EP3142687A1

EP3142687A1 - Antimicrobial composition comprising a carbohydrate, glucose oxydase and zinc oxide

Info

Publication number: EP3142687A1
Application number: EP15736533.9A
Authority: EP
Inventors: Carmen MALEPEYRE; Anaïs RAYNAUD; Fabien QUERO
Original assignee: Melipharm
Current assignee: Melipharm
Priority date: 2014-05-14
Filing date: 2015-05-13
Publication date: 2017-03-22
Also published as: FR3020948A1; EP4032542A1; WO2015173522A1; US20170072024A1; FR3020948B1

Abstract

The invention relates to a composition comprising a mixture consisting of at least one carbohydrate, glucose oxydase and zinc oxide. Said topically applied antimicrobial composition is especially useful for the care and treatment of skin and mucosal disorders.

Description

ANTIMICROBIAL COMPOSITION

The present invention relates to a composition comprising a mixture of at least one carbohydrate, glucose oxidase and zinc oxide as an antimicrobial composition, and its use on the skin or mucous membranes.

It is very often necessary to use antimicrobial compositions for topical application, in particular for the treatment of wounds, or for the treatment of cutaneous microbial infections or pathologies of the skin with a microbial origin.

Among the products used for the treatment of wounds, creams, ointments or dressings with silver, iodine or dressings containing honey are particularly known.

Silver dressings are often very costly and generally exhibit cytotoxicity to dermal and epidermal cells (Lansdown ABG, A Pharmacological and Toxicological Profile of Antimicrobial Agents in Medical Devices, Adv Pharm Sci 2010, 2010). : Epub 2010 Aug 24, Burd A, Kwok CH, Hung SC, et al A comparative study of the cytotoxicity of silver-based dressings in cell monolayer, tissue explant, and animal models.Wound Repair Regen 2007; ): 94-104) and used in pediatrics, they can cause elevations of the blood level of money.

Iodine dressings are also cytotoxic and their antimicrobial activity decreases sharply in the presence of organic matter (pus, fibrin or necrosis). In addition, there are risks of sensitization with contact dermatitis and allergy to these iodine-based products. For several years, honey has become a product of interest in the medical field for the treatment of certain infections in wounds, however there are large variations in their antimicrobial properties according to their origins. However, honeys with significant antimicrobial activity often have cytotoxicity to skin cells. It is therefore very difficult to find a honey that is both highly antimicrobial and non-cytotoxic. The applications WO-2005/112852, WO-2007/045931, US2004 / 121020 or WO-2004/000339, for example, describe honey-like compositions, dressings or derivatives of dressings based on manuka honey associated with texturizing agents or other active ingredients. The compositions described make it possible to decontaminate the wounds and thus promote their cicatrization.

However, these compositions are not necessarily satisfactory in terms of effectiveness, in particular on migration and cell proliferation, necessary for healing. In addition, the manufacture of these compositions is often complex and expensive, in particular because of the use of complex manufacturing processes (association with interfaces) and / or the addition of active ingredients to specific and expensive honeys. (like for example manuka honey).

FR2995532 describes honey-based compositions which overcome these disadvantages and which in addition to their antibacterial effectiveness are capable of inhibiting the formation of biofilms, of having a bactericidal effect vis-à-vis biofilms already formed, and of prevent the adhesion of bacteria.

The objective of the present invention is to provide a different antimicrobial composition at least as effective as those described in FR29995532.

For this purpose, the invention provides a composition comprising a mixture consisting of at least one carbohydrate, glucose oxidase and zinc oxide, especially for use as a topically applied antimicrobial product.

The mixture according to the invention has a significant and surprising synergistic effect with regard to the existing compositions, which makes it possible to propose a composition having an antimicrobial activity which exceeds that of currently existing honey-based products, without variation of cytotoxicity.

Other features and advantages will become apparent from the detailed description of the invention which follows.

The invention therefore relates to a composition comprising a mixture consisting of at least one carbohydrate, glucose oxidase and zinc oxide.

The carbohydrate is very preferably at least glucose. Preferably it is glucose alone or a mixture of carbohydrates including glucose.

The source of carbohydrate may be especially honey. The honey may be natural honey or artificial honey or a mixture of several natural and / or artificial honeys. By artificial honey is meant at least one sugar. It can be a combination of at least two sugars.

In the case where the carbohydrate source is natural honey it may be preferentially honey thyme and / or honeydew and / or buckwheat and / or manuka and / or mixtures thereof. It may be for example a honey as described in the application FR1258722.

In the case where the carbohydrate source is artificial honey, it is preferably composed mainly of glucose and / or fructose and / or sucrose.

According to a particularly preferred embodiment, the carbohydrate is glucose.

The carbohydrate (s) preferably represent 1% to 99% by weight of the total weight of the composition, more preferably between 1 and 60%.

The composition according to the invention additionally comprises one or more carbohydrates, glucose oxidase and zinc oxide.

Glucose oxidase is an enzyme that catalyzes the oxidation reaction of glucose to hydrogen peroxide in the presence of water. This enzyme can be of natural or biotechnological origin. It is mainly present in microbial sources and obtained by fermentation of fungi such as Aspergillus niger.

The glucose oxidase is present at a dose of at least 0.025 mU (mili-unit) per gram of composition according to the invention, preferably between 0.5 m U and 50 U per gram of composition.

If the carbohydrate is honey, which can produce hydrogen peroxide thanks to the glucose oxidase it contains naturally, the amount of glucose oxidase that it contains will not be sufficient to obtain the activity and the efficiency related to the presence of glucose oxidase as defined in the invention. Supplementation of honey and other constituents with glucose oxidase at a dose of at least 0.025 mU is required to achieve the results described in the invention. In the case where the composition contains honey, the composition is well supplemented with glucose oxidase, in addition to that which may optionally be contained in honey, and this additional dose is preferably at least 0.025 mU. Zinc oxide is a mineral compound of formula ZnO used in many applications, especially in topical preparations. For the present invention, the size of the zinc oxide particles used is very preferably less than 50 μm.

The zinc oxide preferably represents between 0.1% and 40% by weight of the total weight of the composition, even more preferentially between 0.1% and 10%. The composition according to the invention may be composed exclusively of carbohydrate (s), glucose oxidase and zinc oxide or contain other constituents.

It may in particular also comprise, in addition to the mixture, one or more constituents chosen from vanillin, caprylyl glycol, pentylene glycol, 1,2-hexanediol, a propolis extract, methylglyoxal, glycerol, propylene glycol, polyethylene glycol, pectin, gums, carrageenan, xanthan, alginate salts, cellulose and its derivatives, starch and its derivatives, chitosan, triglycerides, vitamins, butters, waxes, glyceryl stearate, petrolatum, paraffins, kaolin, bentonite, lanolin.

It may also contain excipients chosen from excipients which are dermatologically compatible and / or applicable to the skin and the mucous membranes in order to obtain a composition in the form of a powder, or in liquid form such as a lotion, or semi-solid such as a cutaneous powder, a spray, a cream, an ointment, a gel, a paste, an emulsion, a suppository, an egg. It may be, for example, excipients such as sugars and sugar derivatives, polysaccharides (pectins, starch and derivatives, alginates and derivatives, chitosan and derivatives, cellulose derivatives, gums, beta-glucans), synthetic polymers, waxes, natural or non-natural oils, butters, mineral products (silica, talc, clays, titanium oxide), glycerides and other fatty esters, surfactants, water, ethanol propylene glycol, butylene glycol, glycerol, sorbitol, extracellular matrix compounds such as hyaluronic acid or collagen, hydrocarbons and silicones, proteins and peptides and other excipients known to those skilled in the art .

The composition may also contain auxiliary formulation additives such as surfactants, gelling agents, absorbing agents, humectants, solvents, leveling agents, stabilizers, complexing agents, rheological modifiers, preservatives, antioxidants and antimicrobials, dyes and fragrances.

The composition according to the invention may be sterile, that is to say that it has undergone a sterilization process. Sterilization is preferably carried out by gamma radiation.

The composition according to the invention can be obtained by simple mixing of the constituents. Preferably, it is obtained by a homogeneous mixture of powder or by dispersion of the powders in a honey composition. The composition according to the invention may be in various powder or liquid or semi-liquid forms, especially cutaneous powder, spray, lotion, cream, emulsion, gel, ointment, egg, suppository.

The composition is used as an antimicrobial product for topical application, in particular as a health product or cosmetic composition. Indeed, the mixture according to the invention acts in synergy for both:

to limit the microbial proliferation: the composition is capable of destroying germs on the skin, in particular at the level of wounds,

inhibit the formation of biofilms on the skin and / or destroy biofilms already formed on the skin, in particular at the level of wounds,

prevent or fight against the adhesion of microorganisms on the skin, especially at the level of wounds.

Decreasing the microbial load indirectly promotes healing. In addition, the composition according to the invention may also make it possible to promote the proliferation of fibroblasts for a healing effect.

The effects obtained with the mixture of the three constituents of the composition according to the invention are much better than those obtained with each of the constituents alone or mixtures of each of the constituents two by two. There is a surprising synergistic effect.

At the microbial level, each of the three constituents of the composition according to the invention can make it possible to very slightly limit microbial proliferation, but when they are used together, the decrease in microbial proliferation is greater than the sum of the limitations induced by the microbial growth. constituents alone and there is a surprising synergistic effect. According to the invention, the composition can therefore be advantageously used as a topically applied health product on the skin or mucous membranes in humans or animals, very preferably as:

- health product, in particular a medical device for therapeutic purposes intended to be applied to irritation or skin lesions, in particular for:

• the treatment of first and second degree burns,

The treatment and / or prevention of post-operative scar disunions,

• the treatment and / or the post-operative prevention of the residual cavities of the pilonidal sinuses, • treatment of infected surgical scars after flattening,

• the treatment of ulcers and pressure ulcers during the budding phase,

• treatment of dermabrasion, traumatic and / or surgical wounds,

· The treatment of acute and chronic wounds,

• the treatment of oncological wounds,

• the treatment of ostomy sites,

• the treatment of superficial wounds.

composition intended for the treatment of irritations and / or infections of microbial origin, in particular scratch wounds, diaper rashes, acne, dermatitis, impetigo, boils, folliculitis, paronychia and mycoses, especially as a cosmetic composition,

- oral gel, in particular for treating canker sores and mucositis,

- stick or lip balm for treating chapped skin,

- cream for the hands and feet intended in particular for the treatment of crevices and frostbite,

- balm for the prevention, care and / or treatment of nipple crevices,

- vaginal gel to treat skin lesions and microbial infections,

- suppository or rectal cream to treat rectal lesions and irritations.

By medical device for therapeutic purposes is meant a health product intended to be used in humans or animals for purposes including prevention, control, treatment and / or mitigation of a disease, an injury, condition or skin irritation.

If it is a health product for the healing of cutaneous lesions in the form of gel, cream or ointment, the composition according to the invention may in particular be applied according to the following use protocol:

- rinse the wound and dry gently,

covering the entire wound or the secondary dressing with a film of the composition according to the invention,

- cover compresses and an occlusive dressing during the first applications for very exuding wounds. In the debridement phase, and depending on the condition of the wound, it is advisable to repeat the dressing once or twice every 24 hours.

In budding phase, it is advisable to redo the dressing every 48 to 72 hours.

In epithelialization phase, it is advisable to repeat the dressing every 3 or 4 days.

The invention is now illustrated by composition examples and test results demonstrating its effectiveness.

EXAMPLES OF COMPOSITION ACCORDING TO THE INVENTION

Example 1

The composition of Example 1 is a powder consisting of:

• 98.9% Glucose

• 1% ZnO

• 0.1% Glucose oxidase

The composition is obtained by mixing the compounds.

Example 2

The composition of Example 2 is an ointment consisting of:

• qsp 100% blend of natural honeys

• 25% triglycerides

• 11% white beeswax

· 7,5% of shea butter

• 1.83% tocopherol acetate

• 5% glyceryl mono stearate

• 5% ZnO

• 0.01% Glucose oxidase

the percentages being percentages by weight relative to the total weight of the dry matter of the composition.

The composition is obtained by the implementation of the following steps:

finely dispersing the powders in the honey,

to heat the mixture to less than 40 ° C,

- heating the lipophilic phase (wax, butter and glycerides) to about 70 ° C to melt the solids, when the lipophilic phase has reached the temperature of 40 ° C, add the tocopherol acetate and the mixture of honeys and powders,

mix the ointment until cool.

Example 3

The composition of Example 3 is a paste consisting of:

• qs 100% artificial honey

• 2% ZnO

• 0.1% Glucose oxidase

The com position is obtained by mixtures of the compounds.

Example 4

The composition of Example 4 is a paste consisting of:

• qsp 100% buckwheat honey

• 1% ZnO

• 0.1% glucose oxidase

The composition is obtained by mixing the compounds at room temperature.

Example 5

The composition of Example 5 is a gel consisting of:

• qs 100% honeydew honey

• Glycerol 15%

· PEG 1500 17.5%

• Pectin 4%

• ZnO 10%

• Glucose oxidase 1.5%

The position is obtained by:

- mixing of the powders, then at 40 ° C addition of glycerol and honey,

dissolution of the PEG at 70 ° C,

cooling at 40 ° C,

adding the honey base in PEG at 40 ° C, mix until cool. EVALUATION OF THE EFFICACY OF THE COMPOSITION ACCORDING TO THE INVENTION

Attempts have been made to demonstrate the antimicrobial efficacy of several compositions according to the invention on the one hand and compare this efficacy to that of the constituents alone.

1. In vitro antibacterial and anti-biofilm efficacy

For the performance tests, several formulas have been tested:

- formula 1: Honey alone

- formula 2: Honey + 0.1% glucose oxidase

- formula 3: ZnO alone 1%

- formula 4: Honey + ZnO 1%

formula 5: honey + 0.1% glucose oxidase + 1% ZnO (not irradiated)

formula 6: honey + 0.1% glucose oxidase + 1% ZnO + excipients (formula in normal aging = 4 months, irradiated)

- Formula 7: Honey + 0.1% glucose oxidase + 1% ZnO + excipients (accelerated aging formula 92 days = 1 year, irradiated)

formula 8: glucose oxidase 0.1%

- Formula 9: glucose alone

Formula 10: glucose + glucose oxidase 0.1%

formula 11: glucose + ZnO 1%

Formula 12: glucose + glucose oxidase 0.1% + ZnO 1%

The microbial strains used for the tests are the bacteria:

P. aeruginosa

- S. aureus

The tests are carried out on microplates for evaluation of the MIC (Minimum Inhibitory Concentration).

at. Bacterial proliferation tests

The bacterial proliferation in the presence of the products to be tested is determined by a 96-well microplate method. Each well is inoculated with 50 μl of test bacterial suspension made in Muller-Hinton broth (MH) + 150 μl of diluted test product solution. The bacterial concentration in the well is set at 10 ⁶ CFU / mL. The positive control (50 μl of bacterial suspension + 150 μl of MH medium) corresponds to 100% bacterial proliferation. The negative control (200 μl of culture medium) corresponds to 0% bacterial proliferation.

Each sample is tested in triplicate. The optical densities (OD) are read a first time at time 0 (T0) at 450 nm. The plates are then incubated 24h at 37 ° C. with stirring. At the end of the incubation the OD is re-measured at 450 nm (T24). The percentage of proliferation is calculated as follows:

Proliferation = (OD _T 24 - OD _T o) / (OD _T 24 Control + - OD T0 Control +) The results obtained are presented in Tables 1 and 2 below:

Table 1: Influence of the assets used and formulas developed from these assets on the proliferation of P. aeruginosa. The formulas are tested in microplaaues at concentrations of 1.5% 3%, 6% and 9% (v / v). The proliferation percentages are calculated from the positive control corresponding to the untreated medium (culture medium alone = 100% proliferation).

Formulas tested 1.5% 3% 6% 9%

Formula 1 92% 98% 48% 6%

Formula 2 76% 86% 40% -7%

Formula 3 119% 125% 99% 55%

Formula 4 107% 107% 39% -4%

Formula 5 (Invention) 72% 30% -4% -11%

Formula 6 (Invention) 64% 44% -31% -30%

Formula 7 (Invention) 86% 41% -14% -15%

Formula 8 96% 96% 91% 86%

Formula 9 100% 206% 206% 113%

Formula 10 96% 98% 89% 79%

Formula 11 156% 86% 60% 41%

Formula 12 (Invention) 94% 63% 31% 0%

It is found that formulas 5, 6, 7 and 12 cause a sharp decrease in the proliferation of P. aeruginosa. This decrease is much greater for all formulas containing at least one carbohydrate, GOD and ZnO compared to formulas containing either at least one carbohydrate and GOD (formulas 2 and 10) or at least one carbohydrate and ZnO (formulas 4 and 4). 11). For formula 12, bacterial growth decreases by 79% and 41% respectively at the concentration of 9% v / v to reach CM I. CM I goes from 9% v / v to 6% v / v for formulas where the carbohydrate source is honey. Table 2: Influence of actives used and formulas developed from these assets on the proliferation of S. aureus. The formulas are tested in microparticles at concentrations of 1.5% 3%, 6% and 9% (v / v). The proliferation percentages are calculated from the positive control corresponding to the untreated medium (culture medium alone = 100% proliferation)

Formulas tested 1.5% 11 il he:

Formula 1 96% 49% -4% -29%

Formula 2 75% 37% 7% -34%

Formula 3 109% 117% 105% 75%

Formula 4 90% 17% -13% -44%

Formula 5 (Invention) -2% 4% 3% -30%

Formula 6 (Invention) 4% -17% -10% -58%

Formula 7 (Invention) 7% -4% -37% -57%

Formula 8 81% 72% 54% 46%

Formula 9 96% 86% 83% 72%

Formula 10 92% 95% 72% 65%

Formula 11 78% 25% 8% -8%

Formula 12 (Invention) 52% 2% -23% -53%

It is found that formulas 5, 6, 7 and 12 cause a sharp decrease in the proliferation of S. aureus. This decrease is much greater for all formulas containing at least one carbohydrate, GOD and ZnO compared to formulas containing either at least one carbohydrate and GOD (formulas 2 and 10) or at least one carbohydrate and ZnO (formulas 4 and 4). 11). For formula 12, bacterial growth decreases by 95% and 25% respectively at the concentration of 3% v / v to reach CM I. CM I goes from 9% v / v to 6% v / v for formulas where the carbohydrate source is honey. b. Antibiotic activity

The antibiofilm activity in the presence of the products to be tested is determined by a 96-well microplate method. Each well is inoculated with 50 μl of test bacterial suspension made in Tripticase soy broth + 1% glucose (TS + 1% Glu) + 150 μl of diluted test product solution. The bacterial concentration in the well is set at 10 ⁶ CFU / mL. The positive control (50 μl of bacterial suspension + 150 μl of TS + 1% Glu medium) corresponds to 100% bacterial proliferation. The negative control (200 μl of culture medium) corresponds to 0% bacterial proliferation. Each sample is tested in triplicate. The plates are then incubated 24h at 37 ° C. At the end of the incubation the supernatant is removed, the microplate is washed 3 times with 200 μί / ρυίΐ5 of distilled water. The plate is then incubated at 60 ° C. for 30 minutes. A 0.5% crystal violet (CV) solution is added and the plate is incubated for 20 minutes at room temperature. At the end of the incubation the solution of CV is removed and the microplate is washed again 3 times with 20C ^ L / well of distilled water. The plate is then dried overnight at 60 ° C and finally 10 μL / well of a 33% acetic acid solution is added to the wells. The plate is then photographed. The wells in which the biofilm has formed are purple (+) whereas the wells in which no biofilms have formed are clear (-).

The results obtained are shown in Tables 3 and 4 below:

Table 3: Influence of the assets used and formulas developed from these assets on the proliferation of P aeruainosa. The formulas are tested in microplaaues at concentrations of 1.5% 3%, 6% and 9% (v / v). The biofilms are revealed by a 0.5% crystal violet solution. The positive control corresponds to the culture medium control + P. aeruainosa, the negative control corresponds to the culture medium without bacteria

Formulas tested 1.5% 3% 6% 9%

Formula 1 + + +

Formula 2 + + + -

Formula 3 + + + +

Formula 4 + + + -

Formula 5 (invention) + + - -

Formula 6 (invention) + + - -

Formula 7 (invention) + + - -

A decrease in anti-biofilm activity at 6% v / v was observed with the association {honey + glucose oxidase + ZnO} on P. aeruginosa.

Table 4: Influence of the active ingredients used and the formulas developed from these actives on the proliferation of S. aureus. The formulas are tested in microplaaues at concentrations of 1.5% 3%, 6% and 9% (v / v). The biofilms are revealed by a 0.5% crystal violet solution. The positive control corresponds to the culture medium control + S. aureus, the negative control corresponds to the culture medium without bacteria

Formulas tested 1.5% 3% 6% 9%

Formula 1 + +

Formula 2 + + - -

Formula 3 + + + +

Formula 4 + + - -

Formula 5 (invention) - - - -

Formula 6 (invention) - - - -

Formula 7 (invention) - - - -

A decrease in antibiofilm activity of 1.5% is observed with the association {honey + glucose oxidase + ZnO} on S. aureus. vs. Demonstration of the synergistic effect

PRINCIPLE OF THE BERENBAUM TEST

The Berenbaum test makes it possible to determine additive, synergistic or antagonistic effects between two products mixed with cell contacts (Berenbaum, 1977). This test was developed to determine, using experimental data inserted in a mathematical formula, the impact of several mixed compounds. It is applicable to many areas of biology. This test is applied here to {miel + GOD} and {ZnO} from the bacterial proliferation results obtained on strains S aureus and P aeruginosa.

The analysis of proliferation results is based on a search for equivalent activity for each compound. For example, the activity sought is proliferation of 50%. The equivalent active dose required to obtain 50% proliferation (in the case of our example) is designated by Aeq for {honey + GOD}. Béq is the equivalent dose needed to achieve the same effect (50% proliferation) with {ZnO}.

In a second step, the mixture of the two compounds makes it possible to determine the doses of each product to be introduced together to obtain the same effect (50% proliferation) as with Aéq or Béq. The required dose of {honey + GOD} as the dose of A and that of {ZnO} as the dose of B.

Subsequently, it is possible to introduce these data into the following equation:

A B

Berenbaum + Index

eq eq

If the result of this equation is 1, the effect of the 2 compounds will be additive

If the result is less than 1, we will be able to synergize.

On the other hand, if the result is greater than 1, the compounds are designated as antagonists.

RESULTS OF THE BERENBAUM TEST Table 5: Doses of active ingredients necessary to obtain a bacterial proliferation of 50%

The Berenbaum index is less than 1 for both bacterial strains. The formula according to the invention therefore shows a synergistic action of the active compounds.

2. Comparison of antibacterial efficacy with honey before peroxide activity

Tests were carried out with honey having an equivalent peroxide activity (peroxide activity of 0.025 μg / g of honey per hour) to which was added zinc oxide, with or without glucose oxidase.

The operating procedure is the same as in 1 (a).

The results obtained on the proliferation of Pseudomonas aeruginosa in the presence of various honey compositions diluted to a concentration of 6% are presented in Table 5bis below:

Table 5bis: Importance of the presence of GOD to effect bacterial overgrowth

GOD ZnO% bacterial growth o u / g 0% 126% o u / g 0.5% 85% o u / g 1% 58% o u / g 2% 62%

0.5 U / g 1% 0%

5 U / g 1% 0%

0.5 U / g 2% 0%

5 U / g 2% 0% It is found that the sole peroxide activity of the honey is not sufficient to obtain the antibacterial activity and that the presence of glucose oxidase is necessary to obtain a synergistic effect.

3. Cytotoxicity Tests In order to evaluate the cytotoxicity of the formulas, a cell count test in the presence of honey and a composition according to the invention (formula 6) was carried out on fibroblasts. Protocol

The products are diluted to a concentration of 5.12% v / v of the sample.

The procedure is as follows:

Primary cultures of human fibroblasts PAF 08052 are seeded in 48-well microplates at a rate of 10,000 cells per well with a volume of 500 μί complete culture medium. The cells will then adhere to the bottom of the wells for 24 hours. The medium will then be replaced by the test product diluted in serum-deprived culture medium. The product is tested at a rate of 500 μl per well and incubated for 48 hours at 37 ° C. + 5% CO2. The cells are then peeled off with trypsin and then counted on Malassez's slide with an exclusion dye (Trypan blue). The percentage of fibroblastic proliferation is determined relative to a control not treated with the composition.

Results

The results are shown in Table 6.

Table 6: Cell enumeration on fibroblasts in the presence of honey-based formulas. The middle control corresponds to the culture medium alone used to dilute the samples

(dilution in% v / v). The cells were treated for 48 hours

The composition according to the invention (formula 5) does not exhibit significant cytotoxicity.

4. In vivo efficacy

The in vivo efficacy of the composition according to the invention was evaluated on a panel of patients treated in care facility (retirement home, hospitals). Formula 5 {Honey + 0.1% GOD + 1% ZnO + Excipients} was tested in 6 patients with infected, necrotic (n = 6) and / or necrotic (n = 4), debridement wounds ( n = 1) or not (n = 5).

The results of the study are presented in Table 7 below:

Table 7

We aknowledge :

• 83% effectiveness on fibrin

• Effective on infected wounds

• 100% effectiveness on necrosis

• 100% effectiveness on inflammation

• 100% efficiency in accelerating the healing rate

• 100% of the tested panel considers that the formula tested is more effective for detersion compared to honey alone.

Claims

A composition comprising a mixture of at least one carbohydrate, glucose oxidase and zinc oxide, the glucose oxidase being present in an amount of at least 0.025mU per gram of composition.

2. Composition according to claim 1 characterized in that the mixture consisting of at least one carbohydrate, glucose oxidase and zinc oxide contains between 1 and 99% by weight of carbohydrate relative to the total weight of the composition.

3. Composition according to one of the preceding claims characterized in that the mixture consisting of at least one carbohydrate, glucose oxidase and zinc oxide contains between 0.5 mU and 50 U glucose oxidase per gram of composition.

4. Composition according to one of the preceding claims, characterized in that the mixture consisting of at least one carbohydrate, glucose oxidase and zinc oxide contains between 0.1 and 40% zinc oxide by weight by weight. relative to the total weight of the composition.

5. Composition according to one of the preceding claims, characterized in that the at least one carbohydrate is glucose.

6. Composition according to one of claims 1 to 4, characterized in that the mixture consists of honey, glucose oxidase and zinc oxide.

7. Composition according to claim 6, characterized in that the honey is honey of natural origin.

8. Composition according to claim 6 or 7, characterized in that the honey is thyme honey and / or honeydew and / or buckwheat and / or manuka and / or mixtures thereof.

9. Composition according to claim 6, characterized in that the honey is artificial honey or a mixture of natural honey (s) and artificial (s).

10. Composition according to claim 9, characterized in that the artificial honey consists mainly of glucose and / or fructose and / or sucrose.

11. Composition according to one of the preceding claims, characterized in that it also comprises at least one constituent selected from vanillin, caprylyl glycol, pentylene glycol, 1,2-hexanediol, a propolis extract, petrolatum paraffins, lanolin, pectins, starch or starch derivatives, alginates or alginates derivatives, chitosan or chitosan derivatives, cellulose or cellulose derivatives, gums, carrageenans , xanthans, beta-glucans,, methylglyoxal, synthetic polymers, waxes, natural or non-natural oils, butters, mineral products, glycerides and other fatty esters, surfactants, water, ethanol, propylene glycol, polyethylene glycols, butylene glycol, glycerol, sorbitol, extracellular matrix compounds such as hyaluronic acid or collagen, hydrocarbons and silicones, proteins and peptides.

12. Composition according to one of the preceding claims, characterized in that it is in the form of a powder, in liquid form or in semi-liquid form.

13. Composition according to one of the preceding claims, characterized in that it is in the form of powder for cutaneous use, spray, gel, ointment, cream, emulsion, suppository or egg.

14. Composition according to one of the preceding claims, characterized in that it has been treated by gamma irradiation.

15. Composition according to one of the preceding claims, for its use as a topically applied antimicrobial product.

16. Composition according to one of claims 1 to 14 for its use as a topically applied health product.

17. Composition according to one of claims 1 to 14 for its use as a topically applied anti-inflammatory health product.

18. Composition according to one of claims 1 to 14, for its use as a health product for the treatment of skin irritations or lesions.

19. Composition according to one of claims 1 to 14, for its use as a health product for the treatment of burns, post-operative scar disunions, residual cavities pilonidal sinuses, infected surgical scars after flattening, ulcers and eschars, dermabrasion, traumatic and / or surgical wounds, chronic wounds, acute wounds, oncological wounds, stoma sites or superficial wounds.

20. Composition according to one of claims 1 to 14, for its use as a health product for the treatment of scratching lesions, diaper rashes, acne, dermatitis, boils, folliculitis, paronychia mycoses or impetigo.

21. Composition according to one of claims 1 to 14, for its use as a vaginal gel for treating skin lesions and microbial infections, as a rectal cream or suppository for treating rectal lesions and irritations, as hand cream and feet for cracks and frostbite, balm for prevention and / or treatment of nipple cracks, as an oral gel to treat canker sores and mucositis, or as a stick or lip balm for treating chapped skin.