FR2883473A1 - Topical cosmetic composition, useful to induce the tanning of the skin, to fight against cutaneous aging and to protect skin from UV rays, comprises an extract of chicle gum and a vehicle - Google Patents
Topical cosmetic composition, useful to induce the tanning of the skin, to fight against cutaneous aging and to protect skin from UV rays, comprises an extract of chicle gum and a vehicle Download PDFInfo
- Publication number
- FR2883473A1 FR2883473A1 FR0503010A FR0503010A FR2883473A1 FR 2883473 A1 FR2883473 A1 FR 2883473A1 FR 0503010 A FR0503010 A FR 0503010A FR 0503010 A FR0503010 A FR 0503010A FR 2883473 A1 FR2883473 A1 FR 2883473A1
- Authority
- FR
- France
- Prior art keywords
- cosmetic composition
- extract
- skin
- chicle gum
- chicle
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 240000001794 Manilkara zapota Species 0.000 title claims abstract description 43
- 235000011339 Manilkara zapota Nutrition 0.000 title claims abstract description 41
- 229920001412 Chicle Polymers 0.000 title claims abstract description 38
- 239000000284 extract Substances 0.000 title claims abstract description 34
- 239000000203 mixture Substances 0.000 title claims abstract description 32
- 239000002537 cosmetic Substances 0.000 title claims abstract description 21
- 230000000699 topical effect Effects 0.000 title claims abstract description 4
- 230000032683 aging Effects 0.000 title description 4
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical group CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 19
- 239000002904 solvent Substances 0.000 claims description 11
- 238000000605 extraction Methods 0.000 claims description 8
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- 125000004805 propylene group Chemical group [H]C([H])([H])C([H])([*:1])C([H])([H])[*:2] 0.000 claims 1
- 230000000694 effects Effects 0.000 abstract description 16
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Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q17/00—Barrier preparations; Preparations brought into direct contact with the skin for affording protection against external influences, e.g. sunlight, X-rays or other harmful rays, corrosive materials, bacteria or insect stings
- A61Q17/04—Topical preparations for affording protection against sunlight or other radiation; Topical sun tanning preparations
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
- A61K8/9783—Angiosperms [Magnoliophyta]
- A61K8/9789—Magnoliopsida [dicotyledons]
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
- A61Q19/04—Preparations for care of the skin for chemically tanning the skin
Landscapes
- Health & Medical Sciences (AREA)
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Abstract
Description
La présente invention concerne une composition cosmétique à base d'unThe present invention relates to a cosmetic composition based on a
extrait de gomme Chicle, utile notamment pour induire le bronzage et protéger les cellules de la peau face aux rayonnements ultraviolets. Chicle gum extract, useful in particular for inducing tanning and protecting skin cells against ultraviolet radiation.
On appelle Chicle le latex coagulé obtenu à partir du Sapotillier, Manilkara zapota syn. Manilkara achras syn. Achras sapota, arbre laticifère de la famille des Sapotacées originaire d'Amérique centrale, et plus particulièrement des forêts du Mexique, du Guatemala et du Belize. Le Chicle est obtenu après incision du tronc, écoulement du latex, récolte puis chauffage à ébullition. Chicle is the coagulated latex obtained from Sapotillier, Manilkara zapota syn. Manilkara achras syn. Achras sapota, a laticiferous tree of the family Sapotaceae native to Central America, and more particularly the forests of Mexico, Guatemala and Belize. Chicle is obtained after incision of the trunk, flow of the latex, harvest then heating to boiling.
Le Chicle est utilisé traditionnellement en Amérique centrale en tant que gomme à mâcher, d'où le nom de gomme Chicle qui lui est souvent attribué. La gomme Chicle était largement utilisée en tant que constituant de base des chewing-gums, jusqu'à l'apparition de substituts synthétiques auxquels les industriels font désormais majoritairement appel. Chicle is traditionally used in Central America as a chewing gum, hence the name Chicle gum which is often attributed to it. Chicle gum was widely used as a basic constituent of chewing gums, until the appearance of synthetic substitutes which manufacturers now mostly use.
L'utilisation de la gomme Chicle dans l'art antérieur fait uniquement référence à ses propriétés d'agent masticatoire. The use of Chicle gum in the prior art only refers to its masticatory properties.
Les inventeurs ont maintenant mis en évidence qu'un extrait de gomme Chicle présentait des propriétés cosmétiques particulièrement intéressantes. The inventors have now demonstrated that a Chicle gum extract has particularly interesting cosmetic properties.
La présente invention a donc pour objet une composition cosmétique destinée à un usage topique comprenant, outre un véhicule cosmétiquement acceptable, un extrait de gomme Chicle. The present invention therefore relates to a cosmetic composition intended for topical use comprising, in addition to a cosmetically acceptable vehicle, a Chicle gum extract.
Par véhicule cosmétiquement acceptable , on entend un véhicule adapté pour une utilisation en contact avec des cellules humaines et animales, en particulier les cellules de l'épiderme, sans toxicité, irritation, réponse allergique indue et similaire, et proportionné à un rapport avantage/risque raisonnable. By cosmetically acceptable vehicle is meant a vehicle adapted for use in contact with human and animal cells, in particular the cells of the epidermis, without toxicity, irritation, undue allergic response and the like, and proportionate to an advantage / risk ratio reasonable.
L'extrait de gomme Chicle est un extrait de latex du Sapotillier M. zapota obtenu par extraction par un solvant choisi parmi un solvant alcoolique (tel que l'éthanol), l'eau, l'acétate d'éthyle, le propylène glycol, le butylène glycol et un 30 mélange de ces solvants. Les mélanges de solvants peuvent être réalisés dans toutes les proportions possibles, l'éthanol étant le solvant préféré. Chicle gum extract is a latex extract of Sapotillier M. zapota obtained by extraction with a solvent selected from an alcoholic solvent (such as ethanol), water, ethyl acetate, propylene glycol, butylene glycol and a mixture of these solvents. The solvent mixtures can be made in all possible proportions, ethanol being the preferred solvent.
Selon un mode de réalisation préféré de l'invention, l'extrait de gomme Chicle est obtenu après broyage à température ambiante ou cryobroyage, et extraction sous agitation à température ambiante avec un solvant alcoolique, de préférence l'éthanol, pendant une à deux heures. L'extraction est réalisée avec une proportion gomme chicle broyée/solvant comprise entre 1:100 et 20:100 exprimée en poids, de préférence avec une proportion comprise entre 5:100 et 15:100 exprimée en poids. L'extrait brut obtenu est ensuite filtré, puis dilué dans l'eau à température ambiante, par exemple dans des proportions extrait brut - eau 1:1 à 1:10 exprimées en volume. Le précipité qui se forme suite à cette opération est récupéré, par exemple par centrifugation ou filtration, puis séché, par exemple par concentration sous vide, atomisation ou lyophilisation. According to a preferred embodiment of the invention, the Chicle gum extract is obtained after grinding at room temperature or cryomilling, and extraction with stirring at room temperature with an alcoholic solvent, preferably ethanol, for one to two hours. . The extraction is carried out with a milled chicle gum / solvent ratio of between 1: 100 and 20: 100 expressed by weight, preferably with a proportion of between 5: 100 and 15: 100 expressed by weight. The crude extract obtained is then filtered and then diluted with water at room temperature, for example in crude extract-water proportions 1: 1 to 1:10 expressed by volume. The precipitate which forms as a result of this operation is recovered, for example by centrifugation or filtration, and then dried, for example by concentration under vacuum, atomization or freeze-drying.
Il est à noter qu'une fraction particulière du précipité obtenu peut être purifiée ou concentrée notamment par extraction liquide/liquide au moyen de solvants organiques ou par chromatographie préparative. It should be noted that a particular fraction of the precipitate obtained can be purified or concentrated in particular by liquid / liquid extraction using organic solvents or by preparative chromatography.
L'application d'une composition cosmétique selon l'invention est effectuée par voie topique. The application of a cosmetic composition according to the invention is carried out topically.
La composition est destinée plus particulièrement au traitement de la peau, et peut se présenter sous forme d'onguent, de crème, d'huile, de lait, de pommade, de poudre, de tampon imbibé, de solution, de gel, de spray, de lotion, de suspension, de savon, ou de shampooing. The composition is intended more particularly for the treatment of the skin, and may be in the form of ointment, cream, oil, milk, ointment, powder, impregnated buffer, solution, gel, spray , lotion, suspension, soap, or shampoo.
Les compositions de l'invention peuvent être des compositions cosmétiques sous la forme d'émulsion huile dans eau ou eau dans huile, ou d'émulsion multiple, de micro-émulsion, de gel hydroalcoolique, de crème, d'huile, de lotion hydroalcoolique. The compositions of the invention may be cosmetic compositions in the form of an oil-in-water or water-in-oil emulsion, or a multiple emulsion, a microemulsion, a hydroalcoholic gel, a cream, an oil or a hydroalcoholic lotion. .
De manière préférentielle, les compositions cosmétiques selon l'invention peuvent comprendre de 0,0001 à 4 % dudit extrait exprimé en poids par rapport au poids total de la composition, de préférence de 0,01 à 1%. Preferably, the cosmetic compositions according to the invention may comprise from 0.0001 to 4% of said extract expressed by weight relative to the total weight of the composition, preferably from 0.01 to 1%.
Les compositions cosmétiques de l'invention sont particulièrement utiles pour induire le bronzage, lutter contre le vieillissement cutané, en particulier le photovieillissement et pour protéger la peau face aux rayonnements ultraviolets (UV). The cosmetic compositions of the invention are particularly useful for inducing tanning, combating skin aging, in particular photoaging, and for protecting the skin against ultraviolet (UV) radiation.
2883473 3 Les rayonnements UV peuvent induire des dommages cutanés, accélérant le vieillissement et favorisant l'apparition de cancers de la peau. 2883473 3 UV radiation can induce skin damage, accelerating aging and promoting the development of skin cancer.
Le vieillissement est un mécanisme génétiquement programmé, mais certains facteurs environnementaux l'accélèrent. Aussi, la peau prend-elle un aspect beaucoup plus âgé sur les zones exposées au soleil (visage, mains, dos) que les zones protégées. Au fil des années, l'épiderme s'atrophie et des troubles de différenciation cellulaire apparaissent (visible sous forme de croûtes plus ou moins pigmentées). Dans le derme, les cibles les plus sensibles aux UV sont les fibres élastiques qui s'attachent aux collagènes assurant la solidité et la souplesse du tissu. Les UV modifient le réseau collagénique: ils en diminuent la synthèse et en augmentent la dégradation. Ils perturbent aussi la synthèse et les propriétés de l'élastine. En plus des rayonnements directs, les radicaux libres, produits indirectement par les UV, peuvent provoquer des dégradations des fibres de collagène et d'élastine constitutives du derme. Cette altération structurale se traduit par l'apparition des rides et la diminution de l'élasticité de la peau. C'est le vieillissement photoinduit. Aging is a genetically programmed mechanism, but some environmental factors accelerate it. Also, the skin takes a much older look on areas exposed to the sun (face, hands, back) than protected areas. Over the years, the epidermis atrophies and cell differentiation disorders appear (visible in the form of crusts more or less pigmented). In the dermis, the most sensitive targets to UV are the elastic fibers that attach to collagens ensuring the strength and flexibility of the fabric. UVs modify the collagen network: they reduce its synthesis and increase its degradation. They also disrupt the synthesis and properties of elastin. In addition to direct radiation, free radicals, produced indirectly by UV, can cause degradation of collagen and elastin fibers constitutive of the dermis. This structural alteration results in the appearance of wrinkles and the decrease of the elasticity of the skin. It is the photoinduced aging.
Les carcinomes (affectant les kératinocytes) et les mélanomes (affectant les mélanocytes) sont les deux types de cancers cutanés. Ils ont pour origine un mauvais fonctionnement d'une cellule, chez laquelle les lésions induites par les UV n'ont pas été réparées. Cette cellule devient tumorale et va se développer anarchiquement en échappant à tout système de surveillance cellulaire. La croissance tumorale ( développement horizontal, ) donne lieO à une coloration caractéristique à la surface de la peau (changement d'aspect d'un grain de beauté). Des métastases peuvent résulter de cette prolifération incontrôlée ( développement vertical ) et se propager vers d'autres organes. Carcinomas (affecting keratinocytes) and melanomas (affecting melanocytes) are the two types of skin cancer. They originate from a cell malfunction, in which the UV-induced lesions have not been repaired. This cell becomes tumorous and will develop anarchically while escaping any cell monitoring system. Tumor growth (horizontal development,) gives lieO to a characteristic coloration on the surface of the skin (change of appearance of a mole). Metastases can result from this uncontrolled proliferation (vertical development) and spread to other organs.
La peau développe ses propres mécanismes de protection aux rayonnements UV. Ils peuvent être tissulaires (la mélanine dans l'épiderme) ou cellulaires (production d'antioxydants, de Heat Shock Protein, mécanismes de réparation de l'ADN) et varient en fonction du phototype. The skin develops its own protective mechanisms to UV radiation. They can be tissue (melanin in the epidermis) or cellular (production of antioxidants, Heat Shock Protein, DNA repair mechanisms) and vary depending on the phototype.
La protection la plus efficace face aux UV est la mélanine synthétisée par les mélanocytes. Ce pigment naturel est le seul filtre photostable, constant et efficace contre toutes les radiations du soleil. Chaque individu n'est pourtant pas égal devant cette synthèse puisqu'elle dépend du phototype du sujet. Par exemple les phototypes I et II ne seront jamais capables de synthétiser suffisamment de mélanine pour se protéger. The most effective protection against UV is melanin synthesized by melanocytes. This natural pigment is the only photostable filter, constant and effective against all sun radiation. Each individual is however not equal before this synthesis since it depends on the phototype of the subject. For example, phototypes I and II will never be able to synthesize enough melanin to protect themselves.
La pigmentation mélanique est un phénomène complexe dont l'acteur principal, le mélanocyte, est capable de synthétiser un filtre solaire protecteur naturel, très efficace (Pathak et al 1995) et piégeur de radicaux libres. Melanic pigmentation is a complex phenomenon of which the main actor, the melanocyte, is able to synthesize a natural protective filter, very effective (Pathak et al 1995) and scavenger of free radicals.
Stimulés par les UV, les kératinocytes produisent une hormone destinée aux mélanocytes, l'aMSH (a melanocyte stimulating hormone). Les mélanocytes peuvent aussi être stimulés par le FGF (fibroblast growth factor), le NGF (neuronal growth factor), le TGFa (transforming growth factor a) ou le beta-oestradiol...11s déclenchent alors la biosynthèse de mélanine à partir de la tyrosine et de l'enzyme clé de la mélanogenèse, la tyrosinase. Il existe deux types de mélanine: phéomélanine (pigment jaune-rouge) et eumélanine (pigment noir-brun), issues de deux voies différentes impliquant de nombreuses enzymes. Les mélanines synthétisées sont stockées dans les mélanosomes et transférées dans les kératinocytes adjacents par les dendrites des mélanocytes. Les mélanines jouent alors un rôle protecteur du noyau des cellules épidermiques. Les kératinocytes, ainsi pigmentés, entament leur processus de différenciation jusqu'à la surface de la peau. Stimulated by UV, keratinocytes produce a hormone for melanocytes, aMSH (a melanocyte stimulating hormone). Melanocytes can also be stimulated by FGF (fibroblast growth factor), NGF (neuronal growth factor), TGFα (transforming growth factor a) or beta-estradiol ... 11s then trigger the melanin biosynthesis from the tyrosine and the key enzyme of melanogenesis, tyrosinase. There are two types of melanin: pheomelanin (yellow-red pigment) and eumelanin (black-brown pigment), from two different pathways involving many enzymes. The synthesized melanins are stored in melanosomes and transferred to adjacent keratinocytes by melanocyte dendrites. Melanins then play a protective role of the epidermal cell nucleus. The keratinocytes, thus pigmented, begin their process of differentiation to the surface of the skin.
Ces kératinocytes sont destinés, dans des délais proches, à desquamer, favorisant ainsi le renouvellement de l'épiderme. La desquamation est un phénomène normal et ultime de la différenciation épidermique, assuré par la rupture des liaisons intercornéocytaires et du remaniement des lipides cutanés. These keratinocytes are intended, in a short time, to desquamate, thereby promoting the renewal of the epidermis. The desquamation is a normal and ultimate phenomenon of the epidermal differentiation, ensured by the rupture of the intercorneocyte bonds and the reworking of the cutaneous lipids.
Les compositions cosmétiques comprenant l'extrait de gomme Chicle sont capables d'induire de manière notable la synthèse de mélanine par les mélanocytes. Elles diminuent en outre la différenciation des kératinocytes, ce qui prolonge la durée de présence de la mélanine produite. En effet, le ralentissement de la différenciation des kératinocytes entraîne le ralentissement de la desquamation. A la surface de l'épiderme les kératinocytes disposent d'autant plus longtemps de mélanines pour assurer leur protection face aux agressions des UV. Cosmetic compositions comprising Chicle gum extract are capable of significantly inducing melanin synthesis by melanocytes. They also reduce the differentiation of keratinocytes, which prolongs the duration of presence of the melanin produced. Indeed, the slowing down of the differentiation of the keratinocytes causes slowing down of the desquamation. On the surface of the epidermis keratinocytes have longer melanins for their protection against UV aggressions.
2883473 5 Par ces mêmes propriétés, les compositions cosmétiques comprenant l'extrait de gomme Chicle induisent le bronzage, et sont utiles pour lutter contre le vieillissement cutané et protéger la peau face aux rayonnements UV. By these same properties, the cosmetic compositions comprising Chicle gum extract induce tanning, and are useful for combating skin aging and protecting the skin against UV radiation.
EXEMPLE 1EXAMPLE 1
Préparation d'un extrait de gomme Chicle. Preparation of a Chicle gum extract.
Dans les exemples ci-dessous, on utilise comme extrait de gomme Chicle, 1 o un extrait préparé de la façon suivante. La gomme Chicle sous sa forme native est broyée à température ambiante et extraite par l'éthanol 96% sous agitation à température ambiante, dans une proportion gomme chicle broyée éthanol 15:100 exprimée en poids. L'extraction est réalisée pendant 1 heure. L'extrait brut obtenu est ensuite filtré avec un seuil de coupure final de 5 pm, puis dilué dans l'eau à température ambiante dans la proportion extrait brut eau 1:1 exprimée en volumes. Le précipité formé est récupéré par centrifugation puis séché par évaporation sous vide. In the examples below, as Chicle gum extract, an extract prepared in the following manner is used. The Chicle gum in its native form is milled at ambient temperature and extracted with 96% ethanol with stirring at room temperature, in a ratio of 15: 100 ethanol milled chicom gum expressed by weight. The extraction is carried out for 1 hour. The crude extract obtained is then filtered with a final cut-off of 5 .mu.m and then diluted with water at room temperature in the ratio of crude extract water 1: 1 expressed in volumes. The precipitate formed is recovered by centrifugation and then dried by evaporation under vacuum.
Cette extraction permet d'obtenir un extrait sec dont l'efficacité a été testée vis à vis de différentes propriétés cosmétiques. This extraction makes it possible to obtain a dry extract whose effectiveness has been tested with respect to various cosmetic properties.
EXEMPLE 2EXAMPLE 2
Auqmentation de la synthèse de mélanine dans une culture de mélanocytes épidermiques humains normaux A) Matériels et méthodes Les mélanocytes épidermiques humains normaux (NEHM) sont ensemencés dans des plaques 24 puits à raison de 120 000 cellules/puit et incubés dans un incubateur à 37 C, 5% de CO2 et 95% d'humidité. Le milieu de culture est composé du milieu Melanocyte Growth Medium (MGM) M2 complémenté (Promocell C-24300), ainsi que 50UI/mL de pénicilline (Invitrogen, réf. 15140122) et 50pgImL de streptomycine (Invitrogen 15070063). Des plaques supplémentaires sont réalisées afin d'évaluer la viabilité cellulaire. Enhancing Melanin Synthesis in a Normal Human Epidermal Melanocyte Culture A) Materials and Methods Normal human epidermal melanocytes (NEHMs) are seeded in 24-well plates at 120,000 cells / well and incubated in a 37 C incubator. , 5% CO2 and 95% humidity. The culture medium is composed of complemented Melanocyte Growth Medium (MGM) M2 medium (Promocell C-24300), as well as 50 IU / ml of penicillin (Invitrogen, ref 15140122) and 50 μg / ml of streptomycin (Invitrogen 15070063). Additional plates are made to evaluate cell viability.
Les NHEM sont traités pendant 10 jours avec l'extrait de gomme Chicle solubilisé puis dilué dans le milieu de culture à 0,3pg/mL et 0, 06pg/mL. Un témoin milieu de culture est également réalisé. Afin de valider l'étude, un témoin positif, l'isobutylmethyl xanthine (IBMX, Sigma, 17018) , stimulateur de la synthèse des mélanines, est testé dans les mêmes conditions à la concentration de 200pM, ainsi qu'un témoin négatif, l'acide kojique (Sigma, K3125), inhibiteur de la synthèse des mélanines, testé aux concentrations de 80pg/mL et 35pglmL. Les différents milieux et traitements sont renouvelés tous les 3 jours. The NHEMs are treated for 10 days with the solubilized Chicle gum extract and then diluted in the culture medium to 0.3 μg / ml and 0.06 μg / ml. A culture medium control is also performed. In order to validate the study, a positive control, isobutylmethyl xanthine (IBMX, Sigma, 17018), stimulator of melanin synthesis, is tested under the same conditions at a concentration of 200 μM, as well as a negative control, kojic acid (Sigma, K3125), a melanin synthesis inhibitor, tested at concentrations of 80 μg / ml and 35 μg / ml. The different environments and treatments are renewed every 3 days.
Les milieux de culture sont éliminés à la fin du traitement et la mélanine est extraite par une solution de NaOH à 0,5N. La densité optique est lue à 405nm, contre une gamme de mélanine exogène (0,39 à 100 pg/ml) (Sigma, M8631). The culture media are removed at the end of the treatment and the melanin is extracted with 0.5N NaOH solution. The optical density is read at 405 nm against an exogenous melanin range (0.39 to 100 μg / ml) (Sigma, M8631).
La viabilité des cellules est évaluée par un test au MTT (Methylthiazoletetrazolium) et la quantité de protéines est évaluée à l'aide du kit de dosage "DC Protein Assay" (BioRad, 500-0116). La prise en compte de la quantité de protéines cellulaires dans l'expression des résultats permet de tenir compte de l'activité métabolique des cellules. The viability of the cells is evaluated by an MTT test (Methylthiazoletetrazolium) and the amount of protein is evaluated using the "DC Protein Assay" assay kit (BioRad, 500-0116). Taking into account the amount of cellular proteins in the expression of the results makes it possible to take into account the metabolic activity of the cells.
B) Résultats L'IBMX, testé à 200pM permet d'augmenter significativement la quantité de mélanine synthétisée de 34 % par rapport au témoin non traité. Cet effet est corrélé avec une augmentation de la quantité de protéines présentes de 33% et une stimulation de la prolifération cellulaire de 39%. B) Results The IBMX, tested at 200 μM makes it possible to significantly increase the amount of melanin synthesized by 34% compared to the untreated control. This effect is correlated with an increase in the amount of protein present of 33% and a stimulation of cell proliferation of 39%.
L'acide kojique provoque une inhibition de la synthèse de mélanine de façon dose dépendante. Cet effet est significatif aux deux concentrations testées, que la quantité de mélanine soit rapportée ou non à la quantité de protéines des 1 o cultures, et cela sans réduction de la viabilité cellulaire. Kojic acid causes inhibition of melanin synthesis in a dose-dependent manner. This effect is significant at the two concentrations tested, regardless of whether the amount of melanin is related to the amount of protein of the 1 0 cultures, without reducing the cell viability.
L'extrait de gomme Chicle testé aux concentrations de 0,3 pg/mL et 0,6 pg/mL permet d'augmenter très fortement la quantité de mélanine présente dans les cultures de mélanocytes, de 156% et 71% respectivement par rapport au milieu de culture seul, considéré comme le témoin non traité. Chicle gum extract tested at concentrations of 0.3 μg / ml and 0.6 μg / ml makes it possible to increase very considerably the amount of melanin present in the melanocyte cultures by 156% and 71% respectively relative to culture medium alone, considered the untreated control.
Cet effet s'accompagne d'une nette augmentation de la quantité de protéines mélanocytaires. Cependant l'effet stimulant de l'extrait de gomme Chicle reste significatif lorsque la quantité de mélanine synthétisée est rapportée à la quantité de protéines présentes, ceci afin de prendre en compte l'activité métabolique des cellules. This effect is accompanied by a marked increase in the amount of melanocyte proteins. However, the stimulating effect of Chicle gum extract remains significant when the amount of melanin synthesized is related to the amount of protein present, in order to take into account the metabolic activity of the cells.
Tableau 1: effet de l'extrait de gomme Chicle sur la synthèse de mélanine dans une culture de NHEM Traitement Concentration Mélanine Mélanine Protéines MTT (%) testée (pglmL) (% par rapport (mg/mL) au témoin) Témoin - 28,6 100 0,65 100 IBMX 200pM 38,5 134 0,86 139 Acide kojique 80pg/mL 19, 7 69 0,58 90 35pg/ml 21,6 76 0,57 92 Extrait sec de 0, 3pg/ml 73,2 256 0, 93 115 gomme Chicle 0,06pg/ml 48,8 171 0,82 109 Table 1: Effect of Chicle Gum Extract on Melanin Synthesis in a Culture of NHEM Treatment Concentration Melanin Melanine Protein MTT (%) Tested (pglmL) (% versus (mg / mL) to control) Control - 28, 6 100 0.65 100 IBMX 200 μM 38.5 134 0.86 139 Kojic acid 80 μg / ml 19.7 69 0.58 90 35 μg / ml 21.6 76 0.57 92 Dry extract of 0.3 μg / ml 73 2,256 0, 93,115 Chicle gum 0.06 g / ml 48.8 171 0.82 109
Exemple 3Example 3
Différenciation des kératinocytes humains en culture monocouche A) Matériels et méthodes Les kératinocytes épidermiques humains normaux (NHEK) R3K015 sont ensemencés à 10000 cellules/puits dans des plaques 96 puits et précultivés en milieu SFM1x (Serum Free Medium). Après incubation pendant 24 heures à 37 C, le milieu de culture est éliminé et remplacé par du milieu SFM1x contenant ou non (témoin) l'extrait de gomme Chicle ou les références acide rétinoïque (Sigma R2625, 10-7M), rétinol (Sigma, R7632, 10-6M) et le chlorure de calcium (Prolabo, 22317297, 1,5mM) . Après 48 heures d'incubation à 37 C et 5% CO2, le milieu de culture est éliminé et les cellules sont à nouveau traitées puis incubées pendant 48 heures à 37 C. Toutes les conditions expérimentales ont été réalisées en hexaplicate. Differentiation of human keratinocytes in monolayer culture A) Materials and methods Normal human epidermal keratinocytes (NHEK) R3K015 are inoculated at 10,000 cells / well in 96-well plates and precultured in SFM1x medium (Serum Free Medium). After incubation for 24 hours at 37 ° C., the culture medium is removed and replaced with SFM1x medium containing or not (control) Chicle gum extract or retinoic acid (Sigma R2625, 10-7M), retinol (Sigma , R7632, 10-6M) and calcium chloride (Prolabo, 22317297, 1.5mM). After 48 hours of incubation at 37 ° C. and 5% CO 2, the culture medium is removed and the cells are again treated and then incubated for 48 hours at 37 ° C. All the experimental conditions were carried out in hexaplicate.
Les tapis cellulaires sont lavés puis soniqués, sur glace, en tampon Tris/EDTA, à pH 8,0. L'enzyme TGk (transglutaminase kératinocytaire) membranaire est extraite en présence de triton X-100 (incubation sous agitation à 4 C), puis l'activité TGk est déterminée en mesurant l'addition covalente de putrescine tritiée à 2pCi/ml (Amersham TRK 414; 1, 55TBq/mmol, 42 Ci/mmole) à la caséine (protéine acceptrice, 2mg/ml), dans un tampon Tris 0,IM, CaCl2 4mM, EDTA 0,4mM, DTT 5mM, pH 8,3, pendant 2 heures à 37 C. The cell mats are washed and then sonicated, on ice, in Tris / EDTA buffer, at pH 8.0. Membrane TGK (keratinocyte transglutaminase) enzyme is extracted in the presence of triton X-100 (incubation with stirring at 4 ° C.), then the TGk activity is determined by measuring the covalent addition of tritiated putrescine at 2 μCi / ml (Amersham TRK 414; 1, 55TBq / mmol, 42 Ci / mmol) to casein (acceptor protein, 2mg / ml), in 0.1M Tris buffer, 4mM CaCl2, 0.4mM EDTA, 5mM DTT, pH 8.3, 2 hours at 37 degrees C.
La caséine est précipitée par addition d'acide trichloroacétique (TCA) 20% contenant 1mM de putrescine et les précipités sont récupérés sur filtres et collecteur Skatron (lavages extensifs en TCA 5%, 0,1 mM putrescine et éthanol) puis comptage des filtres secs en scintillation liquide. The casein is precipitated by the addition of 20% trichloroacetic acid (TCA) containing 1 mM of putrescine and the precipitates are recovered on filters and Skatron collector (extensive washes in 5% TCA, 0.1 mM putrescine and ethanol) and counting of the dry filters. in liquid scintillation.
Les protéines de chaque échantillon sont dosées à l'aide du kit Dc Protein Assay (BioRad 500-0116) et selon le protocole préconisé par le fournisseur. La prise en compte de la quantité de protéines cellulaires dans l'expression des résultats permet de tenir compte de l'activité métabolique des cellules. The proteins of each sample are assayed using the Dc Protein Assay kit (BioRad 500-0116) and according to the protocol recommended by the supplier. Taking into account the amount of cellular proteins in the expression of the results makes it possible to take into account the metabolic activity of the cells.
B) Résultats Les données brutes de comptage sont transférées et traitées sous le logiciel PRISM (Graph Pad Software). Les comparaisons intergroupes sont réalisées par analyse de variance (ANOVA) à l'aide du test de comparaison multiple Dunnett. B) Results The raw count data are transferred and processed under the software PRISM (Graph Pad Software). Intergroup comparisons are performed by ANOVA using the Dunnett Multiple Comparison Test.
Les résultats du dosage permettent d'observer une activité de l'ordre de quatre fois plus importante lors de l'incorporation de calcium dans le milieu de culture En effet celui-ci présente une activité inductrice connue de la différenciation des NHEK. The results of the assay make it possible to observe an activity of the order of four times greater during the incorporation of calcium into the culture medium. Indeed, this has a known inducing activity for the differentiation of NHEKs.
L'acide rétinoïque à une concentration de 10-7M et le rétinol à une concentration de 10-6M provoquent une réduction de l'activité de la TGk de 58% et 64% respectivement. Retinoic acid at a concentration of 10-7M and retinol at a concentration of 10-6M resulted in a reduction in TGk activity of 58% and 64% respectively.
L'extrait de gomme Chicle, aux concentrations de 2pg/mI et 0, 08pglml permet de réduire la quantité de TGk active de 18% et 29 % respectivement. Chicle gum extract, at concentrations of 2 μg / ml and 0.08 μg / ml, reduces the amount of active TGk by 18% and 29%, respectively.
Des résultats comparables sont obtenus lorsque l'on reporte l'activité de la TGk à la quantité de protéines kératinocytaires présentes afin de prendre en compte l'activité métabolique des cellules. Dans ce cas l'activité de l'extrait de gomme Chicle aux concentrations 2pg/ml et 0, 08pg/mI est de 23% et 24% respectivement. Comparable results are obtained when the activity of TGk is compared with the amount of keratinocyte proteins present in order to take into account the metabolic activity of the cells. In this case, the activity of chicle gum extract at concentrations of 2 μg / ml and 0.08 μg / ml is 23% and 24%, respectively.
L'extrait de gomme Chicle testé à 2pg/mL et 0,08pg/mL provoque une diminution de l'activité de la transglutaminase kératinocytaire. Il ralentit la différenciation des kératinocytes. Chicle gum extract tested at 2 μg / ml and 0.08 μg / ml caused a decrease in the activity of keratinocyte transglutaminase. It slows the differentiation of keratinocytes.
Parallèlement à ces résultats, il a été démontré que l'extrait de gomme Chicle aux concentrations 2pg/mL et 0,08pg/mL ne présente aucun effet sur la prolifération des kératinocytes, donc sur leur croissance. In parallel with these results, Chicle gum extract at concentrations of 2 μg / ml and 0.08 μg / ml has been shown to have no effect on the proliferation of keratinocytes, and therefore on their growth.
Tableau 2: effet de l'extrait de gomme Chicle sur la différenciation des kératinocytes humains en culture. Table 2: effect of Chicle gum extract on the differentiation of human keratinocytes in culture.
Traitement Concentration Activité TGk Activité TGk Protéines Activité / Activité / testée (cpm) (% par rapport (pg) protéines protéines au témoin) (% par rapport au témoin) Témoin _ 1474 100 4,4 336 100 (sans Cal+) Témoin 1,5 mM 5090 345 5,6 914 272 (avec Cal+) Rétinol 10-5M 533 36 3,7 146 43 Acide rétinoïque 10-' M 620 42 3,8 162 48 Extrait sec de 2 pg/ml 1214 82 4,7 259 77 gomme Chicle 0,08 pg/mI 1053 71 4,2 254 76 EXEMPLE 6: Formulations 1 o Exemple de composition sous forme d'émulsion (% en poids de la composition) Eau: QSP Extrait sec de gomme Chicle: 0,1 % Mélange de conservateurs: 1,50 % Propylène glycol: 5,00 % Gomme xanthane: 0,30 % Copolymère acrylique / acrylate: 0,50 % Acide stéarique polyéthoxylé 100 OE: 3,00 % Stéarate de sorbitan: 2,00 % Sorbitan lauréate polyéthoxylé 20 OE: 3,00 2 0 Alcool Cétylstéarique: 1, 50 0/0 Cire d'abeille: 1,00 cYo Huile de germe de blé : 5,00 % Diméthicone: 2,00 % Cyclométhicone: 5,00 Gel de polyacrylamide: 2,00 % Parfum: 0,10% Eau: QSP Extrait sec de gomme Chicle: 0,1 % Gomme xanthane: 0,30 0/0 Séquestrants: 0,05 Conservateurs: 1,50 % Acide palmitique: 2,50 % Acide stéarique: 2,50 % Trilaurine: 1,00 % Beurre de karité : 0,05 % Acétate de tocophérol: 0,05 % Bêta bisabolol: 0,05 % Huile de germe de blé : 5,00 % Diméthicone: 2,00 % Acide polyacrylique: 1,50 0/0 TEA (triéthanolamine) : 1,50 0/0 Parfum: 0,10 % Exemple de composition sous forme de crème (% en poids de la composition) 1 o 20 2883473 12 Treatment Concentration Activity TGk Activity TGk Protein Activity / Activity / Tested (cpm) (% relative (pg) Protein Protein to Control) (% relative to control) Control 1474 100 4.4 336 100 (without Cal +) Control 1, 5 mM 5090 345 5.6 914 272 (with Cal +) Retinol 10-5M 533 36 3.7 146 43 Retinoic acid 10- 'M 620 42 3.8 162 48 Dry extract of 2 μg / ml 1214 82 4.7 259 77 gum Chicle 0.08 g / ml 1053 71 4.2 254 76 EXAMPLE 6 Formulations 1 Example of composition in emulsion form (% by weight of the composition) Water: QSP Dry gum extract Chicle: 0.1 % Blend of preservatives: 1,50% Propylene glycol: 5,00% Xanthan gum: 0,30% Copolymer acrylic / acrylate: 0,50% Polyethoxylated stearic acid 100 EO: 3,00% Sorbitan stearate: 2,00% Sorbitan polyethoxylated laureate 20 EO: 3.00 2 0 Cetylstearic alcohol: 1, 50 0/0 Beeswax: 1.00 cYo Wheat germ oil: 5.00% Dimethicone: 2.00% Cyclomethicone: 5.00 Polyacrylamide gel: 2.00% Fragrance: 0.10% Water: QSP Dry gum extract Chicle: 0.1% Xanthan gum: 0.30 0/0 Sequestering agents: 0.05 Preservatives: 1.50% Palmitic acid: 2.50% Stearic acid: 2 , 50% Trilaurine: 1.00% Shea butter: 0.05% Tocopherol acetate: 0.05% Bisabolol: 0.05% Wheat germ oil: 5.00% Dimethicone: 2.00% Polyacrylic acid : 1.50% TEA (triethanolamine): 1.50% Fragrance: 0.10% Example of composition in the form of cream (% by weight of the composition) 1 o 20 2883473 12
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| DE4426148A1 (en) * | 1994-07-22 | 1996-01-25 | Henkel Kgaa | Cosmetic and / or pharmaceutical preparations |
| FR2818141A1 (en) * | 2000-12-14 | 2002-06-21 | Sederma Sa | COSMETIC OR DERMOPHARMACEUTICAL COMPOSITIONS CONTAINING VEGETABLE EXTRACTS OF BELLE DE NUIT (MIRABILIS JALAPA) AGAINST INFLAMMATION AND SKIN DROUGHT OF ANY ORIGIN |
| WO2004017934A1 (en) * | 2002-08-16 | 2004-03-04 | Coty B.V. | Skin-smoothing cosmetic based on plant extracts |
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| FR2609397A1 (en) * | 1988-02-23 | 1988-07-15 | Serobiologiques Lab Sa | Use of a substance or composition of carbohydrate nature as active principle of a dermatological and/or cosmetological and/or pharmaceutical and/or cell-stimulating composition, and composition containing such a substance or composition of carbohydrate nature |
| DE4426148A1 (en) * | 1994-07-22 | 1996-01-25 | Henkel Kgaa | Cosmetic and / or pharmaceutical preparations |
| FR2818141A1 (en) * | 2000-12-14 | 2002-06-21 | Sederma Sa | COSMETIC OR DERMOPHARMACEUTICAL COMPOSITIONS CONTAINING VEGETABLE EXTRACTS OF BELLE DE NUIT (MIRABILIS JALAPA) AGAINST INFLAMMATION AND SKIN DROUGHT OF ANY ORIGIN |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| WO2010041273A2 (en) | 2008-10-03 | 2010-04-15 | Rubicon Research Private Limited | Compositions comprising fenugreek hydrocolloids |
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