WO2008015343A2

WO2008015343A2 - Use of a yeast extract as active agent for increasing melanin synthesis in melanocytes

Info

Publication number: WO2008015343A2
Application number: PCT/FR2007/001335
Authority: WO
Inventors: Claude Dal Farra; Nouha Domloge; Dominique Peyronel
Original assignee: Societe D'extraction Des Principes Actifs Sa (Vincience)
Priority date: 2006-08-03
Filing date: 2007-08-02
Publication date: 2008-02-07
Also published as: WO2008015343A3

Abstract

The present invention relates to the use of a yeast extract in a cosmetic composition or for the preparation of a pharmaceutical composition, as active agent intended to promote increased melanin synthesis in the melanocytes of the epidermis or the hair bulb. The yeast extract is mainly constituted of compounds of protein and peptide nature and is obtained from yeast of the genus Saccharomyces or Kluyveromyces.

Description

USE OF A YEAST EXTRACT AS AN ACTIVE AGENT TO INCREASE THE SYNTHESIS OF MELANIN IN

MELANOCYTES

The present invention relates to the use of a yeast extract in a cosmetic composition; the extract or composition being intended to promote the increase of melanin synthesis in melanocytes of the epidemic or hair bulb, with a view to giving it a tanned appearance, or to prepare the skin for exposure to the sun. The present invention also relates to the use of a yeast extract for the preparation of a pharmaceutical composition; the extract or the composition being intended to promote the increase of melanin synthesis in the melanocytes of the epidermis or of the hair bulb for therapeutic purposes, for repigmenting depigmented skin, for example in the case of vitiligo, or for pigmenting the hairs or hair, especially in the treatment and prevention of canities. The yeast extract is mainly composed of protein and peptide compounds and is obtained from yeasts selected from the genera Saccharomyces or Kluyveromyces.

The color of human hair and skin is a function of various factors including seasons of the year, race, sex and age. It is mainly determined by the concentration of melanin produced by melanocytes. Melanocytes are specialized cells that, via particular organelles, melanosomes, synthesize melanin. The synthesis of melanin or melanogenesis is a complex process whose precise mechanisms are not yet elucidated and which schematically involves the following steps:

Tyrosine Dopa ^ * ^"* ^Dopaquinone" Dopachrome * ^"Melanin in the epidermis, the melanocyte is involved in the epidermal melanin unit that comprises a melanocyte surrounded by about 40 neighboring keratinocytes. Gradually, as melanin is synthesized in melanosomes move from the perinuclear region to the end of the melanocyte dendrites.Phagocytosis, the end of the dendrites is captured by keratinocytes, degraded membranes and redistributed melanosomes in the keratinocytes, where they will provide protection The natural desquamation of the cells The production of melanin and its transport are regulated by various factors such as, for example, UV radiation, hormones or chemicals. Thus, an increase in exposure to UV radiation causes the synthesis of pigments and browning of the skin. The pigmentation of hair and hair requires the presence of melanocytes in the bulb of the hair follicle. The hair follicle is a tabular invagination of the epidemic that penetrates to the deep layers of the dermis. The melanocytes in the bulb of the hair follicle are in an active state, that is, they synthesize melanin. These pigments are transmitted to the keratinocytes intended to form the hair shaft, which will lead to the growth of a pigmented hair or hair.

It is accepted that canitia (natural whitening of hair) is associated with a decrease of melanin in the hair shaft.

Nowadays, it is important to look good and tanned skin is always a sign of good health. However, natural tanning is not always without risk insofar as it requires prolonged exposures to UV radiation, which are likely in return to induce an alteration of skin cells, especially in the case of individuals of clear phototypes. The skin ages prematurely, becomes dry and is characterized by many wrinkles and age spots. It is therefore desirable to find an alternative to natural tanning that is compatible with the health requirements of individuals. On the other hand, we know that in most populations the maintenance of a constant coloring of the hair, sign of youth, is a strong aspiration.

The search for compounds that can promote the synthesis of melanin in the skin and hair is a concern of dermatology and cosmetics. These new compounds are particularly useful as an alternative to sun exposure, to prepare the skin and protect it from the sun's rays, to obtain a tan more intense after exposure to the sun, to prolong the natural pigmentation of the skin after exposure to the sun. sun or to prevent and / or limit and / or stop the development of canities and even maintain the natural pigmentation of hair and / or gray or white hairs. By natural pigmentation of the skin is meant the coloring of the skin or hair determined by the melanin concentration.

In this respect, many solutions have been proposed by adding exogenous dyes. By exogenous dye, a dye, or a chemical substance, is meant capable of reacting with the superficial layers of the epidermis to cause the appearance of a pigmentation of the skin completely independent of the presence of melanin. One of the best known exogenous dyes is dihydroxyacetone, or DHA. However, only the stimulation of the pigmentation of the skin and / or the hair by the natural way allows a real protection vis-à-vis the UV radiations and remains the ideal way of stimulation of the pigmentation. Thus, it has been proposed in the prior art the preparation and use of activators of melanin biosynthesis (FR828097, FR 2831438, FR 2845285) involving hormones (alpha MSH or its derivatives WO2006037188) or prostaglandins ( WO9511003).

On the other hand, there are pathologies related to pigmentation such as vitiligo which is an autoimmune disease characterized by the appearance on the skin of white patches related to a lack of pigmentation or pityriasis versicolor, a superficial mycosis. causing the appearance of light spots that may appear immediately or after exposure to the sun.

The main object of the present invention is to provide a new active ingredient that significantly promotes the synthesis of melanin in the melanocytes of the epidermis and the hair bulb. The inventors have indeed demonstrated a biological activity and, more particularly dermatological and cosmetic, a yeast extract. The use of a yeast extract in the field of cosmetics has already been described in numerous patents, in particular in skincare, as described in patent FR 2 826 576. However, to the knowledge of the Applicant, the use of yeast extracts to promote the increase of melanin synthesis in the melanocytes of the epidermis or hair bulb, has not yet been exposed.

By pro-pigmenting active agent is meant a compound capable of promoting the increase of melanin synthesis in the melanocytes of the epidermis or hair bulb.

By hair and hair is meant all the hairy appendages and in particular also the eyelashes and the eyebrows.

The compositions according to the invention may be applied by any appropriate route, in particular oral, parenteral or external topical, and their formulation will be adapted by those skilled in the art, in particular for cosmetic or dermatological compositions. Advantageously, the compositions according to the invention are intended for topical administration. They contain a physiologically acceptable medium, in particular a cosmetologically or pharmaceutically acceptable medium, in particular a dermatologically acceptable medium. Thus, the subject of the invention is primarily the use in a cosmetic composition of a yeast extract, consisting mainly of protein and peptide compounds; the extract or the composition being intended to promote the increase of melanin synthesis in the melanocytes of the epidemic or hair bulb. The term "extract" refers to any isolated substance obtained from the raw material that yeasts constitute. The active agent can be defined as being a molecule or a set of molecules capable of modifying or modulating the functioning of a biological system. According to an advantageous embodiment of the invention, the extract is a yeast extract belonging to the genera Saccharomyces or Kluyveromyces; preferably, the extract according to the invention is prepared from yeast of the species Saccharomyces cereviseae, Kluyveromyces lactis and / or Kluyveromyces marxianus.

Of course, the extract can be prepared from yeast of at least one of the many varieties and species belonging to the genera Saccharomyces or Kluyveromyces.

According to an embodiment of the presently preferred invention, the yeast extract is an aqueous extract. By aqueous extract is meant any combination of compounds soluble in water or in any solvent consisting wholly or partially of water. The extracts according to the invention are in particular purified aqueous extracts. In particular, aqueous solvents may be mentioned. By aqueous solvent is meant any solvent consisting wholly or partially of water. We can mention water itself, hydroalcoholic solvents in all proportions or solvents consisting of water and a compound such as propylene glycol or butylene glycol in all proportions. This extract can be obtained by dissolution in water, alcohol or ether, then by concentration of this solution involving evaporation or distillation.

According to an advantageous embodiment of the invention, the yeast extract is an extract composed for the most part of compounds of peptide nature.

By compounds of peptide nature is meant all compounds of protein nature, such as protein fragments, peptides but also free amino acids. The peptides, amino acids and protein fragments are determined according to conventional techniques, which are well known to those skilled in the art.

On the other hand, according to another characteristic, the yeast extract contains compounds of low molecular weight. According to another advantageous embodiment of the invention, the yeast extract contains a quantity of compounds of peptide nature representing between 30 and 70% of the total weight of the solids, more particularly this amount is between 40 and 50% of the total weight of the solids.

Any method of extraction or purification known to those skilled in the art can be used to prepare the extract according to the invention.

Thus, in a first step, the yeasts are cultured in a conventional manner in a medium adapted to their development, preferably in the presence of lactose. They are harvested by centrifugation and then suspended in a buffer solution, preferably a phosphate buffer. In a second step, these cells are exploded using a French press or using a ball mill, the majority of the insoluble membrane components being removed by centrifugation or by filtration. A high protein filtrate can then be collected and redissolved. A fraction, rich in peptide-like compounds, can then be isolated by precipitation in an alcoholic medium or with a saline solution. The soluble components and the nucleic acids are thus discarded. According to a variant of the process for obtaining the extract according to the invention, a dialysis step and a hydrolysis step by a cocktail of proteases can be added in order to obtain a fraction rich in peptide-like compounds. An additional purification step by a chromatographic type method can be envisaged.

According to another embodiment of the yeast extract according to the invention, the hydrolysis step may be carried out directly on the yeasts harvested after centrifugation or on the fractions obtained after cell bursting. Filtration and sterilization steps are subsequently performed. Thus, according to an advantageous embodiment of the invention, the yeast extract contains a quantity of protein and peptide compounds representing approximately between 30% and 70% of the total weight of the dry extract, more particularly this amount is between 40% and 50% of the total weight of the dry extract. The invention has the second object the use of an extract according to the invention for the preparation of a pharmaceutical composition; the extract or the composition being intended to promote the increase of melanin synthesis in the melanocytes of the epidermis or of the hair bulb in order to attenuate the pigmentary disorders associated with hypopigmentation of the skin, in particular vitiligo, or in order to pigment the hair or hair, especially in the treatment and prevention of canities. The invention also relates to the use in a cosmetic composition, or for the preparation of a pharmaceutical composition, of a yeast extract; the extract or the composition being intended to promote the pigmentation of the hairs and / or the hair.

The invention also relates to the use in a cosmetic composition, or for the preparation of a pharmaceutical composition, of a yeast extract; the extract or the composition being intended to prepare the skin for exposure to the sun.

The invention also relates to the use in a cosmetic composition, or for the preparation of a pharmaceutical composition, of a yeast extract; the extract or the composition being intended, by increasing melanin synthesis, to protect the skin from sunlight.

According to another form of the invention, the use of a yeast extract, or the composition containing it, is intended to protect the skin from environmental stresses.

According to an advantageous embodiment of the invention, the yeast extract is first solubilized in one or more cosmetically or pharmaceutically acceptable solvents such as water, glycerol, propylene glycol, butylene glycol, ethoxylated or propoxylated diglycols, ethanol, propanol or isopropanol.

According to yet another advantageous embodiment of the invention, the yeast extract is first solubilized in a cosmetic or pharmaceutical vector such as liposomes or adsorbed on powdery organic polymers, mineral supports such as talcs and bentonites, and more generally solubilized in, or attached to, any cosmetically or pharmaceutically acceptable carrier.

To give an order of magnitude, in the cosmetic or dermatological compositions of the present invention, the yeast extract is used in an amount representing from 0.0001% to 20% of the total weight of the composition, and preferably in an amount representing from 0.05% to 5% of the total weight of the composition. Thus, the content of protein and peptide compounds in the compositions of the the present invention will then be from 0.00003% to 14% and preferably from 0.015% to 3.5% of the total weight of the composition, for yeast extracts containing between 30% and 70% of compounds of protein and peptide nature. Or, the content of protein and peptide compounds in the compositions of the present invention will be from 0.00004% to 10% and preferably from 0.02% to 2.5% of the total weight of the composition, for yeast extracts containing between 40 and 50% of compounds of protein and peptide nature.

In addition to the major advantage of the pigmentation of the skin in the sun protection, a pro-pigmenting compound according to the invention offers outlets in self-tanning products and products promoting tanning. For this, the invention relates to the use of yeast extract for the manufacture of products for artificial tanning and / or browning of the skin.

The invention also relates to the use of a yeast extract to improve the intensity and / or the homogeneity and / or the persistence of the coloration produced on the skin and / or the hair.

Preferably, the compositions according to the present invention will be in a form suitable for topical administration to the skin, and cover all cosmetic or dermatological forms. These compositions must therefore contain a cosmetically and / or dermatologically acceptable medium, that is to say compatible with the skin, hair or hair. These compositions may especially be in the form of creams, oil-in-water emulsions, water-in-oil or multiple emulsions, solutions, suspensions, gels, milks, lotions, sticks or powders, suitable for application on the skin, lips and / or hair.

These compositions comprise the excipients necessary for their formulation, such as solvents, thickeners, diluents, surfactants, antioxidants, dyes, preservatives, perfumes. Of course, those skilled in the art will take care to choose any additional compounds, active or non-active, and / or their quantity, so that the advantageous properties of the mixture are not, or not substantially, impaired by the addition considered. The composition that may be used according to the invention may in particular consist of a composition for hair care, and in particular a shampoo, a conditioner, a setting lotion, a treatment lotion, a cream or a styling gel, a restructuring lotion for hair, a mask, etc. The cosmetic composition according to the invention can be used in particular in treatments using an application that is followed or not followed by rinsing, or in the form of shampoo.

It can also be in the form of dye or mascara to be applied with a brush or a comb, in particular on eyelashes, eyebrows or hair.

It is of course obvious that the extracts according to the invention, mainly consisting of protein and peptide compounds, may be used as the only pro-pigmenting agent or in combination with at least one other active agent, in a cosmetic composition. or for the preparation of a dermatological and / or pharmaceutical composition.

Advantageously, the compositions that can be used according to the invention also contain at least one other agent that promotes the pigmentation of the skin, hair and / or hair. Such compounds include substrates of tyrosinase, such as tyrosine or L-DOPA ₅ prostaglandins, or compounds activators of the cAMP pathway such as derivatives of pro-opiomélanocortines, adenosine, or forkoline or its derivatives. It may also be mentioned plant extracts such as bitter orange (citrus aurantium) or chrysanthemum (Chrysanthemum morifolium), described in particular in patents FR 2845285 and EP 1014934. Such compounds are also found in the family of exogenous dyes of the superficial layers. of the epidermis, such as dihydroxyacetone (DHA), erythrulose, extracts of henna leaves, described in particular in patents EP 0742002, FR 2779958.

The invention relates to a cosmetic process for increasing the synthesis of melanin in melanocytes and promoting the natural pigmentation of the skin and the hair, which consists in applying the composition as defined above to the skin or the hair. The invention relates to a cosmetic treatment method for preventing or treating hair and hair whitening, comprising applying to the skin the composition as defined above.

Other advantages and characteristics of the invention will appear better on reading the examples given by way of illustration and not limitation.

Example 1 Preparation of Yeast Extracts

The active agent is obtained from a yeast extract of the species Saccharomyces cerevisiae, Kluyveromyces lactis or Kluyveromyces marxianus. Of course, the extract can be prepared from yeast of at least one of the many varieties and species belonging to the genera Saccharomyces or Kluyveromyces.

The yeasts are cultured in a medium adapted to their development and then centrifuged to recover a biomass. The biomass is then milled in a ball mill. Then the ground material is resuspended in water at a concentration of 100 grams per liter before enzymatic hydrolysis at 40-60 ° C for 6 hours. After hydrolysis, the product is centrifuged and filtered before sterilization.

A hydrolyzate is thus obtained containing a quantity of protein and peptide compounds representing approximately between 30 and 70% of the total weight of the solids, more particularly this amount is between 40 and 50% of the total weight of the solids content. .

Example 2 Ex Vivo Study of the Effect of the Yeast Extract According to Example 1 on the Synthesis of Melanin The purpose of this ex vivo study is to demonstrate the increase in melanization provided by the extract according to the invention.

Protocol: Human skin biopsies 6 mm in diameter are maintained in culture ex vivo in the presence of a specific medium (DMEM lg / L, HAMF 12, FCS and antibiotics) on inserts deposited in 6-well plates. The biopsies are cultured for 48 hours and receive or not 2 applications per day of yeast extract according to Example 1 at a concentration of 1%. The biopsies are then fixed in 9% formalin, NaCl (150 mM) for 10 hours and then included in the paraffin. 3 μm skin sections thick are then made and melanin is colored specifically by the Fontana-Masson technique.

Results: Skin sections that did not receive the application of extract according to Example 1 have a low intensity coloration. On the other hand, the skin sections having received the extract applications according to the invention exhibit a markedly increased color intensity. In addition, melanin is located in the basal layer, but also transported in the supra-basal layers.

Conclusions: 48 hours of treatment led to a strong increase of melanin synthesis by melanocytes and to stimulate the whole process of distribution of melanin in the epidermis.

Example 3 Preparation of compositions

These compositions were obtained by simple mixing of the various components. The quantities indicated are given as a percentage of weight.

1-oil-in-water emulsion

Oil phase: β Montanov 68 (Cetearyl Alcohol and Cetearyl Glucoside) 5.00% H Jojoba Oil 5.00%

Vaseline oil 5.00%

Isopropyl Palmitate 7.00%

Aqueous phase :

M Glycerin 5.00% B Allantoin 0.10%

"Extract according to example 1 1.00%

• Sepigel 305 (Polyacrylamide and C 13-14 Isoparaffm and Laureth-7) 0.30%

B Conservative 0.50%

B Perfume 0.50% u Water qs 100%

2- Gel Carbopol Ultrez 10 (2% solids) 25.00% ^β Triethanolamine 0.50% m Extract example 1 5.00% m Preservative 0.20% m EDTA (sequestering agent) 0.10% m Perfume 0.50% a Water qs 100%

3- Lotion

B Mono Propylene Glycol 1.00%

B Allantoin. 0.30% B Glycerin 1.00%

B Cetiol HE (PEG-7 Glyceryl Cocoate) 1.00%

B Extract according to example 1 0.5%

B Conservative 0.20%

B Perfume 0.50% B Water qs 100%

4-Oil-in-water self-tanning emulsion Oily phase:

B Glycerol mono / distearate mixture 2.00% B Stearyl alcohol 1.00%

B Palm oil stearic acid 1.50% B PoIy dimethylsiloxane 2.00%

B Poly dimethylsiloxane mixture alpha-omega 3.00% dihydroxyl / cyclopentadimethylsiloxane (14.7 / 85.3) B Lauroyl isopropyl sarconisate 15.00%

Aqueous phase :

B Dihydroxyacetone 5.00%

B Glycerin 5.00%

B Hexadecyl alcohol phosphate, potassium salt 1.00% B Polyacrylic acid 0.30%

B Triethanolamine qsp pH: 7

B dl-alpha-tocopheryl acetate 0.50%

B Extract according to example 1 1.00%

B Demineralized water qs 100.00% 5-Self-tanning emulsion water-daαs-oil

Oily phase:

B Polydimethyl / methyleetyl methylsiloxane oxyethylenated 2.00% B Phenyl trimethylsiloxy trisiloxane 3.00%

B Lauroyl isopropyl sarcosinate 10.00%

B Isohexadecane 5.00%

B Cyclopenta dimethylsiloxane 3.00%

Aqueous phase: B Dihydroxyacetone 2.00%

B Glycerin 5.00%

B Magnesium sulphate 0.70%

B Extract according to example 1 1.00%

B Demineralized water qs 100.00%

6- Self Tanning Lotion B Dihydroxyacetone 5.00%

"Freeze-dried extract of Eclipta prostata 0.40% m Ethanol 96% 13.00% B Ethoxydiglycol 7.00%

"PPG-1-PEG-9-Lauryl Glycol Ether 5.00%

B Parabens 0.25%

"Extract according to example 1 1.00%

B Demineralized water qs 100.00%

7- Self-tanning oil-in-water emulsion B Water 66.80%

B Ethoxydiglycol 5.00% B Xanthan gum 0.30% B Cetyl hydroxyethylcellulose 0.30% B Cetyl alcohol 2.00% B Stearyl alcohol 2.00% B Octyl palmitate 2.00% B Glucam P-20 distearate 2.00% m Steareth-20 1.00% "Dimethicone 1.00%" Polysorbate 60 1.00%

"Dimethicone Copolyol / IPDI Copolymer 1.00% B Glycerylstearate and PEG-100 Stearate 0.25%" Actiplex 335 Lipo OP 0.1% "Tocopheryl Acetate 0.1%" Water 10.975%

• Dihydroxyacetone 2.00% m Nipaguard MPA 0.60%

• Panthenol, 50% aqueous 0.40%

• Sorbic acid 0.05%

"Sodium Metabisulfite 0.025%" Extract according to Example 1 1.00%

Claims

1. Use of a yeast extract, chosen from the genera Kluyveromyces or Saccharomyces, consisting mainly of protein and peptide compounds, alone or in combination with at least one other active agent, in a cosmetic composition, the extract or the composition containing it being intended to promote the increase of melanin synthesis in the melanocytes of the epidermis or hair bulb and to improve the natural pigmentation of the skin.

2. Use of a yeast extract, chosen from the genera Kluyveromyces or Saccharomyces, mainly consisting of protein and peptide compounds, alone or in combination with at least one other active agent, for the preparation of a dermatological composition and / or or pharmaceutical, the extract or the composition containing it being intended to promote the increase of melanin synthesis in the melanocytes of the epidermis or hair bulb and to attenuate the pigmentary disorders associated with hypopigmentation of the skin.

3. Use according to claims 1 or 2, characterized in that the yeasts of the genus Kluyvomyce belong more particularly to the species Kluyveromyces lactis and / or Kluyveromyces marxianus and that the yeasts of the genus Saccharomyces belong more particularly to the species Saccharomyces cerevisiae.

4. Use according to any one of the preceding claims, characterized in that the yeast extract contains a quantity of compounds of protein and peptide nature, representing between 30% and 70% of the total weight of the dry matter and more particularly between 40% and 50% of the total weight of the dry matter.

5. Use according to any one of the preceding claims, characterized in that the composition is in a form suitable for topical application comprising a cosmetically or dermatologically acceptable medium.

6. Use according to any one of the preceding claims, characterized in that the extract is used in an amount representing from 0.0001% to 20% of the total weight of the composition, and preferably in an amount representing 0.05 % to 5% of the total weight of the composition.

7. Use according to any one of the preceding claims, characterized in that the composition further contains at least one exogenous dye of the superficial layers of the epidermis and / or at least one pro-pigmenting active agent different from said yeast extract, selected from tyrosinase substrates or prostaglandins or activating compounds of the cAMP pathway or pigment plant extracts.

8. Use according to any one of the preceding claims characterized in that the extract or the composition containing it is intended, by increasing melanin synthesis, to prepare the skin for exposure to the sun or to protect the skin radiation from the sun.

9. Use according to any one of the preceding claims characterized in that the extract or composition containing it is intended to protect the skin from external aggressions, in particular UV rays.

10. Use according to any one of the preceding claims characterized in that the extract or the composition containing it are intended for artificial tanning and / or browning of the skin.

11. Use according to any one of the preceding claims characterized in that the extract or the composition containing it are intended to improve the intensity and / or homogeneity and / or the persistence of the pigmentation of the skin and / or hair.

12. Use according to any one of the preceding claims characterized in that the yeast extract or the composition containing it are intended to reduce a pigmentary disorder of the skin, especially vitiligo.

3. Use according to any one of the preceding claims characterized in that the extract or the composition containing it are intended to prevent or treat the whitening of hair and / or hair.