ES2702398A1 - Compositions for wound healing (Machine-translation by Google Translate, not legally binding) - Google Patents

Abstract

Compositions of asiaticoside. The present invention relates to aqueous compositions comprising asiaticoside, hyaluronic acid and sodium hyaluronate, as well as their use in promoting the healing of wounds, in particular wounds of the oral cavity, and in the treatment and/or prevention of inflammations. . (Machine-translation by Google Translate, not legally binding)

Description

DESCRIPTION

Compositions for wound healing

Field of the invention

The present invention relates to aqueous compositions comprising asiaticoside, hyaluronic acid and sodium hyaluronate and to their use in the healing of wounds, in particular wounds of the oral cavity, and in the treatment and / or prevention of inflammations.

BACKGROUND OF THE INVENTION

The oral cavity is delimited by the upper part, by the palate; on the lower part by the tongue and the floor of the mouth; laterally by the cheeks, by its anterior side by the lips and later, it is opened to the pharynx by the so-called isthmus of the fauces, this being delimited by the uvula, the palatoglossal arches and the posterior area of the tongue. In the interior of the oral cavity are the alveolar arches, two, superior and inferior, where the dental pieces are implanted, 32 in a complete mouth of an adult. These arcs define two spaces within the buccal cavity: the vestibular area: between cheeks, lips and alveolar arches and the mouth itself, delimited by the alveolar arches, tongue and palate.

The set of the oral cavity is covered with mucosa, called buccal mucosa, which isolates the structures, internal tissues of the organism from the external environment. The buccal mucosa is structured in three different parts, from the outside to the inside:

1. Epithelium: It is the most superficial layer and is formed by a coating epithelium, which is formed by flat cells that are organized forming a multilayered tissue in general without cornifying, except on the back of the tongue, the hard palate and the gums .

2. Conjunctiva: It is the intermediate layer on which the epithelium rests and contains the structures that give it nutrition and innervation and others that are specific to the function of the area where it is located.

3. Submucosa: This is the deepest layer of the oral mucosa.

To maintain the integrity of the oral mucosa, it must remain hydrated. Among the various functions of saliva produced by the salivary glands that are located in depth to the mucosa, is the hydration and maintenance of the structural integrity of the buccal mucosa.

The buccal mucosa, covering the oral cavity, takes different names and functions depending on its location: lingual mucosa, palatal mucosa, periodontal mucosa, and so on.

The mucous membranes, in general, are more friable tissues than the cutaneous layer, skin, which covers the human body from the outside. The cavity and buccal mucosa are exposed throughout the day to multiple stimuli, related to the functions it performs or of another nature. Part of these stimuli attack the buccal mucosa, being able to give rise to injuries that concern one or more layers of the buccal mucosa: traumatisms, hygienic maneuvers, medical manipulations, or others that follow with a solution of continuity of the mucosa and exposure of the internal environment To the exterior.

The natural course of these injuries that occur with tissue continuity solution, is the repair and restoration of the affected organ function, through a healing process avoiding inflammation.

The success of healing is determined by the presence of a favorable environment that allows the body to restore the pre-conditions to the injured tissue. The wounds in the oral cavity represent a problem for the quality of life of the subject who suffers, making it difficult to perform usual actions, such as chewing or phonation necessary in daily life.

The healing process comprises three phases: inflammatory, fibroblastic and remodeling. In each of the phases, different cellular cells and mediators intervene, in order to repair and isolate the internal environment from external aggressions. After the fibroblastic phase, it has been possible to isolate and preserve the external environment of the intern. In the therapeutic field, substances are known and used that modulate the first two phases of healing, in order to accelerate the restoration of the integrity of the organism.

Asiaticósido is a triterpenoid present in Centella asiatica . Extracts of this plant have been used extensively in the repair of the skin. The main active compounds of Centella asiatica are Asiaticósido, Asian acid, Madecasósido and acid madecásico The chemical structures of asiaticósido (I) and madecasósido (II) are shown below.

(I) (II)

The asiaticoside, in several published studies, shows that it increases the production of hydroxyproline, the constituent amino acid of collagen, by 56%, as well as increasing the tensile strength, the collagen content and the reepithelialization in the regenerated tissue of wound models. [Shukla, A. et al., Journal of Ethnopharmacology, 1999, 65 (1), 1-11]. This tissue regeneration allows to reduce the continuity solution more than 50% after a week of use of topically applied asiaticoside. The use of asiaticoside has also been described in promoting wound healing of both hard tissue and soft tissue of the oral cavity [Nowwarote, Phyrotherapy Research, 2013, 27, 457-462; Weinreb et al., Journal of Clinical Periodontology, 2015, 42, 288-293].

Hyaluronic acid is a linear polysaccharide of the unsulfated glycosaminoglycan type with a molecular weight of 4,000 to 20,000,000 Da. The structure of hyaluronic acid consists of disaccharide units of glucuronic acid and N-acetylglucosamine connected by alternating P1-3 and | 31-4 bonds. Hyaluronic acid, is a linear polysaccharide and is naturally present in the body as part of the extracellular tissue matrix connective, synovial fluid, embryonic mesenchyme, vitreous humor, skin, mucous membranes and in many other organs and tissues of the body. Hyaluronic acid is an important element in soft periodontal tissues, such as the gum or buccal mucosa and the periodontal ligament, and in periodontal hard tissues, such as alveolar bone and cementum. Given the wide distribution in the oral cavity of hyaluronate, it is proved that this active has a great involvement in the resolution of cicatricial processes, in all stages of healing (inflammation, regeneration or fibroblastic and remodeling) [Bertolami, CN, & Messadi, DV, Critical Reviews in Oral Biology & Medicine, 1994, 5 (3), 311-337].

Furthermore, it is known to use hyaluronic acid to promote the healing of buccal tissue, as well as to alleviate the inflammatory processes that can occur both in infectious processes and in surgery of the oral cavity [Casale et al., International Journal of Immunopathology and Pharmacology, 2016, 29, 572-582].

However, there is a need to develop new compositions that promote wound healing, in particular of the oral cavity, which have high efficacy, permanence of the active ingredients in the oral cavity and stability.

Summary of the invention

The inventors have found that the combination of asiaticoside, hyaluronic acid and sodium hyaluronate has surprising efficacy in contributing to the healing of wounds, in particular wounds of the oral cavity, such as those caused by surgical intervention, and in the treatment and / or the prevention of inflammations. In addition, the compositions of the present invention exhibit unexpected physical and / or chemical stability to light and / or temperature and / or pH changes

Therefore, in a first aspect, the present invention relates to an aqueous composition comprising asiaticoside, hyaluronic acid and sodium hyaluronate.

In a second aspect, the present invention relates to a composition according to the first aspect for use in medicine.

In a third aspect, the present invention relates to a composition according to the first aspect for use in promoting wound healing, in particular of the oral cavity.

In a fourth aspect, the present invention relates to a composition according to the first aspect for use in the treatment and / or prevention of inflammation, in particular inflammation of a tissue of the oral cavity, such as gums.

Detailed description of the invention

Compositions of the invention

In a first aspect, the present invention relates to an aqueous composition comprising asiaticoside, hyaluronic acid and sodium hyaluronate.

The term "aqueous" refers to the fact that the majority component of the compositions of the invention is water, that is, the water is present in an amount greater than 50% by weight with respect to the total weight of the composition, preferably greater than 55%, more preferably greater than 60%, more preferably greater than 65%, more preferably greater than 70%, more preferably greater than 75%, even more preferably higher than 80%.

The hyaluronic acid present in the compositions of the present invention has an average molecular weight of from 4,000 to 20,000,000 Da, preferably from 36,000 to 13,000,000 Da, more preferably from 68,000 to 7,000,000 Da, even more preferably from 100,000 to 300,000. Gives.

The "sodium hyaluronate" or "sodium hyaluronate" is the sodium salt of hyaluronic acid, as defined above. The sodium hyaluronate present in the compositions of the present invention has an average molecular weight of 4,000 to 20,000,000 Da, preferably 336,000 to 13,800,000 Da, more preferably 668,000 to 7,600,000 Da, even more preferably 1,000. 000 to 1,400,000 Da.

In a particular embodiment, the compositions of the invention comprise hyaluronic acid of average molecular weight of 36,000 to 13,000,000 Da and sodium hyaluronate of average molecular weight of 336,000 to 13,800,000 Da, more preferably hyaluronic acid of average molecular weight of 68,000 to 7,000,000 Da and sodium hyaluronate of average molecular weight from 668,000 to 7,600,000 Da, even more preferably hyaluronic acid of average molecular weight from 100,000 to 300,000 Da and sodium hyaluronate of average molecular weight from 1,000,000 to 1,400,000 Da.

In a preferred embodiment, the compositions of the invention do not contain madecasoside. The expression "does not contain (n)", "be free of" or "are free of" means that the component to which they refer is absent or substantially absent in the compositions, for example in an amount of less than 5%, less than 4%, less than 3%, less than 2%, less than 1%, less than 0.5%, less than 0.1%, less than 0.05%, less than 0.01%, less than 0.005% , less than 0.001% or 0% by weight with respect to the total weight of the composition.

In a preferred embodiment, the weight ratio of asiaticoside to the sum of hyaluronic acid and sodium hyaluronate is from 1: 5 to 5: 1. This ratio is determined from the weight of asiaticoside present in the compositions of the invention with respect to the sum of the weight of hyaluronic acid and sodium hyaluronate in said compositions. Preferably, the weight ratio of asiaticoside to the sum of hyaluronic acid and sodium hyaluronate is from 1: 3 to 3: 1, more preferably from 1: 2 to 2: 1, still more preferably from 1.5: 1 to 1: 1.5, still more preferably from 1.1: 1 to 1: 1.1, even more preferably approximately 1: 1, most preferred 1: 1.

The term "approximately" when used to characterize a numerical value refers to a slight variation of said value, preferably ± 10% of said value, more preferably ± 5% said variation is understood by the person skilled in the art in the context in which the term is mentioned.

In another preferred embodiment, the weight ratio of sodium hyaluronate to hyaluronic acid is from 1: 5 to 5: 1, more preferably from 1: 3 to 3: 1, even more preferably from 1: 1 to 1: 3, yet more preferably from 1: 1.5 to 1: 2.5, even more preferably approximately 1: 2, most preferably 1: 2.

In another preferred embodiment, in the compositions of the invention the asiaticoside is present in an amount of 0.01% to 1% by weight relative to the total weight of the composition, preferably in an amount of 0.05% to 1%, more preferably from 0.1% to 0.5%, more preferably from 0.2% to 0.4%, more preferably from 0.25% to 0.35%, even more preferably approximately 0.3%, most preferred 0.3%.

In another preferred embodiment, in the compositions of the invention the sum of the amount of hyaluronic acid and sodium hyaluronate in the composition is 0.01% to 1% by weight relative to the total weight of the composition, preferably 0.05% to 1%, more preferably from 0.1% to 0.5%, more preferably from 0.2% to 0.4%, more preferably from 0.25% to 0.35%, even more preferably approximately 0.3%, most preferred 0.3%.

In a particular embodiment, in the compositions of the invention, hyaluronic acid is present in an amount of 0.01% to 1% by weight relative to the total weight of the composition, preferably 0.05% to 1%, more preferably 0.1% to 0.5%, more preferably from 0.1% to 0.3%, more preferably from 0.15% to 0.25%, still more preferably approximately 0.2%, most preferably 0, two%.

In another particular embodiment, in the compositions of the invention the sodium hyaluronate is present in an amount of 0.01% to 1% by weight with respect to the total weight of the composition, preferably from 0.05% to 1%, more preferably from 0.05% to 0.5%, more preferably from 0.05% to 0.2%, more preferably from 0.05% to 0.15%, still more preferably approximately 0.1%, most preferred 0 ,one%.

In another preferred embodiment, the compositions of the invention further comprise one or more wetting agents, for example 1,2, 3, 4, 5, 6, 7 or 8 wetting agents.

In the context of the present invention the term "wetting agent" refers to a compound that is capable of maintaining and retaining moisture Examples of wetting agents are polyhydric alcohols such as glycerin, propylene glycol, xylitol, sorbitol and polyethylene glycol. The humectant is selected from the group consisting of glycerin, propylene glycol, xylitol, sorbitol and mixtures thereof, even more preferably being selected from the group consisting of glycerin, propylene glycol, sorbitol and mixture thereof.

Preferably the wetting agent (s) are present in the composition in an amount of 5 to 30% by weight relative to the total weight of the composition, more preferably 10 to 30%, more preferably 15 to 30%. %, most preferred 20 to 30%.

In another preferred embodiment, the compositions of the present invention further comprise glycyrrhetinic acid or a salt thereof, preferably glycyrrhetinic acid. Glycyrrhetinic acid is also known as enoxolone or glycyrrheic acid and has the chemical structure (111) shown below.

Glycyrrhetinic acid can form salts with a base, for example, ammonium salts or inorganic salts with an alkali metal or alkaline earth metal, such as, for example, sodium, potassium, calcium, magnesium.

In a preferred embodiment, the weight ratio of glycyrrhetinic acid or a salt thereof to asiaticoside is from 1: 5 to 5: 1, more preferably from 1: 3 to 3: 1, even more preferably from 1: 1 to 1 : 3, even more preferably from 1: 1 to 1: 2, still more preferably approximately 1: 1.5, most preferred 1: 1.5.

Preferably, in the compositions of the invention glycyrrhetinic acid or a salt thereof is present in an amount of 0.01% to 1% by weight relative to the total weight of the composition, preferably in an amount of 0.05% to 1%. %, more preferably from 0.1% to 0.5%, more preferably from 0.1% to 0.3%, more preferably from 0.15% to 0.25%, even more preferably approximately 0.2%, most preferred 0.2%.

In a preferred embodiment the compositions of the invention comprise one or more film-forming agents. The term "film-forming agent" refers to compounds such as polymers or resins that form a film when applied to a surface, such as over a wound in the buccal cavity Examples of film-forming agents are carbomers, copolymers of methyl vinyl ether and maleic anhydride (PVM / MA), cellulose gums, polysaccharides, polymers and copolymers of polyvinylpyrrolidone (PVP).

Preferably the film-forming agent is selected from the group consisting of carbomer, copolymer of methyl vinyl ether and maleic anhydride (PVM / MA) and mixtures thereof.

In the context of the present invention, the term "polyvinylpyrrolidone", "povidone" or "PVP" are used interchangeably herein and refer to a polymer formed from the monomer vinylpyrrolidone. Preferably, the polyvinylpyrrolidone present in the compositions of the present invention has an average molecular weight of 20,000 to 2,000. 000 Da, preferably from 30,000 to 1,000,000, more preferably from 40,000 to 500,000.

Preferably, the PVM / MA copolymer present in the compositions of the present invention has an average molecular weight of 150,000 to 1,500,000 Da, preferably 700,000 to 1,500,000 Da.

In the context of the present invention, the term "carbomer" refers to a polymer of acrylic acid and optionally C10-C30 alkyl acrylate, crosslinked with allyl ether of pentaerythritol, allyl ether of sucrose or propyl allyl ether, preferably is a polymer of acrylic acid and C10-C30 alkyl acrylate, cross-linked with allyl ether of pentaerythritol. Preferably, the carbomer present in the compositions of the present invention has an average viscosity of 47,000 to 77,000 cP (determined with a Brookfield RVT viscometer at 20 rpm, at a carbomer concentration in water of 1% w / v (g carbomer / mL water) at pH 5.8-6.3 and 25 ° C).

In another preferred embodiment, the compositions of the present invention further comprise one or more ingredients selected from the group consisting of surfactants (such as polyoxyethylenated hydrogenated castor oil), chelating agents (such as sodium gluconate), buffering agents (as example, sodium citrate, citric acid), preservatives (such as sodium benzoate, benzoic acid, 4-isopropyl-3-methylphenol), sweeteners (such as sucralose) and flavorings.

The term "surfactant" refers to compounds that are capable of decreasing surface tension. Suitable surfactants include nonionic, anionic, cationic and amphoteric, preferably nonionic, surfactants. Examples of nonionic surfactants are ethoxylated hydrogenated castor oils (such as PEG-40 hydrogenated castor oil), block copolymers such as block copolymers of ethylene oxide and propylene oxide (poloxamers). Examples of anionic surfactants are sodium lauryl sulfate, solium lauryl sarcosinate, sodium lauryl isethionate, sodium dodecyl sulfonate, among others. Examples of cationic surfactants are lauryltrimethylammonium chloride and cetilpiridinium chloride, among others. Examples of amphoteric surfactants are myristylbetaine, palmitylbetaine, laurylbetaine, cetylbetaine, stearylbetaine, lauramidopropylbetaine, cocoamidoethylbetaine, cocoamidopropylbetaine and the like. Preferably the surfactant is polyoxyethylenated hydrogenated castor oil (such as hydrogenated castor oil PEG-40).

The term "chelating agent" refers to compounds that form complexes with metal ions Examples of chelating agents are sodium gluconate, EDTA (ethylenediaminetetraacetic acid) or its sodium salts, sodium phytate, phytic acid, potassium gluconate, sodium gluconate, acid nitrilotriacetic (NTA), ethylenediamine-N, N'-disuccinic acid (EDDS) or its sodium salts and mixtures thereof.

The term "buffering agent" refers to a compound or mixture of compounds that maintain and / or adjust the pH of the composition In particular, the pH of the composition is from 5.0 to 6.0, preferably from 5.6. to 6.0 To determine said pH the composition of the invention is dissolved in water at a concentration of 10% w / w Examples of buffering agents are sodium citrate, potassium citrate, citric acid, sodium carbonate, potassium carbonate, sodium bicarbonate, potassium bicarbonate, disodium hydrogen phosphate, dipotassium hydrogen phosphate, sodium dihydrogen phosphate, potassium dihydrogen phosphate, sodium phosphate, phosphoric acid, disodium pyrophosphate, tetrasodium pyrophosphate, sodium benzoate, benzoic acid, sodium hydroxide, potassium hydroxide , imidazole, lactic acid, sodium lactate and mixtures thereof, preferably sodium citrate, citric acid, sodium hydroxide or mixtures thereof.

The term "preservative" refers to compounds that inhibit the growth of microorganisms in the composition Examples of preservatives are sodium benzoate, benzoic acid, 4-isopropyl-3-methylphenol, dodecyltrimethylammonium bromide, tetradecylpyridinium chloride, domiphene bromide, chloride of N-tetradecyl-4-ethylpyridinium, dodecyldimethyl (2-phenoxyethyl) ammonium bromide, benzyldimethyl stearyl ammonium chloride, quaternized 5-amino-1,3-bis (2-ethylhexyl) -5-methylhexahydropyrimidine, benzalkonium chloride, benzethonium chloride, methylbenzethonium chloride and mixtures thereof, preferably sodium benzoate, benzoic acid, 4-isopropyl-3-methylphenol and mixtures thereof.

The term "sweetener" refers to a compound that imparts sweet flavor to the compositions Examples of sweeteners are sucralose, sucrose, aspartame, saccharin, stevia, steviosides, steviolglucosides, acesulfame-k, rebaudioside A, rebaudioside B, rebaudioside C, rebaudioside E, rebaudioside F, Dulcoside A, Dulcoside B, Ruboside, Neohesperidin DC, Alitame, Neotame, Thaumatin, Mucoside IV, Mosagróside V, Siamenoside, Monatin and its salts, Curculin, Monellin, Mabinlin, Brazzein, Hemandulcin, Filodulcin , glycylkylline, floridine, trilobatin, baiyanoside, oselline, polipodoside A, pterocarioside A, pterocarioside B, mukurozioside, flomyosil I, periandrine I, abrusoside A, cyclocarioside I, N- [N- [3- (3-hydroxy-4-methoxyphenyl ) propyl] -La-aspartyl] -L-phenylalanine 1-methyl ester, N- [N- [3- (3-hydroxy-4-methoxyphenyl) -3-methylbutyl] -La-aspartyl] -L-phenylalanine 1- methyl ester, N- [N- [3- (3-methoxy-4-hydroxyphenyl) propyl] -L-aspartyl] -L-phenylalanine 1-methyl ester, among others, preferably sucralose.

The term "flavorant" refers to a compound that modifies the aroma and flavor of the compositions and that can optionally serve to hide (mask) an unpleasant taste in the compositions. The flavorings can be natural or synthetic. Examples of flavorings are oils of mint, wintergreen, clove, cassia, sage, parsley, marjoram, lemon, naranga, propenyl guaetol, heliotropin, ds-4-heptenal, diacetyl, methyl-p-tert-butylphenyl acetate, methyl salicylate , ethyl salicylate, 1-mentylacetate, oxanone, airisone, methyl cinnamate, ethyl cinnamate, butyl cinnamate, ethyl butyrate, ethyl acetate, methyl anthranilate, isoamyl acetate, isoamyl butyrate, allyl caproate, eugenol Menthol, citronellol, farnesol, neohesperidin, eucalyptol, thymol, cinnamic alcohol, octanol, octanal, decanol, decanal, phenylethyl alcohol, benzyl alcohol, a-terpineol, linalool, limonene, citral, neral, geranial, geraniol, nerol, maltol, ethylmaltol, anethole, dihydroanetol, carvone, menthone, p-damascenone, ionone, Y-decalactone, Y-nonalactone, Y-undecalactone, isopulegol, piperitone and mixtures thereof.

In a particular embodiment, the composition of the invention comprises:

- from 0.05% to 1% by weight of asiaticoside,

- from 0.05% to 1% by weight of hyaluronic acid and sodium hyaluronate, and

optionally from 0.05% to 1% by weight glycyrrhetinic acid or a salt thereof, wherein the% by weight are based on the total weight of the composition.

In another particular embodiment, the composition of the invention comprises:

- from 0.1% to 0.5% by weight of asiaticoside,

- from 0.1% to 0.5% by weight of hyaluronic acid and sodium hyaluronate, and

- optionally from 0.1% to 0.5% by weight glycyrrhetinic acid or a salt thereof, wherein the% by weight are based on the total weight of the composition.

In another particular embodiment, the composition of the invention comprises:

- from 0.2% to 0.4% by weight of asiaticoside,

- from 0.2% to 0.4% by weight of hyaluronic acid and sodium hyaluronate, and

- optionally from 0.1% to 0.3% by weight glycyrrhetinic acid or a salt thereof, wherein the% by weight are based on the total weight of the composition.

In another particular embodiment, the composition of the invention comprises:

- from 0.25% to 0.35% by weight of asiaticoside,

- from 0.25% to 0.35% by weight of hyaluronic acid and sodium hyaluronate, and

- optionally from 0.15% to 0.25% by weight glycyrrhetinic acid or a salt thereof, wherein the% by weight are based on the total weight of the composition.

In another particular embodiment, the composition of the invention comprises:

- about 0.3% by weight of asiaticoside,

- about 0.3% by weight of hyaluronic acid and sodium hyaluronate, and - optionally about 0.2% by weight glycyrrhetinic acid or a salt thereof,

wherein the% by weight are relative to the total weight of the composition.

In another preferred embodiment, the composition is in the form of a topical composition, ie, suitable for topical use, for example in the form of a gel, solution, emulsion, aerosol, mousse, lotion, ointment, ointment, paste or cream, preferably in gel form.

In the context of the present invention the term "gel" refers to a colloidal system where the continuous phase is solid and the dispersed phase is liquid and does not flow when it is in a stationary state.

The compositions of the invention are preferably oral compositions, ie, suitable for oral use (in the oral cavity). This means that they do not contain (that is, they are free of) toxic components for use in the oral cavity.

The compositions of the present invention can be obtained by methods customary to the person skilled in the art, for example by combining and mixing the ingredients, preferably by the use of stirring and mixing systems, more preferably with temperature control and processing times .

Uses of the compositions

The compositions of the present invention aid in the healing of wounds and in addition have anti-inflammatory properties.

Therefore, in the second aspect the present invention relates to the compositions of the first aspect for use in medicine.

This aspect can also be formulated as the use of a composition according to the first aspect for the manufacture of a medicament.

In the third aspect, the present invention relates to the compositions of the first aspect for use in promoting wound healing.

This aspect can also be formulated as the use of a composition according to the first aspect for the manufacture of a medicament for promoting wound healing.

It can also be formulated as a method for promoting wound healing in a subject comprising administering a composition according to the first aspect.

The subject can be a human or animal, preferably a human, who has a wound. Preferably an effective amount of the composition is administered. Said therapeutically effective amount is determined by the person skilled in the art is that necessary to produce the desired effect, and generally depends on the severity of the wound to be treated, the age and / or the weight of the subject.

The composition of the present invention can be administered 1, 2, 3 or 4 times a day, preferably 1, 2 or 3 times a day, more preferably 1 or 2 times a day. Preferably the compositions of the invention are topical, in which case they can be applied over the wound.

The term "promote" refers to accelerating, helping, collaborating and improving the process of wound healing, either in time and / or quality, with respect to how it would occur in the absence of a treatment.

The term "scarring" refers to the biological process of repairing a tissue when it has suffered a wound.

The term "wound" refers to a lesion that has occurred on a tissue of the human or animal body (in particular a human), preferably on mucous membranes or skin, more preferably on mucous membranes. In a preferred embodiment, the wounds are in the buccal cavity. Such wounds may have been caused by traumatic events, such as an accident, by orthodontic appliances, by dentures, surgical interventions (such as extraction of a tooth, placement of an implant, among others); by chemotherapy or radiotherapy; for diseases such as candidiasis and oral herpes; or others. Preferably, said wounds have been caused by a surgical intervention of the oral cavity. In particular, the wounds of the oral cavity include lacerations, abrasions, penetrating wounds, sores, ulcers, among others.

The compositions of the invention have anti-inflammatory properties, so that they can also be used in the treatment and / or prevention of inflammations, in particular inflammations of tissues of the oral cavity, such as gums.

Therefore, in a fourth aspect, the present invention relates to a composition according to the first aspect for use in the treatment and / or prevention of inflammation, in particular inflammation of a tissue of the oral cavity, such as, for example, gums

This aspect can also be formulated as the use of a composition according to the first aspect for the manufacture of a medicament for the treatment and / or prevention of inflammation, in particular of inflammations of tissues of the oral cavity, such as for example gums.

It can also be formulated as a method for the treatment and / or prevention of inflammation, in particular inflammations of tissues of the oral cavity, such as gums, in a subject which comprises administering a composition according to the first aspect.

The term "treat" or "treatment" refers to reversing, alleviating, inhibiting the progress of the condition or condition to which that term applies or one or more symptoms of such a condition or condition.

The term "prevent" or "prevention" refers to the inhibition of the appearance of the condition or condition to which this term applies, or one or more symptoms of such a condition or condition.

The following non-limiting examples will further illustrate specific embodiments of the invention.

Examples

Example 1. Composition

1 Polymer of acrylic acid and C10-C30 alkyl acrylate, cross-linked with pentaerythritol allyl ether with viscosity 29400-39400 cP

1000 g of composition are prepared using the following procedure:

Water, sorbitol, carbomer and PVM / MA copolymer are mixed vigorously (using a turrax dispersing machine) for 50 min at room temperature until a homogeneous mixture is obtained. Sodium gluconate, sodium benzoate, sucralose, sodium citrate and citric acid are added to the mixture, stirred for 15 min and the pH adjusted to 5.0-5.5 with NaOH. Sodium hyaluronate, hyaluronic acid and glycerin are added while stirring is maintained. Finally, a pre-mix of hydrogenated castor oil PEG-40, propylene glycol, enoxolone, asiaticoside, benzoic acid and 4-isopropyl-3-methylphenol and aroma is added. The final mixture is stirred for another 15 min at room temperature and the pH is adjusted to 5.5-6.0 with NaOH.

Example 2. Healing test

The healing capacity of the composition obtained in Example 1 was evaluated. The test consisted of making a cut (or excision) on a cell culture of human gingival fibroblasts, simulating a wound, and checking the regeneration process (or healing). The regenerative effect of the cell culture of human gingival fibroblasts with 0 without treatment was compared.

Studies were performed on human fibroblasts from oral mucosa, specifically isolated from gingival tissue (Human Gingival Fibroblasts, HGF) from a Caucasian donor (37-year-old Australian woman, Cell Research Corporation). The HGF cells were cultured in growth medium composed of Dulbecco's culture medium 1 g / l glucose, supplemented with 10% fetal bovine serum (FBS), 2 mM L-glutamine and antibiotics (10 ^ g / ml Penicillin / 100 U / ml Streptomycin). The cells were cultured in an incubator at 37 ° C, 5% CO2 and saturated humidity conditions. For subcultivation of these cells, these were collected from the culture bottle by trypsinization with 0.25% trypsin-EDTA liquid solution (Gibco) and seeded at the cell density required for expansion or the performance of planned trials. The essays performed in this study were carried out with HGF cells at a number of subcultivations or passes less than 6. Human gingival fibroblasts (HGF) were seeded in 24-well culture plates at a density of 50,000 cells / cm2 (100,000 cells / well), in the middle of growth. These remained in growth during 48h to assure the arrival to confluence of the crop. They were then subjected to a serum deprivation process for an additional 16 - 20h. For this deprivation, the cells were maintained in assay medium. Once the deprivation was over, the wound was made on the HGF culture.

For the formation of the wound in each of the test wells, a tear of the HGF monolayer was performed vertically, by moving a single-channel pipette tip, type P1000, along the entire well ( from upper end to lower end), with an approximate angle of 30-45 ° with respect to the base of the well and with a rapid movement and constant force. In this way, a cell-free wound or area about 2 mm in amplitude was obtained. After the formation of the wound, the cultures were washed twice with HBSS buffer ( Hank's balanced salt solution) supplemented with 5 mM magnesium chloride and 10 mM calcium chloride, for the complete elimination of the cell debris produced during the performance of the wound. Next, the test solutions were applied. Three concentrations of the product were evaluated in this test, the 1/512, 1/768 and 1/1152 dilutions prepared in Dulbeco's 1 g / l glucose test medium, supplemented with 0.1% fetal bovine serum ( FBS), 2 mM L-glutamine and antibiotics (10 ^ g / ml Penicillin / 100 U / ml Streptomycin). As a negative or baseline study control, the cells were maintained in assay medium during the treatment periods. As a positive control, the cells were maintained in growth medium. As a second positive control, the effect on the healing of the growth factor Basic Fibmblast Growth Factor or bFGF (also known as FGF2 or FGF-P) prepared at a concentration of 50 ng / ml in assay medium was evaluated. Each of these conditions was evaluated in triplicate. Immediately after the application of the test solutions, initial images of the wound were obtained from each of the test wells, using an Olympus ScanR automated microscope (in a clear field, using a 4x objective). After capturing these initial images (t = 0 h), the plates were kept in incubation and treatment for 24 h and an additional 48 h. At each of these treatment times, 24 h and 48 h of treatment, new images were acquired maintaining the coordinates of each of the initial images (acquired at t = 0 h) in each assay well.

The composition of Example 1 showed a cicatrization-inducing effect of human gingival fibroblasts at the maximum concentration tested (1/512 dilution), at 24 h and 48 h after applying the composition of Example 1. The increase in healing associated with the The composition of Example 1 is 49.9% 6.561% after 24 hours of treatment, and 21.983 10.269% after 48 hours with respect to cell culture without treatment. These results are indicative of the regenerative and healing potential of the composition of example 1.

Example 3. Evaluation of the anti-inflammatory action

The antiinflammatory action of the composition obtained in Example 1 on HGF (human gingival fibroblasts) was determined by evaluating the effect of said composition on the release of various inflammatory mediators, such as cytokines, chemokines and some growth factors, in culture of HGF induced to the inflammatory response.

For this, the HGF were seeded in 24-well format culture plates at a density of 50,000 cells / cm 2 (100,000 cells / well), in the growth medium described in Example 2. They were kept growing until they reached a confluence. cellular between 80-90%. They were then subjected to a serum deprivation process for 16-20 h, using the assay medium described in Example 2. After deprivation was complete, the controls and the test product were applied.

Three concentrations of the product (composition of Example 1) were evaluated in this test, the 1/512, 1/768 and 1/1152 dilutions prepared in assay medium. As a negative or baseline control of the study, the cells were maintained in assay medium during the treatment period. As a positive control, the cells were treated with dexamethasone at a concentration of 5 ^ M. After 24h of treatment, the cultures were induced to the inflammatory response by adding LPS (lipopolysaccharide of bacterial origin) to the test medium at a concentration of 10 ^ g / ml. After 18-20h of incubation, the cell supernatants were collected and frozen (-20 ° C) for the subsequent quantification of inflammatory mediators.

The anti-inflammatory action of the product was determined by studying the effect of said product on the release of a broad panel of inflammatory mediators, in culture of HGF induced to the inflammatory response. The released levels of the inflammatory mediators IL-1a, IL-ip, IL-4, IL-6, IL-8, IL-10, IL-13, MCP-1, IFN-Y and TNF-a, were quantified in supernatants of HGF cultures induced with bacterial (+ LPS) or non-induced lipopolysaccharide (-LPS) to the inflammatory response, using the Quantibody Multiplex Human Inflammation Array Q1 system (RAYBIOTECH).

The composition of Example 1 completely inhibited the inflammatory response induced by LPS (10 ^ g / ml) in HGF culture, maintaining levels of IL-1a, IL-4, IL-8 and INF-a equal to those of the HGF not induced or not inflamed with LPS. The composition of Example 1 also exerted an important anti-inflammatory action by inhibiting the pro-inflammatory IL-6 and the chemokine MCP-1, with a maximum reduction of the released IL-6 levels of 98% and 25.45% for MPC-1, at the highest concentration tested (dilution 1/512).

Claims (24)

1. Aqueous composition comprising asiaticoside, hyaluronic acid and sodium hyaluronate. 2. The composition according to claim 1, wherein the weight ratio of asiaticoside to the sum of hyaluronic acid and sodium hyaluronate is from 1: 5 to 5: 1. The composition according to any of the preceding claims, wherein the weight ratio of sodium hyaluronate to hyaluronic acid is from 1: 5 to 5: 1. 4. The composition according to claim 1, wherein the asiaticoside is present in an amount of 0.01% to 1% by weight relative to the total weight of the composition. Composition according to any one of the preceding claims, in which the sum of the amount of hyaluronic acid and sodium hyaluronate in the composition is from 0.01% to 1% by weight with respect to the total weight of the composition. The composition according to any of the preceding claims wherein the average molecular weight of the hyaluronic acid is from 100,000 to 300,000 Da and wherein the average molecular weight of the sodium hyaluronate is from 1,000,000 to 1,400,000 Da. The composition according to any of the preceding claims, further comprising a wetting agent. The composition according to claim 7, wherein the wetting agent is selected from the group consisting of glycerin, propylene glycol, xylitol and mixture thereof. Composition according to any of claims 7 or 8, wherein the wetting agent is present in an amount of 5% to 30% by weight with respect to the total weight of the composition. The composition according to any of the preceding claims, further comprising glycyrrhetinic acid or a salt thereof. The composition according to claim 10, wherein the weight ratio of glycyrrhetinic acid or a salt thereof to asiaticoside is from 1: 5 to 5: 1. The composition according to any of claims 10 or 11, wherein the glycyrrhetinic acid or a salt thereof is present in an amount of 0.01% to 1% by weight with respect to the total weight of the composition. The composition according to any of the preceding claims, further comprising a film-forming agent. The composition according to claim 13 wherein the film-forming agent is selected from the group consisting of carbomer, methyl vinyl ether copolymer and maleic anhydride (PVM / MA) and mixtures thereof 15. The composition according to any of the preceding claims, further comprising one or more ingredients selected from the group consisting of surfactants, chelating agents, buffering agents, preservatives, sweeteners, flavorings and masking agents. 16. Composition according to any of the preceding claims in the form of a topical composition. 17. Composition according to any of the preceding claims in gel form. 18. Composition according to any of the preceding claims for use in medicine. 19. Composition according to any of claims 1 to 17 for use in promoting wound healing. 20. Composition for use according to claim 19, wherein the wounds are in the buccal cavity. 21. Composition for use according to claim 20, wherein the wounds are caused by a surgical intervention of the oral cavity. 22. The composition according to any of claims 1 to 17 for use in the treatment and / or prevention of inflammation. 23. Composition for use according to claim 22 wherein the inflammation is of a tissue of the oral cavity. 24. Composition for use according to claim 23 wherein the inflammation is of the gums.