Classifications

• • 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
  • A61K8/72—Cosmetics or similar toiletry preparations characterised by the composition containing organic macromolecular compounds
  • A61K8/73—Polysaccharides
• • 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
  • A61K8/9706—Algae
  • A61K8/9717—Rhodophycota or Rhodophyta [red algae], e.g. Porphyra
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P17/00—Drugs for dermatological disorders
• • 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
  • A61Q19/08—Anti-ageing preparations

Definitions

• the present invention relates to the use of an active ingredient derived from Eucheuma cottonii rich in specific molecules, to combat aging of the skin and improve the age perceived, as well as a particular active ingredient containing such molecules.
• the invention also relates to cosmetic compositions including this active ingredient, and a cosmetic treatment method for preventing and / or combating cutaneous aging.
• senescence At the cellular level, aging, called senescence, is an irreversible state in which damaged cells no longer proliferate.
• the latter is inherent in cell division and signifies the decline of the replicative potential of fibroblasts.
• the cells are in fact equipped with a division quota which decides their lifetime. When this quota is exhausted, the fibroblasts stop their proliferation and enter senescence, a normal physiological process that avoids the transformation of the cell.
• telomeres Non-coding, highly-repeated DNA sequences located at the ends of chromosomes, and developed by a protein complex specific to the telosome. Over the course of cellular divisions, telomeres progressively shorten to a minimum critical size that triggers the state of cellular senescence, contributing to degeneration of the skin tissue and aging.
• the objective of the present invention is to provide a cosmetic active ingredient that promotes the protection of the telosome and telomeres of cutaneous cells, to maintain the cell replicative potential and improve the perceived age.
• the invention aims at the use of particular molecules obtained from Eucheuma coffonii.
• Eucheuma cottonii is a red, branched, green to orange-yellow algae grown mainly in Asia, particularly in the Pacific zone for the production of gelling and thickening agents used in the food, pharmaceutical and cosmetic sectors.
• the application FR2480138 describes a sulphated extract Eucheuma cottonii as a viscous food additive.
• Extracts have also been used in cosmetics for moisturizing, emulsifying, emollient, smoothing, gelling or slimming properties.
• the application FR2719846 is known which mentions the use of sulphated galactans having a sulphatation rate of at least 0.5 sulphate per saccharide unit in order to improve the elasticity of the skin and to promote healing.
• These sulphated galactans can be carrageenans obtained from Eucheuma sp., By a process involving fermentation by a marine bacterium.
• JP2002193736 which describes aqueous extracts of Eucheuma cottonii for smoothing the skin and hair.
• the present invention specifically relates to the use of an active ingredient obtained by hydrolysis of Eucheuma cottonii containing linear galactans, as a cosmetic active ingredient in a cutaneous application composition said active ingredient and / or said composition being intended (s) to promote the protection of the telosome and telomere in skin cells.
• the active ingredient contains very little if any sulfated galactans.
• the invention also relates to a particular active ingredient, namely an Eucheuma cottonii hydrolyzate comprising linear galactans, intended to be incorporated into a cutaneous application composition.
• cutaneous application composition any composition intended to be applied to the skin, preferably a cosmetic composition.
• hydrolyzate any extract obtained from Eucheuma cottonii, comprising at least one hydrolysis step.
• the subject of the invention is also a cosmetic composition containing an active ingredient obtained from Eucheuma cottonii containing linear galactans, as well as a cosmetic treatment method intended to prevent and / or fight against the effects of age on the skin, comprising the topical application to the skin of such a composition.
• the invention relates to a hydrolyzate of Eucheuma cottonii comprising linear galactans, for its use as a cosmetic active ingredient in a cutaneous application composition, said active ingredient and / or said composition being intended to promote the protection of the telosome and telomere cells of the skin.
• Telomeres guarantee the integrity of chromosomes. They protect the ends of chromosomes on the one hand to prevent them from degrading, and they also prevent the loss of genetic information during replication. However, these chromosome ends are fragile and are themselves stabilized by a multi-protein complex called telosome or shelterin.
• telosome plays a very important role of protection by allowing the telomeres to adopt a particular conformation in lasso, called telomeric loop. It masks the telomeres and thus preserves degradations by allowing however their access at regular intervals to allow their replication.
• the telosome is composed of 6 central proteins, including POT1 ("Protection Of Telomeres” protection of telomeres) and TPP1 ("POT1 and TTN2-interacting protein” proteins interacting with POT1 and TTN2). These telosome proteins participate actively in the protection of telomeres and the Applicant has notably demonstrated that their expressions are significantly reduced in senescent fibroblasts in comparison with normal fibroblasts.
• telomeres When telosomes are altered or in the absence of telosomes, weakened telomeres shorten faster at each division. Stress (UV, reactive oxygen species, etc.) also increases their wear.
• telomeres The alteration of telomeres, resulting both from their gradual erosion and / or their unmasking via telosomal failure, controls the arrest of proliferation and leads to cellular senescence, called replicative or telomeric senescence. This process involves, through various mechanisms, a gradual decline in the functionality of the skin tissue and its aging.
• an active ingredient obtained from Eucheuma cottonii comprising linear galactans makes it possible to limit the losses in the telomeres of fibroblasts.
• an active ingredient obtained from Eucheuma cottonii comprising linear galactans can be used for:
• telosomal proteins whose expressions are reduced in senescerous fibroblasts
• the active principle according to the invention makes it possible to preserve the replicative capital of the fibroblasts and to delay the phenomena of senescence and degeneration.
• the invention relates to the use in a composition of an active ingredient obtained from Eucheuma cottonii, as described below.
• the invention also relates to a particular cosmetic active ingredient, namely a hydrolyzate with! Eucheuma cottonii comprising linear galactans.
• the degree of polymerization of the linear galactans is preferably between 6 and 18.
• the linear galactans useful according to the invention are not linear sulphated galactans: less than 1% of the linear galactans present in the hydrolyzate according to the invention are sulphated.
• the active ingredient preferably has a light yellow color.
• non-viscous liquid product It is in the form of a non-viscous liquid product. It can be defined by at least one of the features described below, preferably all.
• the dry matter content of an active ingredient according to the invention (measured by passing in an oven at 105 ° C. in the presence of sand of a sample of initial weight given until a constant weight is obtained) can be between 30 and 100 g / l, preferably between 47 and 65 g / l.
• the pH (measured by the potentiometric method at room temperature) can be between 3 and 5, preferably between 3 and 4.
• the determination of the total sugar content can be carried out by the DUBOIS method (DUBOIS M. et al., (1956), Analytical Chemistry, 28, No. 3, pp. 350-356). In the presence of concentrated sulfuric acid and phenol, the reducing sugars give an orange-yellow compound. From a standard range, the total sugar content of a sample can be determined.
• the total sugar content may be greater than 45% by weight of dry matter, preferably greater than 60%.
• the total sugar content is between 19 and 71 g / l, in particular between 31 and 46 g / l.
• the determination of the carbohydrate size of an active ingredient according to the invention is carried out by high performance liquid chromatography.
• the chromatogram obtained shows the presence of monosaccharides of molecular weight less than 180 Da and oligosaccharides and polysaccharides with a molar mass of between 180 and 8100 Da (degree of polymerization at most 45).
• the monosaccharides represent in less than 35%, preferably between 18 and 25%, and oligosaccharides and polysaccharides with a molar mass of between 180 and 8100 Da represent at least 65%, preferably between 72 and 85%.
• the carbohydrate fraction of the active principle according to the invention is therefore essentially composed of monosaccharides, oligosaccharides and polysaccharides with a degree of polymerization of less than 45.
• the majority of the carbohydrates present in the active principle are linear galactans.
• the level of sulfated carbohydrates in aqueous matrices is determined using a spectrophotometric method.
• Sulphated carbohydrates give a blue color in the presence of azure A.
• the level of sulphated carbohydrates in the samples is determined relative to a standard range made with a sulfated dextran solution of known concentration.
• the raw ash content is determined by the weighing of residues from incineration at 550 ° C in an electric muffle furnace (VULCAN TM). The weight of the residue is calculated by deducting the tare.
• the mineral content is expressed as a percentage relative to the total weight of the dry matter of the active ingredient.
• the raw ash content of an active ingredient according to the invention is preferably less than 30%.
• the intensity of staining is proportional to the amount of uronic acids.
• the readings are made from a standard range of galacturonic acid ranging from 10 to 100 mg / l.
• Samples of the active ingredient according to the invention must be previously diluted with distilled water.
• the uronic acid content of the products is then directly read on this line.
• the uronic acid content of an active ingredient according to the invention is preferably less than 25%.
• the active ingredient according to the invention as described above may be preferably by a process comprising a hydrolysis.
• a particularly suitable method comprises at least the succession of the following steps:
• the process comprises at least the following succession of steps:
• the parameters of the different steps must be adjusted in order to obtain active principles comprising linear galactans, preferably linear galactans with a degree of polymerization mainly between 6 and 18.
• the production process according to the invention in particular the hydrolysis of the carbohydrates carried out, is carried out in order to eliminate the sulphated carbohydrates naturally present in the algae.
• the present invention also covers cosmetic compositions including linear galactans obtained from Eucheuma cottonii or an Eucheuma cottonii hydrolyzate comprising linear galactans, in different galenic forms, adapted for topical administration to the skin.
• compositions may in particular be in the form of oil-in-water emulsions, water-in-oil emulsions, multiple emulsions (Water / Oil / Water or Oil / Water / Oil) which may be optionally microemulsions or nanoemulsions, or in the form of solutions, suspensions, hydrodispersions, aqueous gels or powders. They can be more or less fluid and have the appearance of a cream, a lotion, a milk, a serum, an ointment, a gel, a paste or a foam, or in solid form.
• compositions comprising between 0.01 and 3% of principle (s) obtained from Eucheuma cottonii comprising linear galactans according to the present invention.
• compositions comprise, in addition to the active agent, a physiologically acceptable and preferably cosmetically acceptable medium, that is to say which does not cause unacceptable sensations of discomfort for the user such as redness, tightness or tingling.
• compositions according to the invention may contain as adjuvant at least one compound chosen from:
• oils which may be chosen in particular from silicone oils, linear or cyclic, volatile or non-volatile;
• waxes such as ozokerite, polyethylene wax, beeswax or carnauba wax;
• surfactants preferably emulsifiers, whether they are nonionic, anionic, cationic or amphoteric; coenzos, such as linear fatty alcohols;
• humectants such as polyols such as glycerine
• compositions are especially intended to promote the protection of the telosome and telomeres, in particular to prevent and / or fight against the effects of age on the skin.
• the invention aims at a cosmetic process for the care of human skin, intended to prevent and / or fight against the effects of age on the skin, in particular by promoting the protection of the telosome and the telomeres, including the topical application of a composition comprising non-sulfated linear galactans obtained from Eucheuma cottonii or an Eucheuma cottonii hydrolyzate comprising linear galactans according to the present invention.
• An example of a process for obtaining an active ingredient according to the invention comprises the implementation of the following steps:
• This thick, melting, white silicone emulsion has a pH of 6.8. In topical application on the skin, it has a fast penetration with a soft finish.
• Example 3 Use of an Active Ingredient According to the Invention in a Day Cream
• This creamy white emulsified gel has a pH of 6. In topical application, it has a velvety application with a soft finish and a powdery effect.
• This white, fluid emulsion has a pH of 6.8. It emulsifies easily with water, in the form of soft and fine foam, with a dry finish.
• Example 5 Use of an active ingredient according to the invention in an anti-wrinkle serum
• This white fluid gel emulsion has a pH of 5.5. As a topical application, it has a comfortable spread with a slightly friable cottony finish. It can be obtained by implementing the following steps:
• Phase E active principle according to the invention (example 1) 3%
• This white fluid emulsion has a pH of 6.8.
• the first goal of the study is to evaluate the expression of the two major telosome proteins responsible for protecting the telomeres POT1 ("Protection Of Telomeres” Protection of Telomeres) and TPP1 ("PTOl and TIN2-interacting protein” interacting protein with PTO1 and TTN2) in senescent human fibroblasts obtained as a result of moderate and repeated aggression with hydrogen peroxide (H 2 O 2 ) in comparison with normal human fibroblasts.
• the second aim of the study is to evaluate the ability of an active ingredient obtained from Eucheuma cottonii comprising linear galactans to limit the expression of PTO1 and TPP1 of normal human fibroblasts assaulted by the same treatment. hydrogen peroxide.
• normal human fibroblasts are inoculated in complete medium. The cells are then incubated at 37 ° C.
• Non-attacked controls normal fibroblasts are treated in complete medium for 2 hours, in the presence or absence of the active ingredient of Example 1 at 0.25% (V / V), and at the end of incubation. , the culture medium is removed and replaced with medium containing or not containing the active principle of Example 1 at 0.25% (V / V),
• Fibroblasts attacked and treated normal human fibroblasts are attacked and treated in complete culture medium with an H 2 O 2 solution for 2 hours in the presence or absence of the active ingredient according to Example 1 at 0.25% and 0.50%. At the end of the incubation, the culture medium is removed and replaced with medium containing or not containing the active principle of Example 1 at 0.25% and 0.50% (V / V).
• RNAs are reverse transcripts and the complementary DNAs obtained are analyzed by quantitative PCR (Quantitative Polymerase Chain Reaction).
• an active ingredient obtained from Eucheuma coffonii comprising linear galactans makes it possible to limit the alteration of the expressions of POT1 and TPP1, essential proteins of the telosome.
• Example 1 tested at 0.5%, the active principle of Example 1 makes it possible to limit by 100% the impairment of POT1 expression and by 89% the impairment of TPP1 expression.
• This second study aims to evaluate the ability of an active ingredient containing linear galactans obtained from Eucheuma cottonii to limit the acceleration of telomere erosion induced by repeated stress.
• the study was carried out by flow cytometry on pre-senescent human fibroblasts obtained following a moderate and repeated treatment with hydrogen peroxide (H2O2), according to the operating protocol described below. At 0, normal human fibroblasts are inoculated in complete medium. The cells are then incubated at 37 ° C.
• H2O2 hydrogen peroxide
• Non-attacked controls the normal fibroblasts are treated in complete medium for 2 hours, in the presence or absence of the active principle of Example 1 at 0.25% (V / V), and at the end of the incubation, the culture medium is removed and replaced with medium containing or not containing the active ingredient of Example 1 at 0.25% (V / V),
• Fibroblasts attacked and treated normal human fibroblasts are attacked and treated in complete culture medium with a solution of H2O2 for 2 hours in the presence or absence of the active ingredient according to Example 1 at 0.25% and 0.50% .
• the culture medium is removed and replaced with medium containing or not containing the active principle of Example 1 at 0.25% and 0.50% (V / V).
• the length of the telomeres is quantified using a dedicated kit.
• normal human fibroblasts are inoculated in complete medium. The cells are then incubated at 37 ° C.
• Non-attacked controls the normal fibroblasts are treated in complete medium for 2 hours, in the presence or absence of the active principle of Example 1 at 0.25% (V / V), and at the end of the incubation, the culture medium is removed and replaced with medium containing or not containing the active ingredient of Example 1 at 0.25% (V / V),
• Fibroblasts attacked and treated normal human fibroblasts are attacked and treated in complete culture medium with a solution of H2O2 for 2 hours in the presence or absence of the active ingredient according to Example 1 at 0.25% and 0.50% .
• the culture medium is removed and replaced with medium containing or not containing the active principle of Example 1 at 0.25% and 0.50% (V / V).
• the ⁇ -galactosidase activity is visualized using a dedicated kit. Visualization of the ⁇ -galactosidase activity is performed on a microscope coupled to an image analysis system. The activity of ⁇ -galactosidase is proportional to the dark blue coloring. Quantitative image analysis is also performed using software.
• This study aims to evaluate in vivo the ability of an active ingredient obtained from Eucheuma cottonii containing linear galactans, formulated at 3% emulsion, to promote the restructuring of the dermal matrix disorganized with age.
• the study was performed against placebo, at the level of the cheeks, on two groups of healthy volunteers of female sex: - a group treated with a placebo emulsion: 21 volunteers of average age 59 years
• Example 6 a group treated with the composition of Example 6 comprising the active ingredient of Example 1 in 3% emulsion: 24 volunteers of mean age 59 years old. Measurements of the acquisition of the fiber network at the level of the cheeks were made from sections, using a confocal microscope, equipped with 3 laser diodes. For the observation of the papillary dermis and the evaluation of the collagen fibers, the wavelength of 785 nm was chosen.
• the volunteers do not apply any cream or makeup on the face, and acquisitions are made by confocal microscopy on the cheeks.
• the placebo or emulsion containing the active ingredient of Example 1 at 3% is applied twice a day.
• telomere length can be used as an indicator of the replicative potential of cells and thus be associated with aging.
• the objective of the study is to evaluate in vivo the anti-wrinkle effect of an active ingredient obtained from Eucheuma cottonii comprising linear galactans formulated at 3% in emulsion against placebo at the level of the crow's feet by projection of fringes.
• Example 6 a group treated with the active ingredient of Example 1 in a 3% emulsion (composition of Example 6): 24 volunteers of mean age 59 years old.
• the acquisitions on the crow's feet were made using a fringe projection device dedicated to the 3D measurement of the cutaneous relief.
• This system includes a measurement sensor combining a bright fringe projector and a high resolution CCD camera connected to an acquisition software.
• o Sa arithmetic average of surface roughness
• a decrease in these various parameters is characteristic of an improvement in the relief of the studied surface and a decrease in wrinkles.
• Example 1 the placebo or the emulsion containing 3% of the active principle of Example 1 is applied twice a day.
• Example 1 formulated at 3% emulsion, compared with placebo, makes it possible to reduce the Sa parameter by 8.9%, the Sq parameter by 11.0% and the Sq parameter by 27.9%. negative volume parameter.

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