JP2020063234A - Antimicrobial compositions - Google Patents

Abstract

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 to treat microbial infections and wounds is described, as well as methods for their production.SELECTED DRAWING: None

Description

The present invention relates to compositions for the generation of antibacterial activity, in particular storage-stable compositions, antibacterial compositions and solutions, the use of these compositions and solutions, especially for wound healing, and the process for their preparation.

Honey has been used since ancient times to treat microbial infections. In recent years, there has been a renewed interest in the therapeutic effects of honey, especially in the field of wound healing. In clinical trials, honey is an effective broad-spectrum antibacterial agent, and Pseudomonas aeruginosa, Staphyloc
occus aureus, Candida albicans and Escherich
It has been shown to be effective against common microorganisms that infect wounds, such as ia coli, and against antibiotic resistant strains of bacteria. 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, p
Due to H, hydrogen peroxide production and the presence of phytochemical constituents. Manuka Tree (Leptospe)
Manuka honey produced from rumum scoparium) is recognized as having superior antibacterial activity compared to most other honeys due to the high level of antibacterial phytochemical activity present in this type of honey. There is. However, Manuka honey has a relatively limited supply and cannot meet the demand in the world market.

In honey, in addition to its antibacterial action, several other actions are believed to aid in wound healing, especially in chronic wounds. In particular, honey has the ability to autolytically debride and deodorize wounds. Debridement is the removal of dead, damaged or infected tissue,
It is to improve the healing ability of the remaining healthy tissue. Honey has also been reported to have anti-inflammatory properties, capable of stimulating tissue growth, and coping with pain to minimize scarring.

Several honey based wound care dressings are currently commercially available. Many of these use manuka honey or other honeys that have high levels of non-peroxide antibacterial activity. Because, acidity, catalase activity, and protein-digesting enzymes present in wound exudates can reduce the antimicrobial effect of hydrogen peroxide.

Currently available honey-based coating materials include Comvita / Derma Sci
medianesie product line containing manuka honey manufactured by Ences, and Actu, an additive-free pure manuka honey manufactured by Advancis.
Ivon is mentioned. Other honey-based dressings contain honey and one or more additional components. Examples of these include Melladerm Plus, an antimicrobial wound gel manufactured by SanoMed. This gel contains honey from the Bulgarian multifloral mountain region, which naturally has a high content of glucose oxidase and phenol (the main phytochemical constituents in honey are phenolic compounds). . During processing, the honey is not heated or irradiated. This is because it is believed that it eliminates the healing properties, especially the production of hydrogen peroxide by glucose oxidase. Instead, it is sterilized using the ozone treatment method described in WO 2008/049578. In this method, ozone gas is bubbled into the liquefied honey in an ozone resistant container. However, ozone is only approved by the US Food and Drug Administration (FDA) as a disinfectant for reusable medical devices, and is therefore currently approved by the US FDA for disinfecting honey-based products for wound healing applications. It has not been.

The Mesitran family is a further family of honey-rich wound care products. T
L-Mesitran ointment manufactured by Ritticum contains 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. Contains.

The difference in antibacterial activity between honeys can be greater than 100-fold depending on the geographical, seasonal and botanical sources of honey, as well as harvesting, processing and storage conditions. Therefore, the honey-based coating material has different antibacterial effects depending on the type of honey used. Research by Jenkins, Burton and Cooper ("The dete
termination of antimicrobial activity of
three honey impregnated wound dressings b
y challenge test with EMRSA-15 ", Advances
), Honey-based dressings containing Activon, Medihoney or Mesitran products were all found to have different efficacy against EMRSA-15. The dressing impregnated with Activon was the most effective, followed by the Medihoney coating, which was much less effective with the Mesitran product.

Therefore, there is a need to reduce the variability in the antibacterial activity of honey and improve the antibacterial activity of honey with low antibacterial activity. It is also desirable to provide effective honey-based wound care products that have high levels of phytochemicals regardless of the use of honey. It is also desirable to provide an effective honey wound care product that has not been 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 (R) Ag dressing, which provides controlled silver ion formation when wound exudate is absorbed into the dressing. Release with. However, Staphylococcus aureus, P
Organisms resistant to silver, such as Seudomonas aeruginosa and enterococci, have been reported and may delay wound healing.

Povidone iodine (PVP-I) is a stable chemical complex of polyvinylpyrrolidone (povidone, PVP) and elemental iodine. It is a broad spectrum antiseptic for topical use in wound healing 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 about its safety and efficacy, and its use in pressure ulcers was not recommended by the US Department of Health and Human Services.

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

Therefore, there is also a need to provide a broad spectrum active antimicrobial wound care product that is non-toxic and can be used in the treatment of chronic wounds.

International Publication No. 2008/049578

Jenkins RE, Burton NF, Cooper RA, "The termination of antimicrobial activation of three honey imprinted wounds 15th anniversary by RSVP".

Applicants have recognized that the antibacterial activity of honey depends on a delicate interrelationship between the natural inhibitor and the active agent, with low variability between different types and different collections of honey. We are developing a composition capable of exhibiting the above antibacterial activity and improving the antibacterial properties of honey having poor antibacterial activity. Applicants have also found that such compositions have excellent wound healing properties even after sterilization by gamma irradiation exposure.
Applicants recognize that these findings have applicability to other natural substances as well.

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

Broadly, the invention provides compositions for generating antimicrobial activity. The composition comprises an enzyme capable of converting a substrate to release hydrogen peroxide, and a substance containing a substrate for the enzyme. This enzyme refers to any enzyme activity that may be present in the above substances and is capable of converting a substrate to release hydrogen peroxide (referred to herein as “substrate converting activity”). It is additive (ie, added as a result of human intervention). Preferably the composition is
A storage-stable composition that does not contain sufficient 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, which comprises a purified enzyme capable of converting a substrate to release hydrogen peroxide; The composition does not include sufficient free water for the enzyme to convert the substrate.

The present invention also provides a method for producing a storage-stable composition for generating antibacterial activity, which comprises purifying a purified enzyme capable of converting a substrate to release hydrogen peroxide to the enzyme. Including contacting with a substrate-containing substance, the composition does not contain enough free water for the enzyme to convert the substrate.

According to 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 substance lacking 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 comprises converting an enzyme capable of converting a substrate to release hydrogen peroxide, and a substrate for the enzyme. And contacting a substance lacking catalase activity, the composition does not contain enough free water for the enzyme to convert the substrate.

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

According to the present invention, there is also provided a method of making an antimicrobial composition or solution, the method comprising the step of: providing a storage-stable composition of the invention sufficient for an enzyme to convert a substrate to release hydrogen peroxide. Includes diluting with fresh water.

According to the present invention there is further provided an antimicrobial composition or solution, which composition or solution comprises:
It comprises a storage-stable composition of the present invention diluted with sufficient water to release hydrogen peroxide by enzymatic conversion of the substrate.
For example, the present invention provides the following items.
(Item 1)
A storage-stable composition for generating antibacterial activity, which comprises a purified enzyme capable of converting a substrate to release hydrogen peroxide; and an unpurified natural substance containing a substrate for the enzyme;
Wherein the composition does not contain sufficient free water for the enzyme to convert the substrate.
(Item 2)
A storage-stable composition for generating antibacterial activity, wherein the composition can convert a substrate to release hydrogen peroxide; and an unpurified composition having no catalase activity and containing a substrate for the enzyme Natural substances;
Wherein said enzyme is additive to any enzyme activity that may be present in said unpurified natural substance and which is capable of converting said substrate to release hydrogen peroxide, said composition Wherein the composition does not contain sufficient free water for the enzyme to convert the substrate.
(Item 3)
Item 2. The composition according to Item 1, wherein the unpurified natural substance has no catalase activity.
(Item 4)
4. The composition according to item 2 or 3, wherein the substance has been treated to remove or inactivate catalase activity.
(Item 5)
The composition according to any of the preceding items, wherein said substance is a crude 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 the catalase activity or has been inactivated.
(Item 8)
Item 6. The composition according to Item 5, wherein the sugar substance is syrup or inversion syrup.
(Item 9)
The composition according to any one of the preceding items, wherein the substance has no enzymatic activity capable of converting the substrate to release hydrogen peroxide.
(Item 10)
The composition according to any of the preceding items, wherein said substance is a pasteurized substance.
(Item 11)
Item 7. The composition according to Item 6, wherein the honey is honey that has not been pasteurized, and preferably creamy honey that has not been pasteurized.
(Item 12)
The composition according to any of the preceding items, wherein said enzyme is an oxidoreductase enzyme.
(Item 13)
The oxidoreductase enzyme is glucose oxidase, hexose oxidase,
13. The composition according to item 12, which is 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 according to item 15, wherein the composition contains 1 unit or more of glucose oxidase per gram of the composition.
(Item 17)
17. The composition of 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)
The composition contains 100 units or more, preferably 100 to 5 per gram of the composition.
19. A composition according to any of items 16-18, comprising 00 units of glucose oxidase.
(Item 20)
The composition contains 500 units or more, preferably 500 to 1 per gram of the composition.
20. The composition according to any of items 16-19, which comprises 000 units of glucose oxidase.
(Item 21)
The composition according to any of the preceding items, wherein the composition is a food grade composition.
(Item 22)
The composition according to any of the preceding items, wherein said composition is a medical grade or medical device grade composition.
(Item 23)
The composition according to any of the preceding items, wherein each component of the composition is a natural substance.
(Item 24)
The composition according to any of the preceding items, wherein said composition is in powder form.
(Item 25)
25. An antibacterial composition comprising the composition according to any of items 1-24 diluted with water sufficient 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 converting a substrate in the unpurified natural substance to release hydrogen peroxide. .
(Item 27)
A composition for generating antibacterial activity, the composition comprising an unpurified natural substance having no catalase activity; and an enzyme capable of converting a substrate in the unpurified natural substance to release hydrogen peroxide. The composition comprising, wherein the enzyme is in addition to any enzymatic activity capable of converting the substrate to release hydrogen peroxide, which may be present in the crude natural substance.
(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 of items 26-28, wherein the composition further comprises hydrogen peroxide.
(Item 30)
The composition according to any of the preceding items, wherein said composition is a sterile composition.
(Item 31)
31. The composition of item 30, wherein the composition has been sterilized by gamma irradiation exposure.
(Item 32)
An antibacterial solution comprising the composition according to any of items 26 to 31.
(Item 33)
A method for sterilizing the composition according to any one of Items 1 to 29, wherein the method is preferably 10 to 70 kGy, more preferably 25 to 70 kGy, and most preferably 35 to 70 k.
The method comprising exposing the composition to gamma gamma irradiation of Gy.
(Item 34)
A method for producing a storage-stable composition for generating antibacterial activity, the method comprising a purified enzyme capable of converting a substrate to release hydrogen peroxide, and an unpurified natural substance containing a substrate for the enzyme. Wherein the composition does not contain sufficient free water for the enzyme to convert the substrate.
(Item 35)
A method for producing a storage-stable composition for generating antibacterial activity, the method comprising an enzyme capable of converting a substrate to release hydrogen peroxide, the substrate containing the enzyme, and having no catalase activity. The method, comprising contacting with unpurified natural material, wherein the composition does not contain sufficient free water for the enzyme to convert the substrate.
(Item 36)
The method according to Item 34, wherein the unpurified natural substance has no catalase activity.
(Item 37)
37. The method according to any of items 34-36, wherein the substance is a crude natural sugar substance.
(Item 38)
Any substance present in the substance capable of treating the substance prior to contacting the substance with the enzyme to convert catalase activity present in the substance and / or converting the substrate to release hydrogen peroxide. 38. The method according to any of items 34 to 37, wherein the enzymatic 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 eliminate 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 converting activity and then cooling the substance to a temperature at which the enzyme can maintain activity before contacting the substance with the enzyme. The method according to item 39.
(Item 41)
41. The method according to any of items 34-40, wherein the substance is honey.
(Item 42)
38. The method according to item 37, wherein the unpurified natural sugar substance is syrup or inversion syrup.
(Item 43)
43. The method of paragraph 42, wherein the method further comprises converting a substance comprising sucrose to form the converted syrup.
(Item 44)
44. The method according to any of items 34-43, wherein the enzyme is an oxidoreductase enzyme.
(Item 45)
The oxidoreductase enzyme is glucose oxidase, hexose oxidase,
45. The method according to item 44, which is cholesterol oxidase, galactose oxidase, pyranose oxidase, choline oxidase, pyruvate oxidase, glycolate oxidase or amino acid oxidase.
(Item 46)
46. The method according to 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 of items 34 to 47 for producing a food grade composition, wherein the substance and the enzyme are food grade.
(Item 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 to release hydrogen peroxide with an unpurified natural substance containing a substrate for the enzyme. The method comprising:
(Item 51)
A method for producing a composition for generating antibacterial activity, the method comprising an enzyme capable of converting a substrate to release hydrogen peroxide, a crude natural substance containing a substrate for the enzyme and having no catalase activity. Said method comprising contacting with.
(Item 52)
The method according to Item 50, wherein the unpurified natural substance has no catalase activity.
(Item 53)
53. The method according to any of items 34-52, wherein the enzyme and the unpurified natural substance are in powder form.
(Item 54)
A method for producing an antibacterial solution, wherein the method converts the storage-stable antibacterial composition according to any one of Items 1 to 24 by converting the substrate in the sugar substance by the enzyme to hydrogen peroxide. Diluting with sufficient water to release the.
(Item 55)
The composition according to any one of Items 1 to 31 or the antibacterial solution according to Item 32 of use as a pharmaceutical.
(Item 56)
33. The composition according to any one of Items 1 to 31 or the antibacterial solution according to Item 32 for preventing, treating or ameliorating microbial infection.
(Item 57)
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 preventing, treating, or improving microbial infection.
(Item 58)
A method for preventing, treating or ameliorating a microbial infection, wherein the method requires the composition according to any one of Items 1 to 31 or the antibacterial solution according to Item 32 for such prevention, treatment or improvement. The method comprising administering to a subject.
(Item 59)
59. The method of item 58, wherein said method comprises topically administering said composition or said solution.
(Item 60)
60. The use or method according to any of items 55-59 for treating a human or animal subject.
(Item 61)
38. A method for treating a wound, the method comprising administering to the wound site the composition according to any of items 1 to 31 or the antimicrobial solution according to item 32.
(Item 62)
33. The composition according to any one of items 1 to 31 or the antibacterial solution according to item 32 for treating a wound.
(Item 63)
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 a wound.
(Item 64)
A method for treating inflammation, which comprises administering the composition according to any one of Items 1 to 31 or the antibacterial solution according to Item 32 to the site of inflammation.
(Item 65)
33. The composition according to any one of Items 1 to 31 or the antibacterial solution according to Item 32 for treating inflammation.
(Item 66)
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, the method comprising administering the composition according to any of items 1 to 31 or the antibacterial solution according to item 32 to a site in need of such stimulation. The 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)
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, said 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 the antibacterial solution according to Item 32 for debriding.
(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 debriding.
(Item 73)
A method for deodorizing a wound, the method comprising administering the composition according to any one of Items 1 to 31 or the antibacterial solution according to Item 32 to a wound in need of deodorization.
(Item 74)
33. The composition according to any one of items 1 to 31 or the antibacterial solution according to item 32 for deodorizing a wound.
(Item 75)
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 deodorization.
(Item 76)
59. The method according to item 58, wherein the object is an insect.
(Item 77)
77. The method according to item 76, wherein the insect is a bee.
(Item 78)
The method according to Item 77, for preventing, treating or ameliorating Nosema infection.
(Item 79)
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. A 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)
An antibacterial composition adapted for topical administration to a subject, wherein the antibacterial composition comprises the composition according to any one of Items 1 to 31 or the antibacterial solution according to Item 32.
(Item 84)
An antibacterial composition adapted for oral administration to a subject, wherein the antibacterial composition comprises the composition according to any one of Items 1 to 31 or the antibacterial solution according to Item 32.
(Item 85)
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 of preventing or inhibiting microbial growth, said method preventing or inhibiting such growth of a composition according to any of items 1-31 or an antimicrobial solution according to item 32. Using the desired surface or area.
(Item 87)
87. A method according to item 86, wherein the method comprises using the composition or the solution to prevent or prevent the growth of plant or entomopathogenic fungi.
(Item 88)
Said method uses said composition or said solution to prevent the growth of Erwinia,
89. A method according to item 86 or 87, which comprises blocking.
(Item 89)
89. The method according to any of items 86-88, wherein the method comprises using the composition or the solution on a plant, fruit or vegetable.
(Item 90)
90. The method according to item 89, wherein the vegetable is potato.
(Item 91)
Use of the composition according to any of items 1 to 31 or the antibacterial solution according to item 32 for preventing or inhibiting microbial growth.

Figure 5 shows the results of a well diffusion assay for the antibacterial effect of pasteurized Ulmo honey spiked with different amounts of glucose oxidase. 7 shows the results of a well diffusion assay for the antibacterial effect of compositions of the present invention with different amounts of enzyme compared to the effect of different phenol standards. Figure 3 shows a photograph of the results of a well diffusion assay for the antibacterial effect of pasteurized Tineo honey supplemented with glucose oxidase. 7 shows the results of a well diffusion assay for the antibacterial effect of different compositions of the invention. 7 shows the results of a well diffusion assay for the antibacterial effect of compositions of the invention containing different enzyme formulations. Figure 5 shows the results of a well diffusion assay for antibacterial efficacy of compositions of the invention after storage for 60 or 90 days. 5 shows the results of a well diffusion assay for the antibacterial effect of powder formulations of the composition of the invention. 5 shows the results of a well diffusion assay for the antibacterial effect of various products in the presence and absence of the composition of the invention. 5 shows the results of a well diffusion assay for the antibacterial effect of various products in the presence and absence of the composition of the invention. 5 shows the results of a well diffusion assay for the antibacterial effect of various products in the presence and absence of the composition of the invention. 5 shows the results of a well diffusion assay for the antibacterial effect of various products in the presence and absence of the composition of the invention. Figure 5 shows the results of a well diffusion assay for the antimicrobial effect of the sterile honey-based composition of the present invention compared to Manuka UMF25 + Honey. The therapeutic effect of an infected toe using the sterile honey type composition of the present invention is shown. The therapeutic effect of a toe ulcer using the aseptic honey type composition of this invention is shown. The therapeutic effect of a toe ulcer using the aseptic honey type composition of this invention is shown. The therapeutic effect of a foot ulcer using the aseptic honey type composition of the present invention is shown. The therapeutic effect of a foot ulcer using the aseptic honey type composition of the present invention is shown. The therapeutic effect of a foot ulcer using the aseptic honey type composition of the present invention is shown. The therapeutic effect of a foot ulcer using the aseptic honey type composition of the present invention is shown. The therapeutic effect of a traumatic leg wound using the sterile honey-based composition of the present invention is shown. The therapeutic effect of an infectious leg wound using the sterile honey-based composition of the present invention is shown. The therapeutic effect of a foot ulcer using the aseptic honey type composition of the present invention is shown. The therapeutic effect of leg ulcers using the sterile honey-based composition of the present invention is shown. The therapeutic effect of leg ulcers using the sterile honey-based composition of the present invention is shown. The therapeutic effect of leg ulcers using the sterile honey-based composition of the present invention is shown. The therapeutic effect of leg ulcers using the sterile honey-based composition of the present invention is shown. The therapeutic effect of leg ulcers using the sterile honey-based composition of the present invention is shown. The therapeutic effect of leg ulcers using the sterile honey-based composition of the present invention is shown. The therapeutic effect of leg ulcers using the sterile honey-based composition of the present invention is shown. The therapeutic effect of leg ulcers using the sterile honey-based composition of the present invention is shown. The therapeutic effect of leg ulcers using the sterile honey-based composition of the present invention is shown. 7 shows the therapeutic effect of diabetic leg ulcer associated with latent osteomyelitis using the sterile honey-based composition of the present invention. The therapeutic effect of the pressure sore which uses the aseptic honey type composition of this invention is shown. The therapeutic effect of infectious axillary wounds using the sterile honey-based composition of the present invention is shown. 5 shows the therapeutic effect of an infectious disease around the entrance of a catheter using the sterile honey-based composition of the present invention. Shown are the time kill curves of Surgihoney 1 (S1), Surgihoney 3 (S3), and Medihoney (MH) against different test organisms: (a) Staphylococcus aureus; (b) Methicillin resistant Staphylococcus SA. (d) Vancomycin-resistant enterococcus (VRE); (e) Pseudomonas aeruginosa; (f) Klebsiella; (g) E. coli. coli ESBL; (h) Enterococcus faecalis. Shown are the time kill curves of Surgihoney 1 (S1), Surgihoney 3 (S3), and Medihoney (MH) against different test organisms: (a) Staphylococcus aureus; (b) Methicillin resistant Staphylococcus SA. (d) Vancomycin-resistant enterococcus (VRE); (e) Pseudomonas aeruginosa; (f) Klebsiella; (g) E. coli. coli ESBL; (h) Enterococcus faecalis. Shown are the time kill curves of Surgihoney 1 (S1), Surgihoney 3 (S3), and Medihoney (MH) against different test organisms: (a) Staphylococcus aureus; (b) Methicillin resistant Staphylococcus SA. (d) Vancomycin-resistant enterococcus (VRE); (e) Pseudomonas aeruginosa; (f) Klebsiella; (g) E. coli. coli ESBL; (h) Enterococcus faecalis. 5 shows the results of plaque assay testing the antiviral activity of S1 and S2 Surgihoney. 5 shows photographs of the results of using S1 Surgihoney to treat 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. 5 shows photographs of the results of using S1 Surgihoney to treat pressure ulcers. (A) shows the 1st day of treatment; and (b) shows the 30th day of treatment. Figure 2 shows the different hydrogen peroxide production rates of Surgihoney (SH) and two modified prototypes, PT1 and PT2. Figure 3 shows the relationship between phenolic activity and maximal 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. The use of substances without catalase activity means that there is no variation in the amount of this activity between similar substances from different sources, or similar substances from different harvests from the same source. This reduces the variability in antibacterial activity that can occur from such materials. Alternatively, if the substance contains catalase activity and it is not possible or desirable to inactivate the catalase activity in the substance prior to contacting the substance with an enzyme, then the peroxidation that can be generated from the substance. Enough enzyme may be used to reduce the effect of catalase activity on hydrogen. This also reduces the variation in antibacterial activity that can occur from the substances mentioned above. 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-listed material, or it may be inactivated prior to its use in the compositions of the present invention. Catalase activity may be heat inactivated, for example by pasteurization. Suitable temperature for heat inactivation of catalase activity is at least 60 ° C, 70 ° C or 80 ° C, preferably for 2 minutes or more.

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 of the composition to generate antibacterial activity after dilution. More preferably, it is used to mean that it can be stored for 1 or 2 months or more. A 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. The lack of sufficient free water in the storage-stable composition of the present invention prevents the enzyme from converting the substrate to release hydrogen peroxide, thus improving the stability of the composition at room temperature for extended periods of time. Help maintain. The storage-stable compositions of the present invention may include some water, provided there is not enough free water for the enzyme to convert the substrate. The suitable amount of water will vary depending on the specific components of the composition. However, typically, the storage stable composition of the present invention preferably has a total water content of less than 20%,
For example, it contains 10% to 19% water.

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

The enzyme present in the composition of the present invention may be any enzyme activity which may be present in the above substances and is capable of converting a substrate to release hydrogen peroxide (herein, “substrate converting activity”). "
Referred to above), that is, the composition of the present invention comprises the above-mentioned substance and the added enzyme. In some embodiments, it is preferred that the above substances have no substrate conversion activity.

It is understood that after dilution of the storage-stable composition of the present invention, sufficient enzyme should be present in the composition to convert the above substrates to form hydrogen peroxide as needed. See.

Since 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. The term "purified enzyme" is used herein to include an enzyme preparation in which the enzyme is separated from at least some of the impurities that were originally present when the enzyme was made. Preferably, the impurities that have been removed or reduced include those that would otherwise interfere with the ability of the enzyme to convert the substrate to release hydrogen peroxide.

It is not necessary or desirable for the purified enzyme to be at a high level of purity, as long as the enzyme is capable of converting the substrate to release hydrogen peroxide. May be. In some situations, it may be desirable to use a relatively crude enzyme formulation. Examples of suitable purity levels include at least 10%, 2
Examples include 0%, 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 beginning when making the enzyme is reduced. This enzyme may be produced by recombinant means or non-recombinant means, and may be recombinant enzyme or non-recombinant enzyme. The enzyme may be purified from a microbial source, preferably a non-genetically modified microorganism.

Purity levels of the above enzymes may be selected as needed depending on the intended use of the above composition. For example, food grade purity may be appropriate where the composition is intended for human consumption. Medical applications require the use of medical grade or medical device grade purity.

Preferably the enzyme is an oxidoreductase enzyme. Examples of oxidoreductase enzymes capable of converting a 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. Substrates corresponding to these oxidoreductase enzymes are D-glucose, hexose, cholesterol, D-galactose, pyranose, choline, pyruvic acid, glycolic acid and amino acids, respectively.

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

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

The substance may be any substance containing a substrate for the enzyme. Preferably this substance is devoid of catalase activity. Preferably this material is a crude material. The term "unpurified" is used herein to refer to material that has not been processed into its pure form. Unrefined substances include substances which may have been concentrated, for example by drying or boiling.

Preferably, the above substances include one or more substrates from natural sources (referred to herein as "natural substances"). Examples of natural substances include substances from botanical sources, including substances from sap, roots, nectar, flowers, seeds, fruits, leaves or shoots. More preferably this material is a crude natural material.

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

Preferably the substance is a sugar substance. The term "sugar substance" is used herein to mean any substance containing one or more sugars. As used herein, "sugar"
The term 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 substance comprises one or more sugars from natural sources (referred to herein as "natural sugar substances"). More preferably, the natural sugar substance is a crude natural sugar substance.
The crude natural sugar material may be (or may be derived from) a natural sugar product. In a preferred embodiment, the crude natural sugar product is honey. In some preferred embodiments, the honey is honey that has been treated to remove or deactivate catalase activity. Alternatively, this unpurified natural sugar substance is
It may be a processed natural sugar such as a syrup or invert syrup.

As explained above, preferably the substance itself may not have an enzymatic activity capable of converting the substrate to release hydrogen peroxide (referred to as “substrate converting activity”). The lack of substrate-converting 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 different substances from the same source. Have. This further reduces the variation in antibacterial activity that can occur from such substances. If so, the substrate conversion activity is brought about only by the enzyme that comes into contact with the above substances,
And the amount of substrate converting activity present in the above composition can be controlled.

Substrate converting activity may be removed during processing or harvesting of the above material, or it may be inactivated prior to its use in the compositions of the present invention. Substrate conversion activity may be inactivated by heat inactivation, for example by pasteurization. A suitable temperature for thermal 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 include an antimicrobial agent. For example, the enzyme is contacted with the substance in an aqueous solution under conditions of substrate conversion by the enzyme, and then the composition is dried to free water that is not sufficient for the enzyme to convert the substrate. Hydrogen peroxide may be present when reducing the water content of the composition to a level where it is only present to form the above storage-stable composition. However, preferably, the storage stable antimicrobial composition described above does not contain detectable hydrogen peroxide. Such compositions 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 antibacterial agents that may be present in the storage stable compositions of the present invention include antibiotics, antiviral or antifungal agents.

It is particularly preferred that the composition of the present invention is a food grade composition. Such compositions may be used for human consumption. In another particularly preferred embodiment, the composition is a medical grade or medical device grade composition. Alternatively, the composition of the invention may be an animal food standard for animal consumption.

It is particularly preferred that each component of the above compositions 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 be preferably medical grade or medical device grade honey. In some embodiments, the honey is preferably honey that has been treated to remove or deactivate the 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 or medical device grade honey and the enzyme is a medical or medical device 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 Codex Alimenta
In the rius international food standard, "a natural sweet substance made by honey bees, which is a natural sweetener produced by bees from nectar of plants, secretions from living parts of plants, or discharges of insects sucking plants in living parts of plants, It was collected, changed by mixing with a special substance of honey bees, placed, dehydrated, stored in beehives, left to mature and mature ”(Revised Codex Standard ford). Honey,
2001).

Nectar normally contains about 14% monosaccharides (w / w), 1% phenolic compounds, and 85% water. This phenolic compound imparts flavor, aroma and color to the honey. In warm conditions in a honeycomb, usually 36 ° C, nectar ferments very quickly. To prevent this, nectar is mixed with secretions from the salivary and hypopharyngeal glands of foraged honeybees that contain the enzyme. In beehives, nectar is passed from bee to bee, adding more secretions before being stored in the honeycombs of beehives. The amount of enzyme present depends on the age of the bees, the diet and the physiological stage (bees produce more digestive enzymes in the glands when they feed on), colony strength, beehive temperature, and nectar flux. 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 gluconate. Glucose oxidase which catalyzes Low doses of hydrogen peroxide prevent the growth of yeast, which ferment nectar quickly. When bees progressively dry nectar to form honey, gluconic acid makes the honey acidic (pH 3.5 to 4.5). In honey, water is effectively trapped in sugar molecules and unavailable for further chemical reactions. The amount of "free" water in honey is measured as the water activity ( _aw ). The range of a _w found in honey is reported to be 0.47-0.70 with average values of 0.562 and 0.589 (RC
IEGG, M; BLANC, B, 1981, The water activ
ity of honey and related sugar solutions
． Lebensmittel-Wissenschaft und Technology
gie 14: 1-6). The _aw of aged honey is so low that growth of any species is not maintained and fermentation does not occur below a moisture content of 17.1% (Molan, P. et al.
C. (1992). The antibacterial activity o
f honey: 1. The nature of the antibacter
ial activity. Bee World, 73 (1), 5-28). The acidity of honey and the lack of free water prevent further fermentability and stop the action of glucose oxidase. Honey also contains an indeterminate amount of catalase originating from nectar.

The typical chemical composition of flower honey is as follows.

In addition, there are trace amounts of pollen and the enzymes invertase, diastase, catalase and glucose oxidase. Pollen can be used to identify the plant origin of honey. Phytochemical components are also present. This varies depending on the source of honey, but is typically about 1% or less.

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

According to embodiments of the present invention, the honey may be pasteurized. Pasteurization of honey 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 at which the added glucose oxidase is not inactivated, and this pasteurized honey is sufficient to facilitate mixing with glucose oxidase. After cooling the honey to a temperature at which it remains liquid (preferably 35 to 40 ° C.), the honey and glucose oxidase are contacted.

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

Controlling the heat treatment may be important, as a by-product of heating honey is the production of HMF (hydroxymethylfurfuralaldehyde), which is used as an indicator of thermal and storage changes in honey. HMF is produced by the decomposition of fructose in the presence of acid. Heat increases the rate of this reaction. The velocity increases exponentially with increasing heat. Every time the honey rises by 1 ° C above 40 ° C, which is close to the ambient temperature of normal honeycombs,
HMF increases rapidly. HMF is not a harmful product. Jam, molasses, golden syrup, etc. may have an HMF concentration that is 10 to 100 times that of honey. However, HMF concentration is used as an index of honey deterioration, and Codex Alime
According to Ntarius Standard, 40 mg / l is the maximum permissible concentration for table honey in EU.

To prevent an increase in HMF, the honey is rapidly warmed to a temperature level at which catalase is inactivated and then a heat exchange mechanism is used to raise the temperature of this honey to a maximum of 40-
Rapid reduction to 45 ° C is preferred.

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

  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,
The small crystals hinder the formation of large crystals that can occur in raw honey. A method of making creamy honey was described in US Pat. No. 1,987,893. In this processing, the raw material honey is first pasteurized, and then the creamy honey that has been processed in advance is added to this pasteurized honey to prepare a mixture of 10% creamy honey and 90% pasteurized honey. The mixture is then allowed to rest at a controlled temperature of 14 ° C. This method produces one batch of creamy honey in about a week. Seed batches can be made by crystallizing regular honey and crushing the crystals to the desired size. Large scale manufacturers modified this process by stirring the mixture using a stir bar while keeping the above honey mixture at 14 ° C. In an alternative creaming method, the pasteurization step described above 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 food grade for human consumption, or medical grade or medical device grade. The activity of glucose oxidase may be selected according to the desired rate of hydrogen peroxide evolution 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 Aspergillus n.
iger, Penicillium amagasakiense, Penicilli
um variabile and Penicillium notatum selected strains are mentioned. Medical device grade glucose oxidase from GMO Aspergillus niger is available from Biozyme UK and has an activity of 240 iu / m.
It is g. Food grade glucose oxidase from Aspergillus niger is available from BIO-CAT INC with 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 from Aspergillus niger (GO3B2) is available from BBI Enzymes Limited and has an activity of 360 units / mg. Contaminants: Alpha amylase 0.0
5% or less, saccharase 0.05% or less, maltase 0.05% or less, and GO / Cat
Over 2000.

The activity of the above enzyme (for example, glucose oxidase activity) is, 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 on the substance, the amount of substrate present on the substance, the desired level of hydrogen peroxide release, and the desired excess. It appears to be due to several factors, including the length of hydrogen oxide release time. Suitable enzyme amounts can be readily determined by one of skill in the art and, if necessary, in Example 2 below.
A well diffusion assay, as described in, can be used to measure and determine the extent of hydrogen peroxide release for different amounts of enzyme. A suitable amount of enzyme (such as glucose oxidase) may be 0.0001% to 0.5% w / w of the composition described above. The amount of enzyme used may be selected to produce a composition that produces an antimicrobial activity equal to the selected phenol standard (eg, 10%, 20% or 30% phenol standard).

Compositions of the invention, especially those in which the above substances are honey (eg, non-pasteurized honey)
The composition of the present invention, in which the enzyme is glucose oxidase capable of converting D-glucose in honey to release hydrogen peroxide, is 1 unit or more per 1 gram of the composition, preferably May contain up to 1500 units of glucose oxidase. This glucose oxidase is additive (ie added as a result of human intervention) to any glucose oxidase that may be naturally present in the above substances.

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

Applicants have found that the antibacterial potency 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 present invention, the composition of the present 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. One such composition is 1 gram of this composition.
It has been found to have superior antibacterial properties to compositions having 5 units or less of glucose oxidase. In particular, such compositions include MSSA, MRSA, Groups A and B
Streptococci, Enterococcus, E .; coli, E. coli
ESBL, Serr. liquefaciens Amp C, Kleb. pneum
oniae, Pseud. aeruginosa and Candida albicans
It has high efficacy against a wide range of microorganisms including.

In another embodiment 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 antibacterial properties. In particular, such compositions are disclosed in MSSA, MRSA, E.I. coli
It has even higher potency against a broad spectrum of microorganisms, including ESBL and Staphylococcus aureus.

The pasteurization process described above inactivates any enzyme activity present in the honey described above, so that catalase activity between pasteurized honeys from different sources, or between different harvests of honey from the same source. And there is no change in substrate conversion activity. The amount of substrate exchange activity can be controlled by the addition of a purified glucose oxidase formulation with a defined amount and a defined activity enzyme. Thus, the variability in the natural antibacterial activity between honeys of different types and different harvests is significantly reduced and the antibacterial properties of low antibacterial honeys 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. The term "conversion" is used herein to mean that the sucrose originally present in the substance has been converted to glucose and fructose, for example by the activity of the invertase or sucrase enzyme. Accordingly, such additive materials may be used in the compositions of the present invention having the glucose oxidase enzyme.

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

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

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

The present invention further provides an antibacterial composition containing a substance and a purified enzyme capable of converting a substrate in the substance to release hydrogen peroxide, or a composition for generating antibacterial activity.

According to the present invention, the production of an antibacterial composition or a composition for generating antibacterial activity, which comprises contacting a purified enzyme capable of converting a substrate to release hydrogen peroxide with a substance containing a substrate for this 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 antimicrobial solution comprising the composition or antimicrobial 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, mycobacteria, 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, Staphy
lococcus saprophytics, beta hemolytic Streptococci
Group A or B, Campylobacter coli, Campylobacter
er jejuni, Methicillin-resistant Staphylococcus Aureus (M
RSA), methicillin-sensitive Staphylococcus Aureus (MSSA)
, Botrytis cinerea, Mycobacterium tubercul
osis, Cryptosporidium, Plasmodium and Toxopla
sma.

According to the present invention, there is also provided the use of the composition or solution of the present invention for preventing or inhibiting microbial growth.

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

According to the present invention there is further provided a method of preventing, treating or ameliorating a microbial infection, which method 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 invention may be administered topically.

Applicants have found that the compositions according to the invention, in particular the compositions according to the invention in which the unpurified natural substance is honey, have excellent wound healing properties. In particular, chronic wounds lasting from one month to more than a year have begun to visibly heal within a few days of treatment with the compositions of the 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 the compositions of the present invention provide when diluted, eg, by wound exudates. This causes oxygen to be delivered to the wound site.

Oxygen has various important roles in assisting wound healing. The complex wound healing process requires a large amount of energy. Greater energy is required if the wound becomes infected, which means that more oxygen is needed. Oxygen is involved in many mechanisms of the natural healing process. Oxygen is a metabolic support, substrate repair, antiseptic / infection control,
And have a major role in signaling and regulation of cellular responses. Wounds that receive sufficient oxygen generally heal at a greater rate than wounds that do not have sufficient 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 leukocytes are recruited to the wound site, further increasing oxygen consumption.

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 is the matrix formation
As new blood vessels form and the epithelium replaces, it requires the proliferation of various types of cells. Such cellular activity depends on oxygen availability, which allows unhindered breathing. The biosynthetic pathways required to make all biopolymers (eg proteoglycans, structural proteins, etc.) depend on oxygen, which satisfies the basic energy requirements.
A variety of enzymes are required, including matrix metalloproteases, which are also highly consumptive in terms of energy or oxygen demand.

Collagen synthesis is an important part of the above process. Collagen deposition is very important for connective tissue repair and as part of the angiogenic process. Oxygen is a cofactor required for 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 for the completion of healing. This needs to be triggered by the appropriate signal. In a hyperoxic environment, macrophage leukocytes induce this signal, leading to a complex and organized sequence of events including post-tissue degradation collagen formation and organization, endothelial cell migration / colony formation, and angiogenesis. be able to.

Wounding activates the body's natural defenses. Neutrophils gather at the wound site shortly after trauma and release bactericidal reactive oxygen species (ROS) and hydrogen peroxide (H ₂ O ₂ ) to kill bacteria,
Prevent infection. In response to environmental stimuli, macrophages arrive at the wound site, phagocytose foreign bodies, and release vascular endothelial growth factor (VEGF), a highly important angiogenic factor for wound healing. Oxygen has a major role in these events. One of the major microbicidal mechanisms of macrophages and neutrophils is the "respiratory burst," which is the natural activity by which these cells kill microorganisms. Individuals who are defective in the respiratory burst process suffer from bacterial infections. White blood cells require oxygen to carry the respiratory burst effect, and high oxygen concentrations may increase their potency. These leukocytes also have other bactericidal mechanisms that are triggered when the leukocytes swallow microorganisms (phagocytosis). This is an oxygen-dependent process as it entails a considerable amount of energy consumption and thus does not work well in an oxygen deficient environment. Works best in oxygen-rich conditions. Oxygen also kills anaerobes 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 signaling pathways. The molecular signal released by macrophages and triggering angiogenesis is "vascular endothelial growth factor" (VEGF). At high oxygen concentrations, macrophage leukocytes can release VEGF. Genes associated with some other important factors and enzymes are triggered by high oxygen concentrations.

Nitric oxide (NO) production is also recognized as a central signaling and regulates events in wound healing. NO is made by the 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 when rich in arginine and oxygen, allowing NO production at a reasonable 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 and organize themselves to differentiate into structured epithelium.

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

It is believed that in addition to the hydrogen peroxide releasing activity of this composition, some activities of this composition contribute to the wound healing activity of the preferred composition 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 combat pain to minimize scarring. It is considered possible.

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

Have a high osmolality (preferably in the honey-like range, i.e. 0.47-0.
The compositions of the present invention (having an a _w of 7) are believed to promote debridement by the autolytic action of tissue proteases. Due to its strong osmotic effect, the composition of the present invention creates a moist wound environment by draining lymph from wound tissue. This provides a continuous supply of proteases at the interface between the wound bed and the overlying necrotic tissue. This action also cleans the surface of the wound bed from below. Activation of the protease by hydrogen peroxide released by the composition of the present invention in contact with water may also be assisted. The debriding action may also contribute to reducing the bacterial load on the wound by removing necrotic tissue. It is well known that necrotic tissue provides a good medium for bacterial growth and, if left in the wound, increases the risk of infection.

Although the body's inflammatory response is characteristic of the initiation of the healing process, a protracted response can inhibit healing, further damage tissue and make the wound more difficult to manage. Prolonged inflammatory reactions are often accompanied by high levels of exudate. Inhibiting inflammation reduces not only the patient's pain but also the opening of blood vessels, thus reducing edema and exudate. Remove the infection,
The ability of the compositions of the invention to debride is believed to contribute to the anti-inflammatory effect. Antioxidant (
Compositions of the present invention, which preferably include one or more antioxidants present in honey, may also reduce excess inflammation by removing free radicals.

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

The compositions of the invention are also believed to have an immunostimulatory effect mediated by interleukin-1 (IL-1). It is believed that the compositions of the present invention 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 the expression of adhesion factors on endothelial cells, allowing leukocytes to migrate to the site of infection.
It also acts on the body's thermoregulatory center to raise body temperature. Called an endogenous pyrogen. Elevated body temperature helps the body's immune system fight infection. This is the initial 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 defenses against possible infections and by participating in cell and tissue repair and repopulation. It is believed that the antibacterial effect of the composition of the present invention is assisted and complemented by the immunostimulatory effect which aids in the re-growth and repair of damaged tissues and / or cells.

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

According to a preferred aspect 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 management of wound sepsis.

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

Wounds occur when the integrity of any tissue is compromised (eg, skin cracks, muscle tears,
Burns or fractures). Wounds can be caused by an act (trauma) or surgical technique, by an infectious disease,
Or it may be caused by underlying symptoms.

Acute wounds include surgical and traumatic wounds, such as lacerations, abrasions, tears, penetrations or bites, and burns. Acute wounds usually go through an ordered and timely healing process, resulting in sustained repair of anatomical and functional integrity.

Chronic wounds have been present for more than 3 weeks or have not gone through an orderly and timely process resulting in anatomical and functional integrity, failing to produce lasting and functional outcomes, A wound that has not gone through the repair process (Lazarus et al., Arch Der
matol. 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 and make up about 70% to 90% of chronic wounds, often in the elderly. They are believed to be due to venous hypertension caused by improper functioning of valves present in veins that prevent blood reflux. Ischemia results from dysfunction and, combined with reperfusion injury, causes tissue damage leading to the wound. In venous disease, ulcers usually lie in the gaiters area between the ankle and calf, often on the medial aspect of the leg.

Arterial leg ulcers result from diminished arterial blood flow and subsequent tissue perfusion. Atherosclerosis or peripheral vascular disease is the most common cause of arterial leg ulceration.
Leaving the reduced blood supply untreated can result in tissue necrosis in the area supplied by the affected artery. Ulcer growth is often rapid, leading to deep tissue destruction. Arterial leg ulcers can occur anywhere on the lower leg. Often deep and circular with well-defined boundaries.

Diabetic ulcer: Diabetes causes an immune compromise that damages microvessels and impairs adequate oxygenation of tissues, which can result in chronic wounds. Diabetes causes neuropathy, which interferes with pain and cognition of pain. Thus, the patient may initially be unaware of small wounds in the legs and feet, and as a result may not be able to prevent infection or repeated injuries. Pressure also plays a role in the formation of diabetic ulcers. Diabetics are at 15% higher risk of amputation due to chronic ulcers than the general population.

Pressure ulcers usually occur in humans with symptoms such as paralysis, which impairs movement of well-pressed body parts such as the heel, scapula and sacrum. Pressure ulcers are caused by ischemia that occurs when the pressure on the tissue is greater than the pressure on the capillaries, thus limiting blood flow to the area. Muscle tissue requires more oxygen and nutrients than the skin and is most adversely 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 wound scab; reducing bacterial load; ideally immune modification and nutritional improvement It is desirable to use to promote healing. Traditional wound care products cannot achieve all of these effects. Although some conventional treatments have some of these effects, they are quite toxic and are therefore not ideal treatments. However, the composition according to the invention, in particular the composition according to the invention in which the abovementioned substances are honey, possesses all these properties. In addition, the antibacterial activity of the compositions of the present invention can be tailored to be much more efficacious than other honey-based wound care products, and the compositions include antibiotic resistant bacteria such as MRSA, Effective against a wide range of pathogens associated with chronic wounds.

The compositions of the present invention are particularly advantageous in the initial treatment of wounds, especially in patients such as diabetics, where the wounds may be exacerbated. Initial treatment with the compositions of the invention prevents complications that occur in chronic wounds, keeps the patient active, and prevents consumption in coping with complications.

The compositions of the present invention have also been found to be effective in reducing or preventing surgical wound infections.
Surgical wound infections are especially problematic in cesarean section, with a fairly high infection rate of about 10% in cesarean section. The composition of the present invention is easy to apply after surgery. For example, the composition of the present invention may be applied to a dressing, then contacted with the wound and held in place with a second dressing. In Example 40 below, it is demonstrated that a single application of the composition of the invention to post-operative Caesarean wounds resulted in a 60% reduction in surgical site infection rates compared to historical data.

Peripherally inserted central venous catheters (PICC line) 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 threaded intravascularly until the tip enters the vena cava just above the heart. However, infection can occur inside the line or in areas that enter the blood vessels. If an infection occurs, patients are usually given antibiotics. The line may be removed if these do not clear the infection or if the infection is severe. To reduce the chance of infection, a dressing containing an antimicrobial agent such as chlorhexidine or silver may be used at the site of entry of the line.

The compositions of the present invention have been found to be effective in preventing and eliminating colonization of PICC lines by using the compositions of the present invention locally at the site of entry of the line (see below). See Example 39).

The compositions of the present invention can be easily used without any specialized training or complicated equipment. This composition is very suitable for use in the Third World. In the Third World, patients are often highly susceptible to infections, and the materials and equipment used for surgery and wound care are often more 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 dressings, PVP-I or chlorhexidine.

  The compositions of the invention may also be used to treat a transplant site in a donor or recipient.

According to the present invention there is provided a method of treating a wound, which method comprises administering to the wound a composition of the invention.

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

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

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

  According to the invention there is also provided a composition of the invention for the treatment of inflammation.

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

According to the present invention there is also provided a method of stimulating tissue growth, which method comprises administering a composition of the 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 stimulating tissue growth.

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

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

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

According to the invention there is further provided the use of the composition of the invention in the manufacture of a medicament for debriding.

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

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

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

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

The amount of the composition of the invention to be administered will depend on many factors, such as the strength of the composition's antimicrobial properties, and other wound healing properties of the composition, the size of the wound, and the age and condition of the subject being treated. Depends on. However, for many applications 2-100 g or 5-100 g of the composition of the invention
Is considered to be suitable and is preferably 10 to 50 g.

According to a preferred embodiment of the present invention, the composition of the present invention is sterile. The sterile composition is 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 thus the ability to release hydrogen peroxide upon dilution) after sterilization by exposure to gamma irradiation.

Thus, according to the invention there is provided a composition of the invention which has been sterilized by exposure to gamma irradiation.

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

Suitable gamma irradiation levels are 10 to 70 kGy, preferably 25 to 70 kGy, more preferably 35 to 70 kGy.

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

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

Sterile compositions of the present invention stored without exposure to light are expected to remain stable for more than 6 months. For example, such a composition may include high density polyethylene / low density polyethylene (HDP
E / LDPE) tube or polyester-aluminum-polyethylene (PET / Al)
/ PE) May be packaged in a pouch.

The compositions of the present invention are preferably medical grade or medical device grade compositions.
Preferably the unpurified natural material is honey, suitably medical grade or medical device grade honey.

Preferably the composition of the invention comprises creamy honey, more preferably non-pasteurized creamy honey. Such compositions are easy to administer topically, as the large crystals or many large crystals present are minimized by the creaming process.

It will be appreciated that in compositions of the present invention that include honey, it may not be 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 substance. In a particularly preferred embodiment, the composition of the invention comprises
It contains non-pasteurized honey and added purified glucose oxidase.

Accordingly, the present invention provides a storage-stable composition for generating antibacterial activity, the composition comprising non-pasteurized honey and added purified glucose oxidase, the glucose oxidase being sufficient. D-glucose in honey can be converted to release hydrogen peroxide in the presence of various free water, and the composition contains sufficient free water for the glucose oxidase to convert D-glucose. Absent.

The present invention also provides a method for producing a storage-stable composition for generating antibacterial activity, the method comprising contacting unpasteurized honey with purified glucose oxidase. Is capable of converting D-glucose in honey to release hydrogen peroxide in the presence of sufficient free water, and the composition provides sufficient free water for the glucose oxidase to convert D-glucose. Does not include.

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

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

Also according to the present invention there is provided a pharmaceutical composition comprising the composition of the present invention together with a pharmaceutically acceptable carrier, excipient or diluent.

According to certain preferred aspects of the invention, there is provided a dressing comprising the composition of the invention. Suitable dressings include gauze, bandages, tissue paper, films, gels, foams, hydrocolloids, alginates, hydrogels, or polysaccharide pastes, polysaccharide granules or 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 compositions of the present invention may be in the form of solid or semi-solid formulations. Examples of solid or semi-solid formulations include capsules, pellets, gel capsules, powders, hydrogels, pills, pellets or spheres. Alternatively, the composition of the invention may be in the form of a liquid formulation. Examples of liquid formulations include syrups, pastes, sprays, drops, ointments, creams, lotions, oils, liniments or gels. Typical gels include isopropanol gels, alcoholic gels such as ethanol gels or propanol gels, or hydrogels.

The compositions of the invention may be in a form suitable for administration to human or animal subjects. Suitable forms include forms adapted for topical or oral administration. Suitable forms for topical administration include topical ointments, creams, lotions, oils, liniments, liquids, gels or meltable strips. Suitable forms for oral administration include capsules, pellets, gel capsules, pills, pills, pills, troches, dental floss, toothpaste, mouthwash, 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 (eg saliva for oral administration, or by exudates from wounds) to effect peroxidation at the site of administration. Hydrogen may be released.

The compositions of the present invention are 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, humectants, itching relief agents, cleansers, scrubs, detergents, barrier agents, strippers. However, preferably the composition of the invention is free of any antibiotics.

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

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

The compositions of the present invention may be provided in the form of a prophylactic hand barrier solution or hand antiseptic 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, which have advantageous properties for the prophylactic prevention of microbial infections.

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 solves the problem that an antibiotic resistant strain of a microorganism appears.
Overcome by non-toxic means. In this application, the compositions of the invention may be administered topically, eg as a topical ointment, cream, lotion, oil, liniment, liquid and / or gel. The composition of the present invention may be administered as part of a tissue or wipe.

The microbial infection may be an infection of the mouth, eyes, ears, skin, chest or nails. Oral infections may be gum disease, oral ulcers and / or oral hygiene disorders. The oral hygiene disorder may be halitosis and / or gingivitis. Alternatively, the oral infection may be a nasal infection, or a nasal infection, including a nasal Staphylococci infection. Eye infections may include conjunctivitis. The skin infection may be a skin fungal infection. Dermatophyte infections include tinea pedis and / or tinea pedis in humans. In veterinary medicine, dermatomycotic conditions include control of foot-rot, ringworm and zoonotic skin infections. The nail infection may be a fungal nail infection, such as onychomycosis.

The compositions of the present invention may be used to treat skin disorders such as acne, eczema or psoriasis. Acne and eczema may have microbial infectious components that can be treated by this composition, and secondary microbial infections of psoriatic lesions caused by scratches can be treated by the compositions of the invention.

The compositions of the present invention may be used in veterinary medicine. Important veterinary applications include the treatment of microbial infections and the treatment or treatment of wound care or the treatment of burns. Specific symptoms include chronic skin infections in dogs (subcutaneous Staphylococcus infections)
, Otitis externa (ear infection), oral care in animals, Campylobac in chickens
ter infection, scoliosis, enteric microbial infection in pigs, poultry and cattle, Cryptospo
Ridium infections, elimination of common human infections, removal of wound dressings such as horns, and treatment of abscesses. The present invention has the particular advantage of being able to treat microbial infections in veterinary applications without 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 present invention may be provided as a mixture with other compositions or products, or may be provided separately therefrom, for example packaged with other compositions or products as a kit. When the composition of the invention is provided as a mixture with other compositions or products, this other composition or product is
It is preferred that the above enzyme does not contain sufficient free water to convert the substrate of this composition of the invention.

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

According to the present invention, there is provided a kit for preventing, treating or ameliorating microbial infection, or for treating wounds, which kit comprises the composition or solution of the present invention and instructions for administering the composition or solution. Including.

According to the present invention there is further provided a method of preventing or inhibiting microbial growth, which method comprises using the composition or solution of the 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 prevent microbial growth over the shelf life of edible products (such as fruits or vegetables) or other products (bulbs, especially lily bulbs, or expensive plant material including flowers). It may be particularly suitable for prolonging by blocking. If the microbial growth is not prevented or prevented, the product will be damaged. Treatment of edible 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 invention may be used for the prevention or inhibition of Erwinia growth. Erwinia is a genus of the family Enterobacteriaceae and mainly contains phytopathogen species. These species produce enzymes that hydrolyze pectin between plant cells,
Separate the cells. This is a disease called "plant rot". Well-known members of this genus are E. amylovora species, which cause burns in apples, pears and other Rosaceae crops. Erwinia carotovora (Pectobacte
(Rium carotovorum) is another species and has a broad host range including carrots, potatoes, tomatoes, leafy vegetables, squash and other squash species, onions, bell peppers. This species can cause disease in almost any plant tissue in which it invades. It is a very important pathogen as it is a common cause of decay in preserved fruits and vegetables. E. The spoilage caused by carotovora is often referred to as bacterial soft rot (BSR).

Erwinia infection is a particular problem for potato storage. Significant losses can occur when Erwinia occurs. It also causes black rot in the center of the potato. This problem is currently controlled using pharmaceutical products. According to a preferred embodiment of the invention,
The compositions or solutions of the present invention may be used to prevent or prevent potato spoilage 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 invention, the compositions or solutions of the invention may be used to prevent or prevent the growth of insect pathogens, especially Nosema in bees.

Nosema is the most prevalent disease of adult honeybees. It is caused by a unicellular microsporidian parasite that exists in two stages: the long-lived spore stage and the replicative multiplication stage. When adult honeybees ingest spores, the spores grow to the proliferative stage and invade the cells forming the inside of the honeybee intestine. Two species that affect bees, Nosema api
s and Nosema ceranae are described. Nosema apis reduces the lifespan of infected bees. Nosema ceranae transcends species barriers from Asian hosts, and N. It is known to be more pathogenic than apis. Infected foraging bees die away from beehives, thus the continuous loss of bees reduces the transport of food to beehives and subsequently the colony collapses.

Treatment of Nosema with the antibiotic Fumidil B (prepared from Aspergillus fumigatus, the causative agent of Stonebrood disease) inhibits spore replication in the midgut but does not kill the spores. This antibiotic is also very expensive. It should 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 are capable of killing Nosema spores and proliferating Nosema. According to the present invention, there is provided a method of preventing or treating Nosema in honey bees, which method comprises administering to a 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 contact with glucose oxidase in powder form.

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

Example 1
This example describes a preferred process for making 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 Using a heat exchanger, heat the honey for 2 minutes at 80 ° C (if desired, use lower temperature if sufficient to inactivate catalase). Can be, preferably 60
℃ or more). The purpose of this heating process is to pasteurize the honey and to reduce the viscosity of the honey so that it can be filtered to remove any wax particles and honey feathers that may be present in the honey after harvest. , As well as inactivating any catalase in honey that affects the effective generation of hydrogen peroxide.

Next, the pasteurized honey described above is filtered and allowed to stand, and the normal honeycomb temperature (35-40
Allow to cool naturally to (° C). Glucose oxidase is then added at a low concentration (equal to that normally found in honey) to replace glucose oxidase that is naturally present in this honey but is 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 why this mixing cannot be done at room temperature.
The resulting "activated" honey is then stored at room temperature.

There are no modifications to the natural honey composition described above other than the replacement of the inactivated glucose oxidase described above. There is no detectable hydrogen peroxide in the "activated" honey described above.

Dilution of "activated" honey After dilution of "activated" honey, free water becomes available and the glucose oxidase described above initiates the conversion of the glucose present in this honey, so that peroxidation occurs over time. Hydrogen is released. Hydrogen peroxide concentration is less than 2 mmol / liter, but is released for a long time.

Example 2
This example illustrates the results of tests 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.

Cultures were seeded on antimicrobial diffusion agar plates by picking overnight cultures with a cotton swab and placing them on the surface of agar plates. The plate was left at room temperature for 15 minutes. A 7 mm diameter well was drilled on the agar surface. 200 microliters of sample (phenol standard or honey) was placed in each well.

The plates were incubated for 16 hours and the circle of inhibition was measured using a dial caliper (+/- 0.1 mm). The diameter of the zone was recorded, including the diameter of the wells.

Phenol standards were prepared by diluting phenol in purified water to 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 Aspe
rgillus niger origin, supplied by Biozyme UK, activity 240 iu /
mg. Initially 0.5% w / w enzyme was used but this can be reduced to 0.005% w / w enzyme to equal the 20% phenol standard.

Hydrogen Peroxide Detection Merckquant Peroxide Test: Catalog No. 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 for 15 seconds (cat. No. 110011) or 5
After a second (catalog number 110081), the color of the reaction zone is determined using the color region on the label that most closely matches. The corresponding result is read in mg / l H ₂ O ₂ or, if necessary, the intermediate value is estimated.

To confirm that endogenous hydrogen peroxide was not available in the above honey, 50/5
Dissolve in methanol at 0 w / w. Measurement in organic solvents (easily volatile ether):
The reaction zone of the test strip is immersed in the measurement sample (15-30 ° C.) for 1 second. After allowing the solvent to evaporate (3-30 seconds, gently rock the test strip back and forth), soak the reaction zone in distilled water for 1 second to remove excess liquid from the long end of the test strip onto an absorbent paper towel. Or gently blow on the reaction zone 4 times, 3-5 seconds each time. 15 seconds (catalog number 110
011) or 5 seconds (catalog no. 11081), the color of the reaction zone is determined using the color region on the label that most closely matches. The corresponding result is read in mg / l H ₂ O ₂ or, if necessary, the intermediate value is estimated.

Results Figure 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 addition of different amounts of glucose oxidase enzyme formulation). w / w)
, Or samples containing Manuka UMF25 + honey were added to Staphylococcus.
aureus was added to the wells of the seeded agar plate. The results show the diameter of the stop circle recorded for each sample.

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

  The effect of activated honey is bactericidal.

Figure 1b shows the results of the well diffusion assay. In this assay, samples containing pasteurized Ulmo honey with the addition of 0.5% w / w glucose oxidase enzyme preparation were run on MGO Manuka honey, Manuka 25+ honey, and a series of different phenol standards (10
% To 40%) of the sample. The results show the area of the inhibition circle for each sample.

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

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

Example 3
This example illustrates the results of tests demonstrating 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 TINEO honey samples and various phenol standards. Figure 3a shows Staphylo produced by various samples.
2 is a photograph of duplicate agar plates showing the inhibition circle of Coccus aureus. This figure contains a drawing showing the placement of the sample within the wells of the agar plate.
1.10% phenol standard 2.20% phenol standard 3.30% phenol standard 4. Tineo Honey (pasteurized Tineo honey without added glucose oxidase)
5. Tineo inactivation (pasteurized Tineo honey with further heat inactivation)
6. Manuka UMF25 +
7. Tineo20 + (pasteurized Tineo honey with added glucose oxidase (0.005% Biozyme enzyme))

Figure 3a shows Stine honey alone, or Tineo inactivated honey, St
It shows that it did not result in inhibition of aphylococcus aureus growth.
The effect of Tineo honey (Tineo20 +) with added glucose oxidase
Greater than the effect of Manuka UMF25 + and comparable to the 30% phenol standard.

FIG. 3b is a photograph of an agar plate showing the inhibition circles of Staphylococcus aureus produced by various samples. The various samples were arranged as shown below.

1.10% phenol standard 2.20% phenol standard 3.30% phenol standard 4. 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 the Tineo honey activity 25+ and 40+ samples were more effective at inhibiting Staphylococcus aureus growth than the Manuka honey 25+ samples.

Example 4
This example illustrates the results of tests demonstrating the antibacterial effect of activated Ulmo honey and Tineo honey.

Figure 4a shows Staphylococcus au after 24 hours of incubation.
Figure 3 shows the results of three replicates of the well diffusion assay, measuring the circle of growth of reus. The assay was performed as described in Example 2. A sample of phenol in deionized water with a concentration range of 40-10% and two types of manuka (Manuka UMF25 +, Manuka 550MGO)
And a sample of activated Ulmo honey with 0.01% w / w Biozyme glucose oxidase (as in Example 2).

Figure 4b shows the results of a further well diffusion assay. Glucose oxidase (Biozyme formulation described in Example 2) was added to the pasteurized Ulmo honey (0.005%).
Enzyme w / w), stored at 40 ° C. for 24 or 72 hours. The stored sample of activated honey was then subjected to Staphyloco in a well diffusion assay as described in Example 2.
Tested for activity against ccus aureus 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 the activated honey showed the following efficacy in Staphylococcus aureus.
It is shown to be comparable to the 20% phenol standard.

Figure 4c shows the results of a further well diffusion assay in which new and old batches (old batches) of pasteurized Ulmo honey (Biozyme formulation described in Example 2, 0.5% w / w) were used. Received about 4 months before the new batch),
And a Tineo honey that increased the amount of glucose oxidase added (Example 2
Biozyme formulation, 0.0005-0.5% w / w) as described in Manuka 1
Compared to 8+ and 25+ UMF honey, and a series of 10-40% phenol standards.
In Figure 4c, "normal" refers to as-received pasteurized honey, "inactivated".
Refers to pasteurized honey that has undergone further heat inactivation, and "reactivate" refers to pasteurized honey that has added glucose oxidase after further heat inactivation.
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 Figure 4c, the area of the inhibition circle of Staphylococcus aureus is shown. Old batch of pasteurized Ulmo Honey ("Ulmo Old 4 Reactivation")
, New pasteurization batch ("Ulmo new X reactivation" and "Ulmo new Y reactivation"
It is understood that the same level of activity is retained. All these batches showed approximately 2-fold higher activity than Manuka Honey 18+ and higher activity than Manuka Honey 25+. As the amount of glucose oxidase added to the pasteurized Tineo honey was increased, the honey activity increased.

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

Example 5
In this example, the effect of different glucose oxidase enzyme formulations was demonstrated by pasteurizing Tin.
Tested with eo honey. The enzyme formulation used was the Biozyme described in Example 2.
Formulations and ANHUI MINMETALS DEVELOPMENT I / E CO
． , LTD (hereinafter "Anhui" enzyme formulation) from food grade non-GMO
It was the origin glucose oxidase.

FIG. 5a shows the results of a well diffusion assay performed as described in Example 2,
In this assay, pasteurized Tine honey containing no enzyme, or 0.00
Samples of pasteurized Tineo honey containing 5-0.05% w / w Anhui enzyme were compared to samples of various phenol standards (10%, 20%, 30%) and Manuka honey UMF25 +. This result shows that when the amount of enzyme is increased, Staphylococcus au
It has been shown to increase the effectiveness of activated honey against reus. 0.0125%
The w / w enzyme was about the same as the 20% phenol standard and Manuka honey sample,
The 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 this assay, different amounts of enzyme were added to a sample of a commercial batch of pasteurized Tineo honey. The "Tineo" sample is pasteurized honey without the addition of glucose oxidase. The "Tineo inactivated" sample is pasteurized honey that has been further heat treated and contains no added glucose oxidase. The "Tineo20 +" sample contains 0.005% w / w Biozyme glucose oxidase. The results show that the Tineo sample has no activity against Staphylococcus aureus. Tineo20 + sample is Staphylococcus a
It was more effective than manuka 25+ against Ureus and was between the 20% and 30% phenol standards.

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

Glucose oxidase enzyme from Biozyme as described in Example 2 was added to pasteurized Ulmo or Tineo honey at 0.005% w / w and the resulting mixture was 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, Staphylococcus aureu carried out as described in Example 2.
Samples were tested in a well diffusion assay for s. Saved samples 10%, 2
Compared to 0% and 30% phenol standards. The results for the Ulmo honey sample are shown in Figure 6a and the results for the Tineo honey sample are shown in Figure 6b. There was no apparent loss of activity during storage with either type of honey.

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

Powdered honey is from Hon from ADM Specialty Ingredients.
Obtained from i Bake. 0.1% w / w Biozyme glucose oxidase (as described in Example 2) was added to this powdered honey. S carried out as described in Example 2
The antibacterial activity of this mixture was tested in a well diffusion assay against Taphylococcus aureus. The mixture was added to two different wells of the agar plate.
The control was used with honey powder only. The results are shown in Figure 7a. The control showed no activity, whereas 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 treated with ANHUI MINMETALS DEVELOPMENT I / E CO.
． , 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 a clear stop circle.

Example 8
In this example, the antibacterial activity of various different products alone, and the honeys that activated them (
That is, it shows the test results of antibacterial activity when mixed with honey having 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.

The results clearly show that the antibacterial effect is increased when the product is mixed with activated honey.

Example 9
In this example, the antibacterial activity of various different products alone, and the honeys that activated them (
That is, further test results of antibacterial activity when mixed with honey having added glucose oxidase) are shown. 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 the resulting photographs are shown in FIG.

Figure 9a:
1. Famous brand antiseptic cream number 1
2. Famous brand antiseptic cream number 1 + 10% Ulmo 40+
3. Famous brand antiseptic cream number 2
4. Famous brand antiseptic cream number 2 + 10% Ulmo 40+
5. Tineo Honey Activity 20+
6. Manuka Honey 25+
7. Ulmo Honey Activity 20+

Figure 9b:
1. Cough syrup 2. 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+

The results clearly show that the antibacterial effect is increased when the product is mixed with activated honey.

Example 10
This example illustrates a method of sterilizing a composition of the present invention that includes non-pasteurized honey and added purified glucose oxidase.

Target doses of 11.6-14.2 in 10 sealed sachets, each containing 50 g of the above composition.
Gamma irradiation with kGy (dose was 13.1-13.6 kGy as measured by dosimeter) was subsequently tested separately for sterility.

In the sterility test, all work was performed under laminar flow in a clean room. 10g of the above sample
100 ml of sterile tryptone soy broth (TSB: casein pancreatic juice digest, 17 g / L
, Papain digest of soybean meal, 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) and shake Mixed and then transferred to a sterile container. An additional 100 ml TSB was added to remove all sample residue and added to the same vessel. TSB was added to the samples 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.

After the incubation period was completed, one positive result was noted. Demonstration with a sterilization dose of 35 kGy was confirmed.

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

Example 11
This example illustrates the results of tests demonstrating the antibacterial effect of the compositions of the present invention as compared to Manuka UMF25 + Honey and heat inactivated honey (inactive honey). This composition comprises non-pasteurized honey and added purified glucose oxidase (see
Surgery ”), and is sterilized using gamma irradiation.

The well diffusion assay was performed as described in Example 2 using samples of Manuka honey UMF25 +, inactive honey and Surgihoney. FIG. 10 (a) is a photograph of an agar plate showing an inhibition circle of Staphylococcus aureus (ATCC9518) 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. Figure 10
(B) Plate treated with Manuka UMF25 + Honey (top), and Surgih
The plate treated with oney (bottom) is shown.

This result shows that Surghihoney is Staphylococcus aur after sterilization.
It retains significant antibacterial activity against eus and clearly shows that Surgihoney was more effective against Staphylococcus aureus than Manuka honey UMF25 +.

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

The accelerated aging technique is based on the assumption that the chemical reactions involved in material degradation follow the Arrhenius rate function. This function states that increasing or decreasing the temperature by 10 ° C. in a uniform process changes the chemical reaction rate by a factor of about 2 or 1/2. For example, 5.
3 weeks is equivalent to 1 year of storage on the shelf, 10.6 weeks at 55 ° C is equivalent to 2 years, 26
． Five weeks is equivalent to five years.

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

The relationship between real time and accelerated aging is:

Tests, Test Intervals and Required Samples Samples from each batch were tested at the same time intervals. The table below summarizes the tests performed and the total number of samples required at each time point for each batch.

Test method Filled sachet weight: The empty tare has an average tare weight of 1.7 g. Ten sachets were individually weighed on a calibrated laboratory balance and the tare weight was subtracted to record the results.

Pressure Test: Each pouch was tested in a pressure test device according to standard operating procedures. The number of passes and failures was recorded.

Honey pH: 5 bags of ingredients were placed in a glass beaker. H using 3 standard solutions
The anna pH meter was calibrated, then the electrodes were rinsed in deionized water. The pH electrode and electrode probe were dipped in the honey sample and the pH value and temperature were read and recorded from a digital display.

Moisture level: 5 bags of honey were taken at each time point. About 1 ml of sample was taken from each pouch and placed on the sample plate of the refractometer (1 sample at a time). The sample cover was closed and the user observed through the lens while pointing the device towards a light source such as a window. The scale values indicated by the shaded lines were read and the results recorded.

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

Results Filled sachet weight-accelerated aging

Pressure test results-accelerated aging

pH test results-accelerated aging

Moisture content test results-accelerated aging

Color test results-accelerated aging

In Examples 13-35 below, compositions of the invention comprising non-pasteurized honey with added glucose oxidase (in these examples, this composition was designated as "Surgi").
“Honey”) is used to describe the results of treating wounds. Surgiphone
y was provided in closed sachets, each sachet containing 10 g of the composition. The pouch was sterilized using gamma irradiation. The pouch was used from day 0 of treatment. A new Surgihoney was used each time the dressing was replaced. The dressings were changed on a daily basis as noted in the examples, or sometimes more frequently. Surgihoney was either applied to the dressing or applied directly to the wound and then covered by the dressing. Surgih in both cases
The oney was in direct contact with the wound and was covered by the dressing.

Example 13
This example illustrates the results of treating infectious toes using Surgihoney. The results are shown in Fig. 11.

The patient was a 78 year old, diabetic male. The wound on the left foot had developed for a month before the start of treatment and caused a slight amount of pain.
a) Day 0-wound profile: wound: 3x2x1 cm; 99% healthy granulation,
1% green colonization; peeling of surrounding skin; small amount of odorous yellow purulent exudate;
b) Day 5-wound profile: wound improvement; honey 1 sachet each time, second dressing change; metronidazole and amoxicillin; also tea tree oil;
c) Day 10-wound profile: wound improvement; green colony disappeared; wound washed,
The dry skin was removed and an additional honey pouch was used.

Example 14
This example illustrates the results of treating a toe ulcer using Surgihoney.
Results are shown in FIG.

The patient, a 61-year-old diabetic woman, had an infectious toe ulcer on her right foot that had developed mild pain for a month prior to treatment.
a) Day 0-wound profile: wound: 2x2x0.2 cm; 1% tan scab, rest granulation tissue; low amount of yellow exudate;
b) Day 7-wound profile: Wound improvement; Surgihon with daily dressing change.
Use ey0.5 sachet; Flucox;
c) Day 10-wound profile: Wound further improvement; Flucox discontinued.

Example 15
This example illustrates the results of treating a toe ulcer using Surgihoney.
The results are shown in Fig. 13.

The patient, a 61-year-old diabetic woman, had a toe ulcer on both the first and second toes of her 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 exudate;
b) Day 7-wound profile: wound that has healed to the extent that it is not already open; each day, half a honey pouch; Flucox;
c) Day 10-wound profile: wound improvement; presently dry, no scab or exudate; Fl
Stop ucox.

Example 16
This example illustrates the results of treating foot ulcers using Surgihoney.
The results are shown in Fig. 14.

The patient was a 50-year-old diabetic woman with a foot ulcer that developed the month before 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% tantalis lid,
The rest is granulation tissue; very low amount of yellow exudate;
b) Day 7-wound profile: wound improvement; dryness; scab replaced by healthy granulation; reduced size and depth; wound rinsing with saline, honey dressing (0.5 sachet)
Use; no antibiotics;
c) Day 9-wound profile: Greatly improved wound; nearly closed, no exudate present; subsequent use of dressings each with 0.5 sachets.

Example 17
This example illustrates the results of treating foot ulcers using Surgihoney.
The results are shown in Fig. 15.

The patient, 87 years old, was 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 exudate;
b) Day 3-wound profile: wound improvement; honey 1 pouch with dressing change; no pain reported; no antibiotics;
c) Day 6-wound profile: wound improvement; wound bed is now 100% granulation tissue; use additional honey pouches; Diprosalic on dry skin around ulcer.

Example 18
This example describes the results of treatment of diabetic foot ulcer using Surgihoney. The results are shown in Fig. 16.

The patient was male and had a diabetic foot ulcer caused by the irritation of poor quality shoes.
This patient had an ulcer less than 1 month prior to treatment.
a) Day 0-wound profile: red perianal skin and mild pain; 1% scab, 99% granulation; infection and diabetes; low amount of yellow serous exudate; wound size: 2 cm x 2 cm x 0. 2c
m;
b) Day 7-Wound profile: Diagnosis at the hospital is complete, but dressings are changed daily from day 1 to the present. The dressing was changed by the wife of the son, a regular nurse, according to the procedure manual. Wound improvement; 100% healthy granulation; healthy surrounding skin, mild pain; antibiotics used, Flucoxa
cillin, 500 mg, every 6 hours for 7 days; Wound size: 1.5 cm x 1.3 cm
× 0.1.

Example 19
This example illustrates the results of treating foot ulcers using Surgihoney.
Results are shown in FIG.

The patient was a 55 year old male with severe and uncontrolled diabetes. This patient had a deep ulcer at the site of the third toe after amputation.
a) Day 0-wound profile: wound: 3x3x3 cm; 1% tan scab; 1% green colonization; 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 additional sachets; patient discharge; wipes: +++ skin microbiota; no antibiotics.

Example 20
This example describes the results of treating traumatic leg wounds using Surgihoney. The results are shown in Fig. 18.

The patient, a 95 year old female, had a traumatic wound on the lower leg with moderate pain.
a) Day 0-wound profile: wound: 15 x 12 x 1 cm; mostly granulation tissue, but 1% of wound bed is necrotic and black; medium blood exudate;
b) Day 3-wound profile: no change in wound; no more necrosis but approximately the same wound; honey 1 pouch used; wipe sample: Entrecoccus sp. No antibiotics;
c) Day 8-wound profile: wound improvement; less exudate and less necrotic tissue.

Example 21
This example illustrates the results of treating infectious leg wounds using Surgihoney. The results are shown in Fig. 19.

The patient was a 91 year old female with an infectious wound on her lower leg. The surrounding skin is fragile,
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; moderate yellow serous exudate.
b) Day 15-wound profile: wound improvement; size: 4x2 cm; less scab and serous exudation;
c) Day 19-wound profile: wound improvement; using Surgihoney with each dressing.

Example 22
This example illustrates the results of treating foot ulcers using Surgihoney.
The results are shown in Fig. 20.

The patient, a 53 year old diabetic woman, had a chronic diabetic foot ulcer 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 exudate;
b) Day 5-wound profile: wound size improved slightly, wound bed condition remained almost unchanged; unattached granulation tissue was removed; honey pouch with absorbable dressing used; antibiotic No substance;
c) Day 7-wound profile: with wound cleansing, healthy granulation tissue; changing dressing every 2 days using a honey pouch each time;
d) Day 9-wound profile: wound improvement; reduced size, reduced exudate; patient discharge.

Example 23
In this example, treatment results for leg ulcers using Surgihoney are described.
The results are shown in Fig. 21.

The patient was a female, had a leg ulcer that had developed over the year prior to the start of treatment, and had mild pain.
a) Day 0-wound profile: healthy surrounding skin; 1% scab, 99% granulation; medium amount of yellow serous exudate; wound size: 7 cm x 3 cm;
b) Day 7-Wound Profile: Wound Improvement, and Patients Take Use of Community Care; Surgi
I used one sachet of honey; reduced in size, less exudate present; more enduring pain.

Example 24
In this example, treatment results for leg ulcers using Surgihoney are described.
The results are shown in Fig. 22.

The patient, a 77 year old male, was malnourished and dehydrated. anemia. Prostate cancer. This patient had a leg ulcer developing for 6-12 months prior to treatment. Mild pain.

a) Day 0-wound profile: wound: 20 x 10 x 0.5 cm; 1% tan scab,
The rest is granulation tissue, but with moderate green purulent exudate; swab: +++ Col
a mixture of iform and Pseudomonas; antibiotics: Ben Pen and Clin
da;
b) Day 4-wound profile: no wound changes; honey only dressing; Ben Pe
n and Clinda;
c) Day 8-wound profile: wound improvement; all green palates disappeared; no antibiotics; paracetamol.

Example 25
In this example, treatment results for leg ulcers using Surgihoney are described.
The results are shown in Fig. 23.

The patient, a 91 year old female, had a leg ulcer that had developed for 6-12 months.
She was hospitalized and had severe pain.

a) Day 0-wound profile: painful, odorous wounds; infections; scab 10%,
Green colony formation 50%, cellulitis 20%, healthy granulation 20%; Wound: 15 cm x 5 cm x
0.3 cm; exudate: low dose, yellow serum;
b) Day 11-wound profile: mild pain, greatly improved wounds; 100% healthy granulation; no infection; wounds: 10 cm x 5 cm x 0.2 cm; exudate: low dose, bloody red; wound size. 50% reduction.

Example 26
In this example, treatment results for leg ulcers using Surgihoney are described.
The results are shown in Fig. 24.

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

Example 27
In this example, treatment results for leg ulcers using Surgihoney are described.
The results are shown in Fig. 25.

The patient, an 82 year old female, was malnourished and dehydrated. This patient had a leg ulcer developing for 3 months prior to treatment.
a) Day 0-wound profile: wound: 3x3x0.5 cm; wound bed consisting of healthy granulation tissue; small amount of yellow bloody exudate;
b) Day 3-wound profile: wound improvement; wipe specimen revealed ++ skin microbiota; use 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.
Results are shown in FIG.

The patient was an 83-year-old immobile man with low serum albumin. This patient is 3 before treatment
He had a leg ulcer that had developed over a period of months and had moderate pain.
a) Day 0-wound profile: wound: 20 x 18 x 0.5 cm; yellow / brown scab 1%
, 1% green colonization, the rest granulation tissue; medium amount of green serum exudate; slight wound odor;
b) Day 4-wound profile: Wound improvement; using 2 sachets of Surgihoney; ++ Pseudomonas by swab; no antibiotic intake;
c) Day 8-wound profile: wound improved after 2 changes of honey only dressing;
++ Pseudomonas; markedly improved, discontinued honey.

Example 29
In this example, treatment results for leg ulcers using Surgihoney are described.
The results are shown in Fig. 27.

The patient was a 74-year-old man who was a smoker, had type 2 respiratory failure, and was poorly exercised. This patient had septic skin lacerations that had developed over the week prior to treatment and had moderate levels of pain.

a) Day 0-wound profile: wound: 15 × 20 × 0.3 cm; 99% healthy granulation, 1% infected at risk; medium yellow exudate; slight E. coli by swabs Found: Antibiotics: Clindamycin;
b) Day 6-wound profile: wound improved after two dressings changes; honey only dressing; ulcers are now dry; swabs no growth; clindamycin;
c) Day 9-wound profile: wound improvement; cellulitis still remains, but ulcer healed; no re-swab specimens proliferated; clindamycin.

Example 30
In this example, treatment results for leg ulcers using Surgihoney are described.
The results are shown in Fig. 28.

The patient was a 81-year-old male with a heavy and extensive ulcer on the legs and arms. This patient suffered from peripheral vascular disease and was malnourished. The wound had developed over the month prior to treatment. The surrounding skin had peeled off, causing severe pain.
a) Day 0-wound profile: wound: 30 x 28 x 1 cm; 1% yellow / brown scab, rest granulation tissue; large amount of red bloody exudate; +++ E. coli and Pseud by swabs
Omonas mixture found out;
b) Day 6-wound profile: wound improvement; slight Pseudomona; no antibiotics but orormorph for pain;
c) Day 10-wound profile: wound improvement; + Pseudomonas still present;
Severe pain is reduced.

Example 31
In this example, treatment results for leg ulcers using Surgihoney are described.
The results are shown in Fig. 29.

The patient was a 67 year old male with peripheral vascular disease. A leg ulcer had developed for one week before treatment. The surrounding skin was fragile and the pain was moderate.
a) Day 0-wound profile: wound: 1 × 1 × 0.2 cm; yellow / brown scab 1%, rest granulation tissue; small amount of yellow serous exudate at wound;
b) Day 11-wound profile: After a series of dressing replacements where there is no change in the wound, there are now signs of improvement; exudate decreased, wounds smaller; 1 honey pouch at each replacement. Used; Staph aureus ++ found by wiping sample;
c) Day 18-wound profile: no change in wound; 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.
Results are shown in FIG.

The patient was a 43-year-old woman with a large diabetic ulcer present for 1 month. The ulcer was associated with latent osteomyelitis, the bone was debridement and metallized from the calcaneus. The patient had a broken leg and subsequent treatment resulted in a bone infection. The opinion of the primary clinician was that this patient is likely to have a foot amputation.

a) Day 0-wound profile: scab 40%, green colony formation 10%, healthy granulation 50
% Wounds; healthy surrounding skin; moderate pain; moderate amount of yellow purulent exudate; wound 9 cm x 8 cm x
2 cm; no antibiotics;
b) Day 1-wound profile: no change in wound; swabs: E. coli and Pseud
omonas; no antibiotics; no painkillers;
c) Day 14-Wound profile: Wound improvement; 1 sachet of Surgihoney used; Wound slowly granulated and greatly improved; Wipe sample: slight skin microbiota found.

Example 33
This example illustrates the results of treating pressure ulcers using Surgihoney. The results are shown in Fig. 31.

The patient was an 88-year-old diabetic woman who was malnourished. This patient had a pressure ulcer (grade 3) developing for 6 months prior to treatment. You have moderate pain,
The surrounding skin was fragile.
a) Day 0-wound profile: wound size: 0.7 x 0.7 x 0.2 cm; yellow /
1% brown scab, 1% cellulitis tissue, the rest granulation; low amount of yellow pus exudate;
b) Day 3-wound profile: wound healing (wound closed); using one pouch of Surgihoney.

Example 34
This example describes the results of treating infectious axillary wounds using Surgihoney. The results are shown in Fig. 32.

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

Example 35
This example describes the results of treating infections around the entrance of a catheter using Surgihoney. Results are shown in FIG.

The patient was a 41 year old female with cancer. This patient had an infection of the breast area around the entrance of the catheter. The wound was present for less than a week prior to treatment and was mildly painful.
a) Day 0-wound profile: wound: 2 cm x 2 cm; cellulitis 1%, rest granulation tissue; low amount of red serous exudate;
b) Day 6-wound profile: wound significantly improved; no exudate; Surgiphone
Using 1 small pouch of y; wiped specimen but no growth; no antibiotics;
c) Day 14-wound profile: wound improved to the extent that it was no longer a problem.

Example 36
Surgihoney
Surgihoney is a non-pasteurized honey with added glucose oxidase. Three different Surgihoney formulations with different antibacterial activity were made.

S1 Surgihoney: non-pasteurized honey with 0.1% (w / w) added glucose oxidase. The enzyme used was Aspe manufactured by BIO-CAT INC.
It was a food grade glucose oxidase of rgilus niger origin and had an activity of 15,000 units / g. A sealed pouch containing 50 g of S1 Surgihoney was gamma-irradiated with a target dose of 11 / 6-14.2 kGy.

S2 Surgihoney: non-pasteurized honey with 0.1% (w / w) added glucose oxidase. The enzyme used was BBI Enzymes Limi.
Aspergillus niger origin glucose oxidase (GO3B
2) and the activity was 274 units / mg. Unit regulation: 25 degrees Celsius, pH
Amount of enzyme that oxidizes 1 micromolar glucose per minute at 7.0. Contaminants: alpha amylase 0.05% or less, saccharase 0.05% or less, maltase 0.05%
Below, and GO / Cat 2000 or above.

S3 Surgihoney: non-pasteurized honey with 0.25% (w / w) added glucose oxidase. The enzyme used was BBI Enzymes Lim.
It is glucose oxidase (GO3B2) manufactured by ited and has an activity of 274 units / mg.
Met.

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

Example 37
In vitro antibacterial activity of Surgihoney In this example, the disc diffusion method, minimum inhibitory concentration (MIC) and minimum bactericidal concentration (M) were used.
BC) and Surg of a series of wound and ulcer bacterial isolates by time killing measurements
The susceptibility test for ihoney will be described.

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 concentrations that would be reached with topical clinical use. The local concentration of Surgihoney in the wound is
It is estimated to be about 500 grams / L. Staph. Surgih against Aureus
MIC / MBC of oney1 is 31 and 125 g / L, Surgihoney
The MIC / MBC of 3 is 0.12 and 0.24 grams / L.

The sterilization rate depends on efficacy. In the least potent Surgihoney 1, full bactericidal activity occurred within 48 hours against all microorganisms tested. The most effective Sur
For gihoney 3 bactericidal activity occurred within 30 minutes. Maintenance of the Surgihoney inoculum formulation within 1 week showed full bactericidal activity and no bacterial persistence.

Conclusion: Surgihoney has a wide range of potential as a highly active topical treatment that combines the effects of the healing properties of honey with the potent antibacterial activity of bioengineered products for skin lesions, wounds, ulcers and cavities. There is. Highly active against multidrug-resistant bacteria. It is more active than the other honeys tested and is comparable in antibacterial activity to chemical disinfectants.

Superficial wounds and skin ulcers are becoming more common with the increasing age of the population in many countries and with the global epidemics of obesity and type 2 diabetes. In the UK, community nurses spend a lot of time treating leg ulcers, and leg ulcer nurse supervision is essential if they want to maintain standards in their leg ulcer services. . Chronic wounds on the skin are often colonized with bacteria. It is difficult to know when they become pathogenic, and whether they are pathogenic, but bacterial colonization slows tissue healing, even when no overt infection is present. Build a biofilm,
As a result, it may play a role in causing wound scab and malodor.

Tissue viability becomes difficult, especially 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 be sent in an attempt to provide systemic antibiotics. This only contributes to the selection of even more resistant microbes, which is why chronic leg ulcers are so frequent that methicillin-resistant Staphyloco
This is why they are colonized by multidrug-resistant microorganisms such as ccus aureus and Pseudomonas aeruginosa.

Surgihoney has been developed as a prophylactic dressing for wounds. This study examines the in vitro properties of Surgihoney. Although Surgihoney retains all of the established natural honey healing properties, its antibacterial activity can be set at whatever degree of efficacy is required. In this study, Surgihoney1,
The minimum inhibitory concentration (MIC) and the minimum bactericidal concentration (MBC) of 2 and 3 and the time sterilization curve were measured.

Methods Surgihoney provided as potency grades 1, 2 and 3. Surgihon
ey is a semi-solid form and is provided as a sterile pharmaceutical grade in sachets.

Clinical isolates were recovered from soft tissue microbiology samples. Staphylococcus au
Reus isolates 18 strains, methicillin sensitive (MSSA) 12 strains and methicillin resistant (M
RSA) 6 strains, β hemolytic streptococci isolates 6 strains, Lancefield group A
(2), B (2), C (1), G (1), vancomycin resistant E. Faecium-containing Enterococcus spp. 5 isolates of Esch., Including broad spectrum β-lactamase producing strains. coli 6 strain, Klebsiella spp. 2 shares, Serr
atia Marcescens Amp C producing strain 1 strain, Pseudomonas a
eruginosa4 strain, Acinetobacter lwoffii1 strain, Prop
Ionibacterium acnes1 strain, Bacteroides fragil
is1 strain, Candida albicans2 strain, Candida glabra
The ta1 strain and Aspergillus fumigates1 strain were tested against Surgihoney.

Agar diffusion Isosen, which has already been inoculated with the test organism at a concentration that gives semi-confluent growth
6 mm wells were cut in est agar. The test Surgihoney and other honeys from the preliminary study were added to the wells.

First, a preliminary study was carried out to compare Surgihoney potency S1, S2, S3 with various honeys from all over the world, European honey, South American honey, New Zealand honey, Yemani honey, Sudanese honey, and medical honey Medihoney. And an antibacterial coating containing silver (Silver Aquacell) and iodine (Iodoflex). Staphylococcu
s aureus, cut wells in plate and fill with test honey,
Alternatively, in the case of bandages, these were cut into 2 × 2 cm and placed on the surface of the seeded plates.

After preliminary studies, Surgihoney potency S1, S2, S3 were tested alone against a series of bacterial isolates from skin lesions. Wells were filled to the surface with a formulation in which about 2 grams of the three undiluted Surgihoneys of potency were diluted and emulsified with an equal volume of sterile water. After 18-24 hours of aerobic incubation (Candida and Aspe
rgillus spp. Longer then Propionibacterium sp
． And Bacteroides sp. Then anaerobically), the size of the zone was measured.

Minimum inhibitory concentration and minimum bactericidal concentration Surgeony products are liquefied by warming them to 37 ° C and 5 g of them are sterilized with deionized water
Mix with 0 mL. This dilution was considered as the "no dilution" material for serial dilutions. British Society of Antimicrobial Chemotherapy for the practice of Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) (
BSAC) method (Andrews JM. Determination).
of minimum inhibtory conventions. J
Antimicrob 372 Chemother 2001; 48 (Supp 1
): 5-16). Add Surgihoney products to the wells of a microtiter tray,
Serial dilution was performed from no dilution to 1024 times. 75 μL of each honey dilution was added to each well in the strip of the microtiter tray. The undiluted concentration showed a concentration of 250 grams / L and the 2048-fold dilution showed about 0.12 grams / L.

The test organisms were prepared by picking 4 colonies from each culture that were morphologically identical to each organism to produce a 0.5 McFarland density. It was further diluted 1:10.

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

MBCs were measured by subculturing MIC wells and wells near MIC wells in blood agar medium and incubating at 37 ° C. for 18 hours. MBC was the most diluted concentration showing no growth after incubation.

Time kill curve A test organism inoculum 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 (Bijou), a control and 3 test formulations were added to Surgiho
ney1 (S1), Surgihoney3 (S3) or Medihoney (MH)
0.5 g was added. The colony number of the inoculum was determined by making a 1:10 serial dilution and plating on a blood agar plate in 0.1 mL and repeated 3 times.

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

The terminal culture was carried out by inoculating 0.1 ml of the first inoculum into a nutrient broth and surging
Any residual effect of ney was neutralized and incubated at 37 ° C for 72 hours before plating on blood agar to determine test organism survival.

Results Size of inhibition circles In a preliminary comparative study, all Surgihoney potency was compared to M of medical grade honey.
It was demonstrated to have higher antibacterial activity than any other honey tested, including edihoney. The stop circle of S1 was larger than the stop circles formed by all other honeys. The silver coating produced some blocking effect under the coating, but the Su
There was no stopping circle as it existed in rgihoney. Iodine coating material is Staphyl
A large stop circle (about 70 mm) is formed against S. ococcus aureus, and
6 mm) and was equivalent to S3 (67 mm).

In the quantitative inhibition circle size test, full-potency Surgeihoney showed that all bacteria tested,
For both Gram-positive and Gram-negative bacteria, including multiple antibiotic resistant bacteria, as well as fungal species, formed a circle of inhibition in agar diffusion. The size of the stop circle for various types is Surgiphone
y Increased with increasing formulation potency. Table 12. The blocking effect of Surgihoney not only depends on direct contact with the activator as with the silver coating, but also diffuses well across the wells, leading to large blocking circles such as those listed in Table 12. Formed.

MIC and MBC
Surgihoney showed significant antibacterial activity against all isolates tested. MIC
And MBC were well matched among isolates of the same species, regardless of whether the isolate was multidrug resistant or hypersensitive. Table 13 lists the MIC and MBC values for the isolate species tested by dilution ratio, and Table 14 shows the MIC and MBC in grams per liter. The degree of potency increased with Surgihoney grades. The MBC for each isolate was close to the MIC and in most cases was within one dilution.

The local concentration of Surgihoney in the wound is estimated to be about 500 grams / L. Staph. The MIC / MBC of Surgihoney1 against Aureus is 31 and 125 g / L, respectively, and the MIC / MBC of Surgihoney3 is 0.
． 12 and 0.24 g / L.

Time Sterilization Curve Surgihoney quickly kills bacteria. Starting with approximately 105 colony forming units (cfu / mL) per milliliter, the number of cfu / mL in the control was constantly increasing, while in the Surgihoney inoculum, both potions of Surgihon were effective.
After contact with ey, cfu / mL dropped rapidly. By 30 minutes, cfu numbers were often reduced 1000-fold in both S1 and S3 (FIG. 34). In S1, bacterial growth was often undetectable by 2 hours, and in S3 by 30 minutes. Enterococci appeared to be more durable and survived for 48 hours. The bactericidal activity was complete for all organisms as neither organisms were detected in the S1 or S3 inoculum in terminal cultures in nutrient broth followed by plating with blood agar.

Discussion Surgihoney is a natural honey and, unlike many commercial honeys for human consumption, is also organic in the current sense of having no agricultural additives or antimicrobial residues. Surgihoney is not dependent on a specific source of nectar, unlike honey, which relies on a specific source of nectar for high activity, such as manuka. Surgihoney
, The antibacterial activity can be controlled by the compounding process, allowing the production of different grades with a uniform measured efficacy.

This study clearly demonstrated that the efficacy of Surgihoney is active as a very potent antibacterial agent against all species of bacteria and fungi tested. Surgihon
Comparing ey with various honey sources from all over the world, and medical grade honey M
Preliminary studies comparing edihoney showed significantly higher antibacterial efficacy with Surgihoney. Compared to the commonly used topical disinfectants silver and iodine, Surgiho
ney3 produces antibacterial effect as high as iodine coating material, and silver coating material (Aquacel)
It produced a higher antibacterial effect than Ag). The silver coating was only effective at inhibiting bacteria in direct contact with the coating.

In the MIC and MBC tests, Surgihoney not only inhibits the microorganisms, but kills them at concentrations 10-1000 times less than the concentration that would be achieved in topical treatment (estimated to be 500 grams / L). It is shown. Surgiho
The bactericidal activity of ney occurs at concentrations close to its inhibitory activity. Therefore Surgiphone
y can be highly active in multimicrobial inhibition and eradication when used topically on any colonized or superficial wounds or soft tissue cavities. Many chronic wounds are colonized with resistant bacteria and bacterial retention in biofilm formation delays wound healing. Thus, the use of Surgihoney 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 tissues. This is Surgihoney's MIC and MBC
Reflected in the values, these are correspondingly higher than those commonly shown for systemic antibiotics.

The bactericidal activity rate has been shown to be extremely fast according to the time sterilization curve.
It is within 30 minutes for oney3 and within 2 hours for Surgihoney1. This is the case for both Gram-positive and Gram-negative organisms, although Enterococci appear to be slightly more robust. Fungi, Candida spp. , Aspergill
us sp. However, higher concentrations and longer exposures are required to arrest growth and kill organisms.

Surgihony is used as a topical wound dressing on skin lesions and cavities to provide a moist wound healing environment while also reducing microbial colonization to help remove scab and promote granulation and epithelialization 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 analogs also have good antibacterial activity, but have also been reported to be toxic in certain situations. Concerns about the use of chlorhexidine formulations in wound dressings have also increased due to the development and toxicity of antimicrobial resistance.

The clinical utility of Surgihoney appears to be topical use on the skin, in wounds and cavities. Wounds can colonize with bacteria that can biofilm and delay healing. Due to the growing concern about antimicrobial resistance and the lack of new antimicrobial agents, topical agents with broad spectrum antimicrobial activity may play a role in reducing the use of systemic antibiotics in soft tissue lesions. These in vitro studies demonstrated Surgihoney's ability as a wound dressing with high antibacterial activity. Sur
The efficacy of gihoney can be regulated, and Surgihoney also exerts other important functions in wound healing: wet barrier, scab removal, local nutrition, local immune regulation, and cytotoxicity. is not.

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

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

The number of viral (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.

The results show that both the S1 and S2 Surgihoney formulations have potent antiviral activity.

Example 39
Use of Surgihon as a line site dressing Surgihon for prevention of infection of peripherally inserted central venous catheter (PICC line)
To assess the efficacy of ey, S1 Surgihoney was used locally at the line entry site on the arm of 30 patients. About 3 to 8 g of S1 Surgihoney was used in 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 Surgihoney dressings. The results are shown in Table 15 below.

It has been concluded that Surgihoney is an effective antimicrobial agent for use in line site dressings.

Example 40
Use of Surgihoney to Prevent Caesarean Wound Infections Surgical wound infections are particularly problematic in Caesarean sections, with a fairly high infection rate of about 10%. In the UK, 147,726 Caesarean sections each year are associated with Caesarean wound infections (
8-24.6%) and widespread variants across NHS hospitals (13.6-31.9%) are increasing nationwide (Bragg et al., 2010. Variation in ra).
tes of caesarea section among English N
HS trusts after accounting for material
and clinical risk: cross sectional study
． BMJ 2010; 341). Caesarean wound infections are a major cause of long hospital stays, resource consumption, and other morbidity and mortality. Recovery from cesarean section is more difficult in women who develop postoperative wound infections.

To assess the effectiveness of Surgihoney in preventing the infection of Caesarean wounds, S1 Surgihoney was used once once on the wound after surgery. S1 Surgih
About 25 g to 35 g of oney was used in the dressing, which was then contacted with the wound and kept in place with the second dressing. Nearly 200 patients were evaluated over 3 months.

Clinical Evaluation Women who underwent a Caesarean section (CS) between October 2012 and January 2013 were provided with Surgeihoney as a single-use wound dressing when dressing a wound at the end of a surgical procedure. Each patient used 10 g of Surgihoney pouches each. Using aseptic technique, a "non-sterile" surgical assistant emptied the Surgihoney pouch and carefully applied the sterile contents onto the sterile dressing. Then the obstetrician or midwife in the operating room
The dressing was applied to the surgical wound. After the surgical procedure, the midwife in charge completed the evaluation record. The data collected were MRSA status, diabetes history, medication and volume index. Fourteen days after the surgical procedure, the attending midwife also recorded any wound healing problems, especially the presence of exudation, pain and inflammation. In case of inflammation, a wound culture wipe was requested and microbiological results were recorded. S
The Surgical Site Infection (SSI) rate during the 3 month evaluation using the urgihoney coating
It was compared to the infection rate in the 9 months prior to assessment, 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 the percentage of all CS surgical procedures performed.

Results The results are shown in Table 16 below. In the three-month period from October 2012 to January 2013, there were 186 CS cases, of which 102 cases (55%) were urgent. No women were colonized with MRSA. Four (2.23%) had diabetes. 42 (27
． 3%) had a body volume index greater than 25. During the evaluation, 4 out of 186 CSS SSIs were confirmed. It showed an infection rate of 2.15%. An adverse event related to Surgihoney use was reported in one patient, which was a form of wound irritation and resolved in 3 days without further therapeutic intervention. In the last 9 months, 590 CS surgical procedures (23
There are 4 cases depending on selection, and 356 cases are urgent).
The SI was recorded, which showed an infection rate of 5.42%. The reduction 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. Su
The rgihoney dressing was well tolerated with few adverse events reported.

Wound infection rate was reduced by 60.33% when using Surgihoney. Using SSI data from the two groups in this study, the pre-Surgihoney CSS SSI rate (expected) 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 was 8007 cases per year (expected) and 3176 cases per year (observed). ).
The difference, which can potentially be reduced by using Surgihoney, is 4831.
It is a case.

It was concluded that S1 Surgihoney effectively reduces the infection rate of Caesarean wounds after surgery. Prevention of wound colonization with Surgihoney, a drug that is non-toxic to healing tissues and also accelerates the healing process, is a new and potentially important finding, which is a surgical wound. May change the way you deal with. S
Urgihoney provides a clinically effective, cost-effective therapeutic intervention to significantly reduce SSI in women undergoing cesarean delivery.

Discussion This evaluation demonstrated that the highly effective antimicrobial wound dressing Surgihoney can be used as a wound dressing for early CS wounds that prevents infection. As a "natural" product with established wound healing properties, Surgihoney appears to promote wound healing in addition to providing potent antibacterial activity to prevent wound colonization and infection. Some halogen-based chemical disinfectants may provide comparable antibacterial activity, but may delay wound healing (Jan WA. Comparison of
conventional pyodyne dressing with honey
dressing for the treatment of diabetic
foot ulcers. JPMI-Journal of Postgradu
ate Medical Institute 2012; 26 (4): 402-7.
)). Iodine wound dressings are contraindicated in CS (Joint Formulary
Committeee. The British National Formula
ry. London: The Pharmaceutical Press; 20
13), its use is associated with various toxicities (Pietsch and Meakins: Com
applications of povidone-iodine absorption
in topically treated burn patients. 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 .:
Series in a Patient Treated with Conti
Nous Povidone-Iodine Mediastinal Irriga
section. New England Journal of Medicine 19
92; 326 (26): 1784; Colpaert: Iodine toxi.
city as a cause of total atrioventricula
r block in burn patients. Burns 2009; 35
: S45-S6; Ramaswamykanive: Cardiovascula
r collapse following povidone-iodine was
h. Anaesthesia and Intensive Care 2011;
39 (1): 127-30; Lakhal: Povidone iodine:
Features of critical system absorption
． Annales Francaises d'Anesthesie et de
Reanimation 2011; 30 (7-8): e1-8): e1-e3. )
.

Similarly, in Cochrane systemic reviews, whether silver-containing dressings or topical agents promote wound healing and prevent wound infection (Storm-Ve
rsroot et al .: Topical silver for presenting w
found infection. Cochrane Database of Sys
thematic Reviews 2010), or whether they are effective treatments for infectious or contaminated chronic wounds (Vermeulen et al .: Topica
l silver for treating infected wounds (R
view). Cochrane review 2010; (10): 42) has been shown to have insufficient evidence to prove.

Previous systematic reviews have shown evidence of ambiguous benefit for the clinical efficacy of honey as a wound dressing, but this new formulation appears to provide significant clinical benefit in CS patients. (Jull et al .: Honey as
a topical treatment for wounds: The Coch
lane Collaboration, 2009; Jull et al .: Honey a
sa topical treatments for wounds. Cochra
ne database of systematic reviews (Online
e) 2013; 2). In a temporal comparison of wound infection rates, the infection rates were 5.
The above evaluation shows a 60.33% reduction from 42% to 2.15% with the use of Surgihoney.

Healthcare-associated infections are serious and costly medical complications affecting approximately 8% of hospitalized patients.
SI accounted for 14% of these infections and nearly 5% of patients undergoing surgical procedures were found to have developed SSI. SSI is associated with a fairly high morbidity, with more than one-third of post-operative deaths
At least partially related to SSI. Antibacterial prophylaxis is routinely used in many surgical procedures to reduce surgical wound infections. Skin disinfection is also routinely used by surgeons prior to skin incision to reduce skin bacterial load, but antimicrobial dressings are not routinely used. This may be because most topical antiseptics have a deleterious effect on tissue healing.

Surgihoney is a product with strong antibacterial activity, is non-toxic and promotes tissue healing. Topical use of this product on "clean" surgical wounds may actually replace prophylaxis with systemic antibiotics in certain types of surgery. Such developments will help reduce the amount of antibiotics used and the selection pressure of colonizing bacteria.

Caesarean wounds were selected for this evaluation because they were generally healthy, had little or no morbidity, and had an increased CS infection rate. 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 a higher body mass index. Although the use of antibacterial agents in early wound dressings has heretofore not been routine, this evaluation indicates an interesting and effective role for Surgihoney in the prevention of CS wound infections.

Example 41
Use of S1 Surgihoney to Treat Infectious Leg Ulcer The patient was a 52 year old male with diabetes and chronic kidney disease. The ulcer was widespread, painful, odorous, and profuse with significant exudation.

S1 Surgihoney was used as a topical dressing with antibiotics. By day 7, the odor of the wound had diminished, the pain was relieved, and the massive exudation and pseudomonas infection had disappeared. The patient was discharged on day 7.

The photograph of the result is shown in FIG. 36. In FIG. 36, (a) is the first day of treatment;
Day; and (c) shows the 7th day of treatment.

Example 42
Use of S1 Surgihoney to treat pressure ulcer The patient was a 50 year old female patient suffering from spina bifida, a disabled person and unable to move. This patient had a hip to sacral pressure ulcer 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, improvement of the wound was reported. By day 30, the soft tissue cavity had healed almost completely. At this point St
Reptococcus was not detected.

The result 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 Su against Staphylococcus aureus (NCIMB 9518)
The antibacterial activity of rgihoney (SH) and the antibacterial activity of two prototype modified honeys made by Apis mellifera (honey bee) were tested. We also have several modified forms of Sur due to its ability to alter the level of hydrogen peroxide evolution from the sample.
Gihoney was also examined.

Method: Surgihoney (SH) was treated with two denatured honeys, prototype 1 (PT) using a bioassay against Staphylococcus aureus standard strains.
1) and prototype 2 (PT2). Further work considered the rate of hydrogen peroxide evolution from these formulations.

Results: The antimicrobial activity of Surgihoney was shown to be primarily due to hydrogen peroxide evolution. It was shown that the denaturation of Surgihoney resulted in the two more potent honey prototypes producing as much as 2-3 times as much antibacterial activity and up to 10 times as high peroxide activity.

Conclusion: Surgihoney is a clinically available wound disinfectant dressing that exhibits good antibacterial activity. Two additional honey prototypes were shown to have antibacterial activity that could be enhanced due to the demonstrated increased peroxide activity.

Method 1. Measurement of honey activity by bioassay Surgihoney (SH) and two modified honeys, prototype 1 (PT
The antibacterial activity of 1) and prototype 2 (PT2) was confirmed by Staphylococcus au.
Reus (NCIMB 9518) and measured as% phenol equivalent.
Values are calculated from the average values from replicates of 3 samples repeated in 3 days.

Assay method. The agar well diffusion method used was Microbiology Standard.
rd Methods Manual for the New Zealand Da
iry Industry (1982) [Bee Products Standard
rds Council: Proposed standard for measure
ring the nonperoxide activity of honey.
In. New Zealand: Bee Products Standards
Council; 1982. ] From the punch plate assay for inhibitors described in.

Inoculum preparation. Aseptic nutrient broth was used as a blank and diluent, and the culture was adjusted overnight using a cuvette with a 1 cm light path to an absorbance of 0.5 measured at 540 nm.

Assay plate preparation. An assay plate was prepared by inoculating 150 ml of nutrient agar with 100 μl of the culture adjusted to an absorbance of 0.5. The agar was swirled to mix well and poured into a large Petri dish, which was placed on a horizontal surface. Once the agar had set, the plate was inverted upside down and left overnight, 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 before they had a diameter of 7.0 m on the agar surface.
m wells were cut out. 250 μl of test material (sample or standard) was placed in each well.

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

Sample preparation. The original sample solution was prepared by adding 4 g of the sample to 4 ml of distilled water in a universal manner and placed at 37 ° C. for 30 minutes to facilitate mixing. To prepare the second solution, 2 ml of the first sample solution was added to 2 ml of distilled water in a universal manner and mixed for the total activity test, and 2 ml of the first sample solution was added to 2 ml of the catalase solution and the non-peroxide solution was added Mixed for oxide activity.

Preparation of phenol standards. By dissolving phenol in water, standard (w / v) 10%,
30% and 50% phenol was prepared. Phenol standard should be brought to room temperature in the dark before use
Mix well before addition to test wells. Each standard was placed in 3 wells and tested in triplicate. The standard product was stored at 4 ° C. and its expiration date was set to 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 using the samples, the plates were incubated at 37 ° C for approximately 18 hours. The diameter of the stop circle was recorded, which included the diameter of the well (7.0 mm).

Calculation of antibacterial activity of samples. The mean diameter of the well-defined circle around each phenol standard was calculated and squared. A calibration plot of% phenol against the square of the mean diameter of a clear circle was plotted. An optimized line was obtained using linear regression. Using this linear equation, the activity of the honey sample at each dilution was calculated from the square of the average measurement of the diameter of the distinct circle. The figure was multiplied by 4.69 so that dilution was possible (assuming the density of Surgihoney was 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: H ₂ O ₂ is removed by treating the sample with catalase enzyme.

2. Measurement of honey activity by H ₂ O ₂ method Merckquant® 1.10011. And 1.10081 were used to measure activity.

Hydrogen peroxide test kit concentrations were expressed as mg / L H ₂ O ₂ equivalent.

The sample was diluted 1:10 with purified water. After incubation for 5 minutes, all samples were measured for H ₂ O ₂ evolution per hour over a 12 hour period, 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 determined semi-quantitatively by visual comparison of the reaction zone of the test strip to the area of the color scale. The reaction zone of the test strip is dipped in the Surgihoney sample for 1 second and excess liquid is flushed from the strip onto the absorbent paper towel for 15 seconds (catalog number 110011) or 5 seconds (catalog number 110).
081) and then measuring the color produced in the reaction zone, which more closely matches the area of the color scale.

Result 1. Activity evaluation Regarding the antibacterial activity produced by denaturation of the honey sample, the phenol activity at PT1 and PT2 was increased by 2 times and almost 3 times respectively as compared with 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 H ₂ O ₂ Method Denaturation of the prototype is observed to produce hydrogen peroxide activity that is up to 7 and 10 times that of Surgihoney. Results for three samples are shown in FIG.
Shown in. A linear relationship is observed by taking the maximum level of hydrogen peroxide evolution for each of the three honey prototypes and plotting this against total phenol activity (Figure 39).

Discussion The results from this work indicate that the main antibacterial activity of Surgihoney (SH) and 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 could be enhanced, and at 12 hours the values were 7 times and 10 times that of Surgihoney (SH) alone, respectively.
Double. There is a striking linear relationship between the antibacterial activity and the maximum release of hydrogen peroxide from the three honey prototypes.

This peroxide activity provides strong antibacterial activity that is ideally suited for use in wound dressings to treat or prevent wound infections for use in acute or chronic wounds. Although a small amount of catalase was present in the wound and serum levels of catalase in men were reported to be 50 kU / l, the catalase activity in healing wounds was shown to be higher during the first week after wounding. It has been shown that, in practice, the activity level of catalase is restored to its original level two weeks after wounding. Therefore, such a catalase concentration can be used outside the Surgihoney
Or it is very unlikely to affect the antibacterial activity observed with the two modified prototypes, PT1 and PT2.

The ideal properties for an antimicrobial wound dressing 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 the chemical oxidation of cellular components.

Human toxicity of hydrogen peroxide is concentration dependent, and one study argues that specific concentrations for antimicrobial and human toxicity may overlap. In contrast, certain honey formulations provide effective dressing by delivering low concentrations of hydrogen peroxide to the wound continuously over time, rather than supplying large amounts of hydrogen peroxide at one time. It is an antibacterial agent and has been shown not to have such toxicity. Indeed, there is convincing evidence that the application of physiological concentrations of hydrogen peroxide to mammalian cells stimulates biological responses and activates specific biochemical pathways in these cells.

Apparently Surgihoney and two modified prototypes, PT1 and PT2
Is an antimicrobial coating that provides effective hydrogen peroxide release for over 24 hours.

Conclusion Surgihoney and two modified prototypes, PT1 and PT2,
It has been shown to have potent antibacterial activity against phylococcus aureus standard strains. These antibacterial activities have been shown to be due to hydrogen peroxide. This activity is variable in size and can be explained by hydrogen peroxide activity. These modified honeys provide effective, non-toxic, and easy to administer coatings.

Example 44
This example presents a preferred honey-based composition of the present invention. This composition comprises honey and added glucose oxidase (referred to as "active honey").

Alcoholic gel Carbopol 940 0.3%
Triethanolamine 0.4% (necessary for pH adjustment and stability adjustment)
Active honey (5+) 65%
Ethanol 25.0%
Sufficient water Usage: For 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
This example presents a preferred honey-based composition of the present invention. This composition comprises honey and added glucose oxidase (referred to as "active honey").

Aqueous Spray 25 ml plastic bottle contains:
Active honey 10g
Triton CF 0.1g
Maltodextrin or corn starch 1g
Usage: general wounds and burns

Triton CF acts as a surfactant in this composition. Maltodextrin or corn starch act as a film forming agent.

Example 46
This example presents a preferred honey-based composition of the present invention. This composition comprises honey and added glucose oxidase (referred to as "active honey").

Non-aqueous spray 70% active honey
Propylene glycol 30%
Usage: wound in general

Example 47
This example presents a preferred honey-based composition of the present invention. This composition comprises honey and added glucose oxidase (referred to as "active honey").

Coating-Plastic pouch Various calcium alginate coatings 10 cm x 10 cm, 20 cm x 20 cm
Active honey 10% w / w
Usage: Debridement, relatively exudative wounds. This composition provides a relatively moist environment.

Example 48
This example presents a preferred honey-based composition of the present invention. This composition comprises honey and added glucose oxidase (referred to as "active honey").

Honey and painkillers Active honey 99%
Ibuprofen 1%
Usage: Deal with wounds at all stages and provide pain management in wound healing

Example 49
This example presents a preferred honey-based composition of the present invention. This composition comprises 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. The other ingredients are
It can be varied, for example to obtain different levels of moisture absorption.
Usage: Wrap in foil to prevent water absorption. Provides PVP-granulation. CMC-Provides moisture drying power.

Example 50
This example presents a preferred honey-based composition of the present invention. This composition comprises honey and added glucose oxidase (referred to as "active honey").

Debridement Contains active honey (eg, 5-50% active honey), optionally, preferably 1-2%
Reconstituted aqueous solution also containing an analgesic, preferably ibuprofen, w / w.
Usage: For example, it is placed in a dispenser (pump type dispenser, etc.) and used as a flushing liquid for wounds after debridement to provide early initiation of wound healing and pain relief.

Example 51
This example presents a preferred honey-based composition of the present invention. This composition comprises honey and added glucose oxidase (referred to as "active honey").

Foam coating Polyurethane backing for absorbing exudate Island of calcium alginate soaked in active honey Silicon to prevent adhesion to wounds Usage: A foam-based dressing to aid in absorption of wound exudate.

Example 52
This example presents a preferred honey-based composition of the present invention. This composition comprises honey and added glucose oxidase (referred to as "active honey").

Treatment of onychomycosis A plaster containing active honey (5+) in the cavity of the foam Hydroxypropyl cellulose glycerol isopropyl alcohol Citric acid monohydrate Usage: For the treatment of fungal nail infections

Example 53
This example presents a preferred honey-based composition of the present invention. This composition comprises honey and added glucose oxidase (referred to as "active honey").

Throat spray Honey and glycerol, preferably 5-20% honey Usage: Throat sore. The formulation can be placed in a dual spray dispenser or reconstituted with water before use and then sprayed.

Example 54
This example presents a preferred honey-based composition of the present invention. This composition comprises honey and added glucose oxidase (referred to as "active honey").
Pomade activated honey 25%
White petrolatum Light liquid paraffin Talc Kaolin Zinc oxide Use: For the treatment of infectious eczema.

Example 55
This example presents a preferred honey-based composition of the present invention. This composition comprises honey and added glucose oxidase (referred to as "active honey").

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

Example 56
This example presents a preferred honey-based composition of the present invention. This composition comprises honey and added glucose oxidase (referred to as "active honey").

Cream honey 15%
Carbomer 2.63%
Dimethicone 0.13%
Disodium lauryl sulfosuccinate 0.05%
Disodium edetate 0.13%
Glycerol 5.26%
Colloidal silica 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 in the drawings of the present application.