COMPOSITION FOR THE TREATMENT OF SKIN AND MUCOUS MEMBRANE DISEASES

Abstract

The present invention relates to a composition comprising as active ingredients an extract of Cetraria islandica, silver and/or a salt thereof and hyaluronic acid and/or a salt thereof. The present invention further relates to the use of such composition for the treatment of skin and/or mucous membrane diseases.

Claims

1. An association of at least an extract of Cetraria islandica, silver and/or a salt thereof and hyaluronic acid and/or a salt thereof.

2. A composition comprising the association of claim 1 and one or more excipients.

3. The composition according to claim 2, wherein the extract of Cetraria islandica is present in an amount between 0.01% and 30% w/w.

4. The composition according to claim 2, wherein silver and/or a salt thereof is present in an amount between 0.0001% and 20% w/w.

5. The composition according to claim 2, wherein the hyaluronic acid and/or a salt thereof is present in an amount between 0.005% and 30% w/w.

6. The composition according to claim 2 for topical use, in particular for nasal, vaginal, anal, auricular, cutaneous or buccal administration.

7. The composition according to claim 2 in in a form selected from cream, powder, solution, suspension, ointment, gel, and spray.

8. A method of treating skin and/or mucous membrane diseases comprising administering the composition of claim 2 to a subject in need thereof.

9. The method of claim 8, wherein the composition is administered to oral, nasal, anal, auricular, genital and/or vaginal mucous membranes.

10. A method of treating microbial infections of the skin and/or mucous membranes comprising administering the composition of claim 2 to a subject in need thereof.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0040] As above mentioned, the present invention relates to an association comprising as main active principles at least an extract of Cetraria islandica, silver and/or a salt thereof and hyaluronic acid and/or a salt thereof.

[0041] The main active ingredients of the association of the invention are described hereinafter.

[0042] Silver

[0043] The academic medicine has known and has used for some time silver, under the form of salts or nanoparticles (AgNps), which had and still have a therapeutic utility thereof, mainly linked to the organism defence from infections and diseases mediated by bacteria, viruses and mycosis. The effectiveness of the silver compounds is based upon the capability of biologically active Ag+ ion to damage irreversibly the key system of the enzymes in the membrane of the pathogens. In the herein described compositions and kits, Ag+ ion or Salts thereof, colloidal silver, nanoparticle silver, citrated silver, complexed silver could then be used.

[0044] Extract of Cetraria Islandica

[0045] Under the wording “at least an extract of Cetraria islandica” in the context of the present description it is meant that the composition can include an extract of lichen belonging to the species Cetraria islandica or Lichene islandico. The extract could be prepared according to the procedures known in the state of art, for example, it could be a dry extract, or prepared by means of extraction in water or in alcohol solution, and optionally lyophilized. Preferably an extract of Cetraria islandica, with ratio extract/drug 1 to 4 obtained from the thallus will be used.

[0046] The person skilled in the art could adapt the amount of extract used in the preparation of the formulations to be administered depending upon needs. The doctor could be able to identify the optimum dosage for the subject to be treated based upon age, sex, weight and general health status. Then, the dosage of the single active principles can be adapted, even during the period of ingesting the association or composition of the invention depending upon the results obtained in time.

[0047] Hyaluronic Acid

[0048] The hyaluronic acid (HA) also known for its re-epithelizing action is one of the active ingredients of the composition of the present invention, wherein even the Salts thereof, such for example sodium hyaluronate, could be used. The hyaluronic acid could be for example in form of gel or solution, with low or high molecular weight.

[0049] The term hyaluronic acid with high molecular weight in the present document describes a hyaluronic acid having a molecular comprised between 1.0 million Daltons at about 4.0 MDa. For example, the high molecular weight of hyaluronic acid could have a molecular weight of about 1.0 MDa. In another example, it can have a molecular weight of about 2.8 MDa. The term hyaluronic acid with low molecular weight in the present document describes a hyaluronic acid having a lower molecular weight at about 1.0 MDa. Hyaluronic acid with low molecular weight can have a molecular weight comprised between about 200,000 Da (0.2 MDA) to less of about 1.0 MDa, for example, between about 300,000 Da (0.3 M Da) to about 750,000 Da (0.75 MDA). The conjugated hyaluronic acid for example could have a viscosity between 50,000 and 300,000 mPa.

[0050] The inventors have detected the optimal pro-dose dosages (or per single dosage unit) of each substance of the association to obtain synergic therapeutic effects.

[0051] In particular there are the best effects when the extract of Cetraria islandica is present in an amount between 0.01% and 30%, preferably between 0.05% and 15%, still more preferably between 0.1% and 10%; the silver and/or a salt thereof is present in an amount between 0.0001% and 20%, preferably between 0.0005% and 10%, still more preferably between 0.0006% and 5% and hyaluronic acid and/or a salt thereof is present in an amount between 0.005% and 30%, preferably between 0.01% and 20%, still more preferably between 0.05% and 10% with respect to the total weight of the final pharmaceutical form including all excipients.

[0052] The present invention further relates to compositions comprising the association according to any one of the herein described embodiments and one or more suitable excipients.

[0053] The composition can include other components, such as for example excipients, carriers, stabilizers, preservatives and the like and/or other active principles.

[0054] The compositions according to any one of the embodiments provided in the present description can be formulated in any form and by any administration route and associated to any other component, in a variety of ways.

[0055] According to a preferred embodiment, the compositions of the invention are compositions for topical use, for example for nasal, buccal, vaginal, genital, anal, auricular, cutaneous application.

[0056] The compositions will be for example in solid, liquid or gel form, for example, cream, powder, solution, suspension, ointment, gel, spray.

[0057] Suitable excipients can be selected among those usually known in the state of art and include, but they are not limited thereto: diluents, lubricants (for example magnesium stearate, stearic acid, waxes), dispersants, surfactants (for example sodium lauryl sulphate and polysorbates), flavouring agents, adsorbents (for example silica gel, talcum, starch, bentonite, kaolin), glidants and anti-adherent agents (for example talcum, colloidal silica, corn starch, silicon dioxide), dyes (for example iron oxides), opacifiers (for example titanium oxide), antioxidants, binders (for example rubbers, starch, gelatine, cellulose derivatives, sucrose, sodium alginate), disaggregating agents (starch, microcrystalline cellulose, alginic acid, crospovidone), plasticizers (for example ethylcellulose and other cellulose derivatives, acrylates and methacrylates, glycerol and sorbitol), preservatives (for example parabens, sulphur dioxide), viscosifying agents, emulsifiers, humectants, wetting agents, chelating agents and mixtures thereof.

[0058] The compositions of the present invention, for example, will be a medical device, a cosmetic, a dietary supplement, a nutraceutical, dietetic or nutritional composition, a food product, a beverage, food, a nutraceutical, medicated food, food for special medical purposes or a pharmaceutical composition.

[0059] The compositions according to any one of the herein described embodiments could be used both by the human beings and by animals.

[0060] The combination of the above-mentioned active principles could be used formulated in one single composition according to the various above-described embodiments or in a kit including the different separate ingredients, for example, in single compositions formulated in suitable oral dosage form as defined above for the sequential or contemporary administration of the different active principles.

[0061] Therefore, the present invention further relates a kit of portions which include the different active principles of the association according to any one of the herein described embodiments separated and formulated in suitable form of oral dosage for the sequential or contemporary administration of the different active principles.

[0062] The action of the single components and of the association set forth by the present invention was evaluated by means of in vitro and/or in vivo tests which, besides, highlighted that the composition of the invention is particularly effective thanks to the (synergic) potentiation action of its components.

[0063] Therefore, the present invention further relates to the association, to the compositions and to the kit, according to any one of the herein described embodiments, for use in the treatment of microbial infections, for example for infections of S. Aureus, L. Monocytogenes; Salmonella T., Shigella S., E. Coli and P. Aeruginosa, Candida. In particular in the treatment of the irritation and/or disease of the mucous membranes, caused by microbial infections of bacterial or fungus nature, for example associated to pathological states and to a weakening of the immune defenses. The association of the above-mentioned active principles could be formulated in one single composition or in a kit according to the various above-described embodiments and prepared for example by mixing the active principles selected with possible other excipients as it is known to the person skilled in the art.

EXAMPLES

[0064] Hereinafter some not limitative examples of the compositions according to the present invention are reported. In the examples the percentages, if present, are to be meant as percentages by weight. Modifications or variations of the herein exemplified embodiments, obvious to a person skilled in the art, are comprised in the enclosed claims. Where it is not specified, the remaining part of the composition are excipients

Example 1

[0065]

TABLE-US-00001 Active principle % in composition Cetraria islandica 0.5 Silver 0.06 Hyaluronic acid sodium salt 0.1

[0066] Pharmaceutical Form: Spray for Nasal Application

Example 2

[0067]

TABLE-US-00002 Active principle % in composition Cetraria islandica 0.1 Silver 0.005 Hyaluronic acid sodium salt 0.2

[0068] Pharmaceutical Form: Spray for Buccal Application

Example 3

[0069]

TABLE-US-00003 Active principle % in composition Cetraria islandica 0.5 Silver 0.06 Hyaluronic acid sodium salt 0.05

[0070] Pharmaceutical Form: Spray for Ear Application

Example 3

[0071]

TABLE-US-00004 Active principle % in composition Cetraria islandica 0.1 Silver 0.0006 Hyaluronic acid sodium salt 0.01

[0072] Pharmaceutical Form: Spray for Cutaneous Application

Example 4

[0073]

TABLE-US-00005 Active principle % in composition Cetraria islandica 0.5 Silver 0.06 Hyaluronic acid sodium salt 0.1

[0074] Pharmaceutical Form: Cream for Nasal Application

Example 5

[0075]

TABLE-US-00006 Active principle % in composition Cetraria islandica 0.05 Silver 0.0006 Hyaluronic acid sodium salt 0.01

[0076] Pharmaceutical Form: Cream for Vaginal Application

[0077] Experimental Evidence

[0078] The effectiveness of the association of Cetraria islandica, silver and/or a salt thereof and hyaluronic acid was evaluated according to known experimental models. In particular, for evaluating the different activities of the composition of the three active ingredients, in vitro and/or in vivo assays, known to the scientific literature, can be used. In order to demonstrate the antimicrobial effectiveness of the association, of particular interest for the prevention and/or the treatment of bacterial pathologies and surface mycosis, for example, in vitro assays on plate according to the method by spreading (Kirby-Bauer test, or methods of the small disks) or by micro-dilution result to be suitable, but even by means of MBEC Assay designated to quantify the biofilm. The bacterial strains in exponential growth phase selected for the study could be for example: S. Aureus and L. Monocytogenes such as Gram+; Salmonella T., Shigella S., E. Coli and P. Aeruginosa such as Gram−; at last, as fungus strain Candida A or others considered relevant for the purpose of the composition set forth by the present invention.

[0079] The synergic activity of the composition set forth by the present invention can be evaluated even through evaluation of MIC and MBC on bacterial or fungus strains.

[0080] The synergic effect obtained by using the three active ingredients in association can be highlighted by testing the association for its antimicrobial effect on male mice (n=60; 20 to 30 g) thereto, after anaesthesia, wounds with full cutaneous thickness on the back were performed and therethrough a certain bacterium (for example S. Aureus) was inoculated. The rats were divided into groups, including control group. On established days (for example 0, 7, 14 and 21 of the experiment) photos were taken. Based upon the group, the control group, each active ingredient singularly and then even the mixture of the three ingredients of interest for the present composition were applied on the wound on day 0, to evaluate the way in which the association of silver, hyaluronic acid and lichene islandico speeds-up significantly the healing process of infected wounds. On the contrary, to demonstrate the anti-inflammatory effectiveness of the composition, of particular interest for the prevention and/or treatment of symptoms associated to phlogistic processes, in vitro assays result to be suitable which evaluate the capability of inhibiting the release of pro-inflammatory cvtokines such as for example 1L-1, IL-5 and TNF-α and histamine and of inducing the one of anti-inflammatory cytokines such as IL-10, as well as of inhibiting the expression of enzymes such as 5-lipoxygenase and COX-2/PGE2 induced by IL-1 in primary cultures of human cells (for example mononuclear cells of the peripheral blood, such as monocytes and lymphocytes and on fibroblasts or other types of cells) and on pig leukocytes or other type of cell known to the person skilled in the art by means of ELISA test, Western Blotting or flow cytometry. By means of an in vivo model, instead, it was possible to test the association for its anti-inflammatory effect on BALB/C mice (female), to the ear thereof allergic dermatitis by contact was induced by means of inoculation of dinitrofluorobenzene (DNF). The topical treatment was performed once a day for 4 days and provided (for example for groups of about 15-30 mice): the control, each active ingredient singularly and then even the mixture of the three ingredients of interest for the present composition. The swelling and erythema to the ear were evaluated day by day. After 4 days of treatment, the mice were sacrificed and the ear of each one of them was removed for the histological and immunohistochemical examinations, TUNEL Assay and extraction of total RNA for RT-PCR examination. In the immunohistochemical examination the expression of cytokines was evaluated, the extraction of RNA was used to test the levels of the genes of interest involved in the various inflammatory reactions. At last, in order to demonstrate the antioxidant effectiveness of the composition, of particular interest for the prevention and/or treatment of symptoms associated to situations of stress in the organism, after physical damages, in vitro assays result to be suitable such as biochemical analyses on serum and spectrophotometric techniques, which evaluate the capability of incrementing the activity of GPX (Glutathione peroxidase) and of CAT (catalase), the active enzymes against the oxidative damage of the molecules inside the organism.