FR2851576A1 - Preparing agent for reducing alterations in the contours of the eye, useful e.g. for reducing dark rings under the eyes, by solubilizing yeast in basic medium then enzymatic hydrolysis - Google Patents

Abstract

Method for preparing an active agent (A) for controlling alterations of the contours of the eyes, especially rings or pronounced pigmentation in this area, comprises solubilizing yeast, preferably Saccharomyces cerevisiae, in aqueous basic medium at at least 200 g/l; hydrolysis with at least one enzyme; separation of solids and liquids by filtration, decantation or centrifugation, then sterile filtration. Independent claims are also included for the following: (1) (A) prepared by the new method; and (2) cosmetic composition containing (A). ACTIVITY : Dermatological. A composition containing 4% (A) in a carbopol gel was applied twice daily to the rings around one eye, while the same regions around the other eye were treated with a placebo. After 28 days, color was measured with a Chromametre CR300, by determining the parameter L (lightness). For the treated eye, L had increased from 57.83 to 58.06 (0.4% increase) but for the placebo it had decreased from 58.21 to 56.57 (a 2.8% reduction). MECHANISM OF ACTION : (A) stimulates formation of new blood vessels and restructuring of the capillary network, and also stimulates vascular endothelial growth factor, thus normalizing neovascularization.

Description

PROCESS FOR OBTAINING AN ACTIVE INGREDIENT FROM YEASTS FOR

  FIGHTING EYE CONTOUR ALTERATION

  The present invention covers a process for obtaining an active principle from yeasts, more particularly Saccharomyces Cerevisiae, making it possible to combat alterations in the contour of the eyes, that is to say essentially against dark circles and marked pigmentation in this particular area. .

  The invention also covers cosmetic products which incorporate such an active ingredient in the form of creams, ointment, lotions or any other galenical form, including in combination with other active agents having a synergistic effect or simply a complementary action.

  The eye area is a particularly sensitive and complex area, the aesthetic role of which is very important since it affects the look and transmits a great deal of information concerning the person. This area, however small, is nevertheless an immediate reflection of fatigue, mood and of course age.

  It is therefore an area to which women are very attentive.

  This area of the eye contour has a very thin skin thickness since it is of the order of half a millimeter. The connective tissue is mainly composed of collagen, elastin and glycosaminoglycans This area is very vascular to supply a large network of over thirty muscles, half of which is dedicated to opening / closing the eyes.

  Therefore, when the tissue of this highly vascularized area is found to be lacking in certain compounds promoting its elastic properties, there is a sagging of the skin.

  Defects in circulation at the level of the capillary vessels of this zone 5 cause a change in color by the stagnation of the blood, by a higher concentration of melanin, by a concentration in ferric pigments, which reinforces the defects which appear in parallel with the games. of induced light. The skin is often duller, these are dark circles.

  In addition, when certain muscles relax, in particular the muscle called 10 Orbital septum, a pocket is formed.

  There are of course make-up products which moisturize the skin and / or mask certain flaws or even compensate for a lack of radiance with light-reflecting products or lightening white pigments or products with depigmenting activity. .

  However, no more in-depth treatment is planned to act on the causes of these alterations.

  We also know that other factors are involved in the appearance of dark circles or the formation of puffiness, in particular fatigue, lack of sleep, stress and, as mentioned above, age. Pollution and the consumption of tobacco also favor such alterations.

  There is therefore an expectation for products which make it possible to effectively combat the effects induced by these factors.

  The present invention relates to a process for obtaining an active ingredient derived from yeasts which makes it possible to improve the appearance of the contour of the eye.

  Such an active agent makes it possible to reshape the capillary network and to act on the hyperpigmentation of the suborbital zone by having simultaneously an action of standardization of the color of this zone.

  The appended drawings show on the different results obtained. FIGS. 1A to 1E thus show views of the different developments of the capillary network.

  To this end, the process provides for the use of yeast extracts, more particularly Saccharomyces Cerevisiae.

  1 / Process for Obtaining the Active Principle According to the Invention The process consists of the succession of the following stages - solubilization of yeasts, more particularly Saccharomyces Cerevisiae, in a basic aqueous medium at a rate of at least 200 g / l, 10 - enzymatic hydrolysis (s), - separation of the solid / liquid parts by filtration, decantation or centrifugation, and - sterilizing filtration.

  2 / Chemical composition of the active ingredient obtained by the process 2-1 / Dry matter content The dry matter content is achieved by weighing 10 g of product placed in an oven at 105 C until a constant weight is obtained .

  In the present case, the amount of dry matter is from 20 to 200 g / l, more particularly 38 to 52 g / l.

  2-2 pH measurement The pH is measured by the potentiometric method, at room temperature.

  In the present case, the pH is between 4.0 and 9.0, more particularly in the range of 6.0 to 7.0.

  2-3 Determination of protein content The measurement of protein content is determined by the Lowry method.

  In the present case, the protein content is between 7.0 and 85 9 / I, more particularly between 14 and 22g / l.

  2-4 Determination of the total sugar content The reducing sugars are measured by the bubois method. In the present case, the content of reducing sugars is between 9.0 and 96 9 / l, more particularly between 17 and 25 g / l.

  2-5 Determination of the amount of vitamin PP Vitamin PP is dosed according to the SNELL method (Snell E. et ai, 1941, Biol, Chem, 139,675).

  in the present case, the content of vitamin PP in the active principle according to the present invention is between 500 and 4,000 ng / ml, more particularly 1000 ng / ml and 2,000 ng / ml 3 / Effects obtained by the use of the active ingredient 3-1 Effects on neovascularization Neovascularization consists in determining the formation of new vessels from the pre-existing network.

  In in vitro culture, this results in endothelial cells by the formation of continuous tubules from these cells, thanks to migrations under the influence of different angiogenic factors. These cells can thus form blood vessels to form a capillary structure with ramifications.

  The test protocol consists in visualizing these endothelial cells as well as the branches and tubular structures formed. To this end, an immunostaining is carried out using an anti-PECAM-1 antibody (flat / and endothelial cel / adhesion molecule) which is a specific marker for these endothelial cells.

  These endothelial cells are cultured in the presence of the active principle according to the invention dosed at 0.1%; 0.5% and 1.0%.

  The comparison is carried out with the same endothelial cell culture medium in the presence of vitamin PP. The visualization is carried out under the microscope coupled to an image system. The number of tubular structures and the number of branches are measured. Tubular structures Nbr of branches Length (pm) Variation (%) Control 1172 _ 99 1.5 + 0.75 Vitamin PP 2068 244 + 76 8.75 + 1.84 Principle active 0.1% 1,225 + 97 + 5 1.75 0.29 Active ingredient 0.5% 1,859 220 + 58 7.25 + 1.85 Active ingredient 1.0% 2,026 35 + 73 10.25 0.55 On notes in the results measured from microscope views such as those of FIGS. 1A to 1E that the active principle causes the formation of new vessels and promotes the restructuring of the capillary network.

  FIG. 1A shows a view of a control, FIG. 1B shows the neovascularization obtained after treatment with vitamin PP. FIGS. 1C to 1E show neovascularizations with respectively 0.1%, 0.5% and 1.0% of active principle according to the present invention.

  3-2 Effects on VEGF (Vascular Endothelium Growth Factor) The objective is to determine the propensity of the active ingredient to stimulate the vascular growth factor also called VEGF. This factor is one of the factors regulating neovascularization.

  To do this, keratinocytes seeded in a culture medium are treated with the active principle according to the present invention dosed at 0.5% /; 1.0% / and 2.0%, diluted in this culture medium.

  After incubation, the cell supernatant is recovered and the VEGF assay is carried out using a specific ELISA assay kit. The following results are observed: VEGF (pg / g) | VEGF synthesis (/%) Control 2.73 Active ingredient 0.5% 3.30 +21 + 14 Active ingredient 1.0% 4.07 +49 9 Active ingredient 2.0% 0 4.37 +60 + 19 The synthesis of VEGF is stimulated in a proportion of 60% compared to the control with 2.0% of active principle, which is particularly attractive.

  3-3 Effects on darkening of dark circles The study is carried out in vivo against placebo.

  The concealer effect is evaluated by comparing the color of the dark circles before and after treatment with the active ingredient for one month.

  The formulation is 4% active ingredient in a carbopol gel or emulsion.

  The areas concerned are chosen on volunteers, one ring treated with the formulation with active ingredient and the other treated with placebo.

  The color of the skin is measured beforehand using an apparatus sold under the name Chromameter, model CR300.

  The staining results are then analyzed 28 days after twice daily treatment.

  The measured parameter is L: the area from dark to pale, (clarity) ci / Variation of L with the active ingredient with carbopol gel: LPr is obtained with the formula (J28 - OJ) / OJ API is also obtained with the formula ( J28 - OJ) / OJ We obtain the value of LA = L \ Pr -, PI or 3.2%.

  b / Variation of L with the active principle in emulsion: These values are calculated in the same way as above.

  It is noted that there is a clarification of the area of the dark circles by the application of the active principle, whatever the galenical formulation.

  3-4 Effects on the uniformity of the complexion The study consists in comparing the color of the area of a dark circle and the color of the skin on the rest of the face, more particularly on the crow's feet area.

  The active ingredient is formulated in a carbopol gel and dosed at 4%.

  The test is carried out on volunteers for 28 days by twice daily application of this formulation on specific areas, against placebo.

  The clarity L is then measured. The results are given in the following table: OJ J28 Face area Dark circle treated | O / Face area | Circle treated Active ingredient 61.14 57.83 5.4 61.50 58.06 5.6 Placebo 61.14 58.21 4.8 61, 70 56.57 8.3 Difference - - 0.6 - - - 2.7 After 28 days of treatment, it can be seen that the difference between the two zones remains constant 5.4 for 5.6% when the zone is treated with the active principle while it degrades 4.8 to 8.3 % when the area is treated with placebo. The dark circle appears darker.

  The active principle according to the present invention reduces the color of dark circles. There is a standardization of color differences, dark circles appear less marked.

  For all of the above tests, any formulation such as a gel, an emulsion, a cream, an ointment or a lotion can be used as dosage formulation. This choice will depend on the practicality, the destination and the best commercial reception. Such a choice falls within the competence of the final formulator.

  The active ingredient according to the present invention can be combined with any other active ingredient to obtain synergistic efficiencies.

Claims (8)

  1. Method for obtaining an active principle which makes it possible to combat alterations in the area around the eyes, in particular dark circles and marked pigmentation in this area, characterized in that it comprises the following stages: - solubilization of yeasts, more particularly Saccharomyces Cerevisiae in a basic aqueous medium at a rate of at least 200 g / t, - enzymatic hydrolysis (s), - separation of the solid / liquid parts, by filtration, decantation or centrifugation, and - sterilizing filtration.   2. Active principle from yeasts, more particularly Saccharomyces Cerevisiae making it possible to combat alterations in the eye area, obtained by implementing the method according to claim 1, characterized in that it comprises: - dry matter content between 20 and 200 g / l, 15 - the pH is between 4.0 and 9.0 - the protein content is between 7.0 and 85 g / I - the vitamin PP content of the active ingredient is included between 500 ng / l and 4,000 ng / l.   3. Active principle from yeasts according to claim 1, characterized in that it comprises: - dry matter content of between 38 to 52 g / I - the pH is between 6.0 and 7.0 - the protein content is between 14 and 22 g / l - the vitamin PP content of the active principle is between 1000 ng / l and 25 2000 g / l.   4. Cosmetic composition containing an active principle according to one of claims 2 or 3, obtained from yeasts in particular Saccharomyces Cerevisiae, according to the method of claim 1, characterized in that the active principle, associated with a suitable dosage form as a gel, an emulsion, a cream, an ointment or a lotion, promotes neovascularization by leading to the formation of new vessels and to the restructuring of the capillary network.   5. Cosmetic composition containing an active principle according to one of claims 2 or 3, obtained from yeasts in particular Saccharomyces Cerevisiae, according to the process of claim 1, characterized in that the active principle, associated with a suitable dosage form such that a gel, an emulsion, a cream, an ointment or a lotion, stimulates the vascular growth factor also called VEGF and regulates the neovascularization.   6. Cosmetic composition containing an active principle according to one of claims 2 or 3, obtained from yeasts in particular Saccharomyces 15 Cerevisiae, according to the method of claim 1, characterized in that the active principle, associated with a suitable dosage form such that a gel, an emulsion, a cream, an ointment or a lotion, stimulates the vascular growth factor also called VEGF and regulates the neovascularization.   7. Cosmetic composition containing an active principle according to one of claims 2 or 3, obtained from yeasts in particular Saccharomyces Cerevisiae, according to the method of claim 1, characterized in that the active principle, associated with a suitable dosage form such that a gel, an emulsion, a cream, an ointment or a lotion, leads to the lightening of dark circles.   8. Cosmetic composition containing an active principle according to one of claims 2 or 3, obtained from yeasts in particular Saccharomyces Cerevisiae, according to the method of claim 1, characterized in that the active principle, associated with a suitable dosage form such that a gel, an emulsion, a cream, an ointment or a lotion, leads to the standardization of the complexion of the dark circles area with the rest of the face.