is a product with strong antibacterial activity that is non-toxic and promotes tissue healing. By using this product locally on “clean” surgical wounds, systemic antibiotic prophylaxis can actually be replaced in certain types of surgery. Such progress will help reduce the amount of antibiotic used and the pressure of colonizing bacteria.

  The cesarean wound was chosen for this assessment because the cesarean section patients were generally healthy, had no or little morbidity, and increased CS infection rates. This is because it has been reported. Possible reasons for this increase are an increase in older mothers, an increase in mothers with comorbidities, especially diabetes, and an overall increase in mothers with higher body mass index. The use of antibacterial agents in early wound dressings has not been routine before, but this evaluation demonstrates the interesting and effective role of Surgihoney in preventing CS wound infection.

Example 41
Use of S1 Surgihoney to Treat Infectious Leg Ulcers The patient was 52 years old, a man with diabetes and chronic kidney disease. The ulcers were widespread, painful, smelly, and developed a large amount of severe exudation.

  Along with antibiotics, S1 Surgihoney was used as a topical dressing. By day 7, the wound odor had diminished, the pain was gone, and there was no massive exudation and pseudomonas infection. The patient was discharged on day 7.

  The resulting photograph is shown in FIG. In FIG. 36, (a) shows the first day of treatment; (b) shows the fourth day of treatment; and (c) shows the seventh day of treatment.

Example 42
Use of S1 Surgihony to Treat Pressure Sores The patient was 50 years old, a female patient suffering from spina bifida, physically disabled and unable to move. This patient had a pressure ulcer from the waist to the sacrum that persisted for over a year. The cavity was infected with Streptococcus pyogenes.

  S1 Surgihoney was used as a topical dressing. From the second day, wound improvement was reported. By day 30, the soft tissue cavity had healed almost completely. At this time, Streptococcus was not detected.

  The resulting photograph is shown in FIG. In FIG. 37, (a) shows the first day of treatment and (b) shows the 30th day of treatment.

Example 43
Antibacterial activity of Surgihoney The antibacterial activity of Surgihoney (SH) against Staphylococcus aureus (NCIMB 9518) and the antibacterial activity of two prototype modified honeys made by Apis mellifera (honey bees) were tested. We also examined several modified forms of Surgihoney for their ability to change the level of hydrogen peroxide generation from the sample.

Methods: Surgihoney (SH) was compared to two modified honeys, prototype 1 (PT1) and prototype 2 (PT2) using a bioassay method against a Staphylococcus aureus standard strain. Further work considered the rate of hydrogen peroxide evolution from these formulations.

  Results: It was shown that the antibacterial activity of Surgihoney is mainly due to the generation of hydrogen peroxide. The modification of Surgihoney has been shown to produce two more potent honey prototypes, with antibacterial activity as high as between 2 and 3 times, and up to 10 times higher peroxide activity.

  Conclusion: Surgihoney is a clinically available wound antiseptic dressing that exhibits good antimicrobial activity. Two additional honey prototypes have been shown to have antibacterial activity that can be enhanced due to the demonstrated increase in peroxide activity.

Method 1. Measurement of Honey Activity by Bioassay Method The antibacterial activity of Surgihoney (SH) and two modified honeys, Prototype 1 (PT1) and Prototype 2 (PT2) was measured using Staphylococcus aureus (NCIMB 9518), Shown as% phenol equivalent. Values are calculated from average values from replicates of 3 samples repeated at 3 days.

Assay method. The agar well diffusion method used was Microbiology Standard Methods Manual for the New in the Year of the World and the World of the Worlds and the World of the Worlds.
In. New Zealand: Bee Products Standards
Council; 1982. From the punch plate assay for inhibitors described in

  Inoculum preparation. Sterile nutrient broth was used as a blank and diluent and the overnight culture was adjusted using a cuvette with a 1 cm optical path so that the absorbance measured at 540 nm was 0.5.

  Assay plate preparation. Using 100 μl of the culture adjusted to an absorbance of 0.5, seeded on 150 ml of nutrient agar to prepare an assay plate. The agar was spun to mix well, poured into a large petri dish and placed on a horizontal surface. As soon as the agar solidified, the plate was turned upside down and placed overnight and then used the next day. In the assay, these seeded plates were removed from 4 ° C. and allowed to stand at room temperature for 15 minutes, after which 7.0 mm diameter wells were cut on the agar surface. 250 μl of test material (sample or standard) was placed in each well.

  Catalase solution. A fresh solution of catalase 200 mg / ml from bovine liver (Sigma C9322, 2900 units / mg) in distilled water was prepared daily.

  Sample preparation. In a universal manner, 4 g of sample was added to 4 ml of distilled water to prepare the initial sample solution and placed at 37 ° C. for 30 minutes to facilitate mixing. To prepare the second solution, 2 ml of the first sample solution is universally added to 2 ml of distilled water and mixed for the total activity test, and 2 ml of the first sample solution is added to 2 ml of the catalase solution, Mixed for oxide activity.

  Preparation of phenol standards. Standard (w / v) 10%, 30% and 50% phenols were prepared by dissolving phenol in water. Phenol standards were brought to room temperature in the dark before use and mixed well before addition to test wells. Each standard was placed in 3 wells and tested in triplicate. The standard was stored at 4 ° C. and the expiration date was 1 month.

  Use of samples and standards. All samples and standards were tested in triplicate by adding 250 μl to each of the 3 wells.

  Plate incubation. After use of the sample, the plate was incubated at 37 ° C. for about 18 hours. The diameter of the blocking circle including the well diameter (7.0 mm) was recorded.

  Calculation of antibacterial activity of the sample. The mean diameter of a clear circle around each phenol standard was calculated and squared. A calibration diagram of% phenol against the square of the mean diameter of a clear circle was plotted. An optimized straight line was obtained using linear regression. Using this linear equation, the activity of each dilution of the honey sample was calculated from the square of the mean measurement of the clear circle diameter. This figure was multiplied by 4.69 to allow dilution (assuming that the density of Surgihoney is 1.35 g / ml), and the activity of the sample was expressed as equivalent to the phenol concentration (% w / v).

Total activity: Total activity including activity due to hydrogen peroxide (H ₂ O ₂ ).

Non-peroxide activity: Remove H ₂ O ₂ by treating the sample with catalase enzyme.

2. Measurement of honey activity by the H ₂ O ₂ method Mercoquant (registered trademark) 1.10011. And 1.10081 were used to measure activity.

The hydrogen peroxide test kit concentration was expressed as mg / L H ₂ O ₂ equivalent.

The sample was diluted 1:10 with purified water. After 5 minutes of incubation, all samples were measured over 12 hours for H ₂ O ₂ generation per hour, followed by 24 and 48 hour time points.

  Measuring method. Peroxidase carries oxygen from the peroxide to the organic redox indicator, which is then converted to a blue oxidation product. Peroxide concentration is measured semi-quantitatively by visually comparing the reaction zone of the test strip with the area of the color scale. The reaction zone of the test strip is immersed in the Surgihony sample for 1 second and excess liquid is allowed to flow from the strip onto the absorbent paper towel and after 15 seconds (Catalog No. 110011) or 5 seconds (Catalog No. 110081) of the color scale. Measure the color created in the reaction zone that more closely matches the area.

Result 1. Activity Evaluation Regarding the antibacterial activity produced by denaturation of the honey sample, the phenol activity in PT1 and PT2 was increased by 2 times and almost 3 times, respectively, as compared to Surgihoney alone. The results for three samples of Surgihoney (SH) and two modified prototypes, PT1 and PT2, are shown in Table 17.

2. Measurement of Honey Activity by the H ₂ O ₂ Method Prototype denaturation is observed to produce hydrogen peroxide activity that reaches 7 and 10 times that of Surgihoney. The results for three samples are shown in FIG. A linear relationship is observed by taking the maximum level of hydrogen peroxide generation for each of the three honey prototypes and plotting this against total phenol activity (FIG. 39).

Discussion The results from this work indicate that the main antimicrobial activity of Surgihoney (SH) and the two modified prototypes, PT1 and PT2, is due to hydrogen peroxide. This is a finding similar to other specific honeys from various flower sources. However, unlike previous work, the availability of hydrogen peroxide from the sample can be enhanced, and at 12 hours is 7 and 10 times the value of Surgihoney (SH) alone, respectively. There is a significant linear relationship between antibacterial activity and the maximum amount of hydrogen peroxide released from the three honey prototypes.

This peroxide activity provides a strong antimicrobial activity that is ideally suited for wound dressings used for acute or chronic wounds to treat or prevent wound infections. There is a small amount of catalase in the wound and it has been reported that the serum concentration of catalase in men is 50 kU / l, but the catalase activity in the healing wound is the first week after wounding. In fact, it has been shown that the activity level of catalase is restored to the initial level 2 weeks after wounding. It is therefore very unlikely that such catalase concentration will affect the antibacterial activity observed with Surgihoney or the two modified prototypes, PT1 and PT2, used on the outside.

  Ideal properties for antimicrobial wound dressings are efficacy, non-toxicity, ease of use, patient and clinician acceptance, and value for money. Hydrogen peroxide is an effective antibacterial agent and is already used as a biocide due to its strong activity against vegetable bacteria, yeast and spores. Hydrogen peroxide produces an antibacterial effect by chemical oxidation of cellular components.

  The toxicity of hydrogen peroxide to humans depends on concentration, and one study claims that specific concentrations for antibacterials and specific concentrations for human toxicity can overlap. In contrast, certain honey formulations are effective dressings by providing a low concentration of hydrogen peroxide to the wound continuously over time rather than supplying a large amount of hydrogen peroxide at once. It is an antibacterial agent and has been shown not to have such toxicity. Indeed, there is convincing evidence that providing physiological concentrations of hydrogen peroxide to mammalian cells stimulates biological responses and activates certain biochemical pathways in these cells.

  Clearly, Surgihoney and the two modified prototypes, PT1 and PT2, are antimicrobial coatings that provide effective hydrogen peroxide release over 24 hours.

Conclusion Surgihoney and the two modified prototypes, PT1 and PT2, have been shown to have potent antibacterial activity against Staphylococcus aureus standard strains. These antibacterial activities have been shown to be attributed to hydrogen peroxide. This activity can vary in magnitude and can be explained by the hydrogen peroxide activity. These modified honeys provide a coating that is effective, non-toxic and easy to administer.

Example 44
In this example, a preferred honey-based composition of the present invention is presented. The composition includes honey and added glucose oxidase (referred to as “active honey”).

Alcoholic gel Carbopol 940 0.3%
Triethanolamine 0.4% (required for pH adjustment and stability adjustment)
Active honey (5+) 65%
Ethanol 25.0%
Sufficient water Usage: Acne treatment. Do not use on damaged skin.

  “Active honey (5+)” is used to indicate that more glucose oxidase is used than other formulations without the (5+) designation.

Example 45
In this example, a preferred honey-based composition of the present invention is presented. The composition includes honey and added glucose oxidase (referred to as “active honey”).

Aqueous spray 25 ml plastic bottle contains:
Active honey 10g
Triton CF 0.1g
1 g maltodextrin or corn starch
Usage: Use for general wounds and burns

  Triton CF acts as a surfactant in this composition. Maltodextrin or corn starch acts as a film former.

Example 46
In this example, a preferred honey-based composition of the present invention is presented. The composition includes honey and added glucose oxidase (referred to as “active honey”).

Non-aqueous spray active honey 70%
30% propylene glycol
Usage: General wound

Example 47
In this example, a preferred honey-based composition of the present invention is presented. The composition includes honey and added glucose oxidase (referred to as “active honey”).

Coating material-Plastic pouch Various calcium alginate coating materials 10cm x 10cm, 20cm x 20cm
Active honey 10% w / w
Usage: Wound cleaning, wounds with relatively high exudation. This composition provides a relatively moist environment.

Example 48
In this example, a preferred honey-based composition of the present invention is presented. The composition includes honey and added glucose oxidase (referred to as “active honey”).

Honey and analgesics Active honey 99%
Ibuprofen 1%
Usage: Addresses all stages of wounds and provides pain management in wound healing

Example 49
In this example, a preferred honey-based composition of the present invention is presented. The composition includes honey and added glucose oxidase (referred to as “active honey”).

Powder active honey maltodextrin ibuprofen microcrystalline cellulose (CMC)
Polyvinylpyrrolidone (PVP)
Preferably, ibuprofen is included in the composition at 1-2% w / w. Other components can vary, for example, to obtain various moisture absorption levels.
Usage: Wrap in foil to prevent water absorption. Provides PVP-granulation. CMC-provides moisture drying power.

Example 50
In this example, a preferred honey-based composition of the present invention is presented. The composition includes honey and added glucose oxidase (referred to as “active honey”).

Wound cleaning Active honey (e.g., active honey 5-50%), optionally 1-2%
A reconstituted aqueous solution containing w / w analgesic, preferably also ibuprofen.
Usage: For example, placed in a dispenser (such as a pump dispenser) and used as a wash-off solution for wounds after debridement to provide early onset of wound healing and pain relief.

Example 51
In this example, a preferred honey-based composition of the present invention is presented. The composition includes honey and added glucose oxidase (referred to as “active honey”).

Foam covering Polyurethane backing for exudate absorption Insular part of calcium alginate soaked in active honey Silicon for preventing adhesion to wound Usage: Foam dressing that assists in sucking up wound exudate.

Example 52
In this example, a preferred honey-based composition of the present invention is presented. The composition includes honey and added glucose oxidase (referred to as “active honey”).

Treatment of onychomycosis A plaster containing active honey (5+) in the foam cavity Hydroxypropylcellulose Glycerol Isopropyl alcohol Citric acid monohydrate Usage: For treatment of fungal nail infection

Example 53
In this example, a preferred honey-based composition of the present invention is presented. The composition includes honey and added glucose oxidase (referred to as “active honey”).

Throat spray Contains 5-20% honey and glycerol, preferably honey. Usage: Sore throat. The formulation can be placed in a spray dispenser or reconstituted with water prior to use and then sprayed.

Example 54
In this example, a preferred honey-based composition of the present invention is presented. The composition includes honey and added glucose oxidase (referred to as “active honey”).
Pomade active honey 25%
White petrolatum Light liquid paraffin Talc Kaolin Zinc oxide Use: For the treatment of infectious eczema.

Example 55
In this example, a preferred honey-based composition of the present invention is presented. The composition includes honey and added glucose oxidase (referred to as “active honey”).

Lip Balm Petrolactam 5594 50%
Microcrystalline wax 9%
Cyclomethicone D5 31%
10% active honey
Usage: For cracking lips

Example 56
In this example, a preferred honey-based composition of the present invention is presented. The composition includes honey and added glucose oxidase (referred to as “active honey”).

Cream honey 15%
Carbomer 2.63%
Dimethicone 0.13%
Sodium lauryl sulfosuccinate 0.05%
Edetate disodium 0.13%
Glycerol 5.26%
Colloidal silicate 0.33%
Poloxamer 0.26%
Sodium hydroxide 0.41%
Purified water 85.03%
Usage: for acne treatment. Honey is known to be active against Propionibacterium acnes.

Claims (1)

  The invention described herein.