JP2001513536A - Use of the purple discolored plant extract for cosmetics and drugs, especially dermatology - Google Patents

Abstract

  (57) [Summary] The present invention relates to the use of the purple discolored plant extract for cosmetics and pharmaceuticals, in particular dermatology. More specifically, the present invention relates to the use in the above two fields for the purpose of promoting keratinocyte differentiation, preventing the action of free radicals, and / or promoting the synthesis of glycosaminoglycan. For this purpose, a murasaki motomoto discolored plant extract is added to the ingredients to promote hydration of the skin, to promote softness and firmness, or to prevent aging of the skin, or to heal the skin, i.e. the skin with ichthyosis or psoriasis. . The present invention relates to the use of said extracts for the preparation of culture media for cells, in particular keratinocytes, to accelerate or improve their differentiation.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001] The present invention relates to a new use of the purple discolored plant extract in cosmetics and pharmaceuticals, in particular in dermatology. The invention also relates to the use of said extract as a medium for skin cells.

[0002] Murasaki Omoto is a plant belonging to the family Commeliaceae, and is also called Rhoeo discolor Hance, or Rhoeo spathacea Stearn, or Tradescantia spathacea. The place of origin of this plant is Central America, but it is grown in Brazil. It is also found in New Caledonia. The traditional use of this plant has been used in the treatment of oral bleeding, coughing, lung disorders and uterine bleeding and is described in the following literature. The Atlas of Medicinal Plants of Middle America, Julia F. Morton, Charle
s C. Thomas, editor, 1981 pp. 71-72 Also known for healing injuries.

[0003] Reports on similar uses can also be found in the following documents: Chinese herbs of Hong Kong, Hong Kong, 1985, pp. 188

[0004] According to the literature of B. Weigner, J. Ethnophamacol., 1982, 6, 67-84, the purple plant extract is used as an abortion drug in Haiti, but is considered to have no antiestrogenic effect. However, it is thought that it has an action to excite the rat uterus. In detail, it is cited in US Pat. No. 4,999,423 as a raw material of anthocyanin acylate for producing cosmetics, pharmaceuticals, and food coloring. The insecticidal activity of water extracts of leaves and stems is also cited in Lloydia, 1950, 13, 89-162.

[0005] The inventors have surprisingly discovered that the plant extract has several effects and believe that it may have potential use in both cosmetics and pharmaceuticals, particularly in dermatology. Although the action of these plant extracts has been demonstrated by the inventors,
Furthermore, they have an effect on keratinocyte differentiation, a free radical inhibitory effect, and a remarkable effect on glycosaminoglycan synthesis, hereinafter referred to as GAGs.

[0006] As described above, the present invention is based on the idea that the purple-colored plant extract has a specific action on keratinocyte differentiation, and is also useful for healing skin disorders related to keratinocyte differentiation inhibition. Can be used. In particular, this differentiation is due to increased cell aggregation to the epidermis, regulation of the conversion of keratinocytes to keratinocytes by nuclear loss and increased keratinization, an increase in the number of stratum corneum forming the stratum corneum, It appears as a phenomenon that contributes to strengthening the function and strengthening the water barrier that prevents excess water loss from the epidermis. Furthermore, although expressed in the form that regulates the process of keratin synthesis by keratin sites in the hair follicle, these proteins are also the main components of the hair shaft, and their properties are necessary to optimize the hair. In addition, the inventors have discovered that these extracts promote, accelerate, and even improve the differentiation of skin cells, particularly keratinocytes, when cultured in culture media.

Furthermore, the inventors have demonstrated that the action of the purple discolored plant extract acts as a scavenger for free radicals, and that the extract is a cosmetic or dermatological product having a free radical scavenging action It has also been demonstrated that it is of value for use in the manufacture of products, specifically for the purpose of inhibiting the oxidative effects that cause skin aging and the production of highly reactive radical species. In particular, these products are effective in repelling toxic OH radicals.

Further, the present inventors have found that the purple discolored plant extract has a useful value in that it has an effect of promoting GAG production by fibroblasts in dermis in cosmetics and pharmaceuticals, in particular, dermatology, in particular, human dermis. It has also been demonstrated. Therefore, experts say that a complex macromolecule composed of protein moieties in which proteoglycans partially composed of GA are grafted through covalent bonds, that is, a disaccharide known as glycosaminoglycan (GAG). It is well known that they are molecules, but this is described in ED Hay's book ("Cell Biology of Extrace
llular Matrix ", Plenum Press (New York), 1991), and a paper by JE Silvert (" Structure and Metabolism of Proteoglycans and Glycosaminoglycans ", J
Invest. Dermatol., 1982, 79, pages 31s-37s).

[0009] Thus, GAGs are present as one of the epidermal and extracellular matrix components. GAG
Was found to be related to the site of collagen fibers as introduced by JE Scott in a paper (J. Biol. Macromol., 1991, 13, page 157-161). These are described in JG Haggerty et al. (J. Invest. Dermato., 1992, 99, pages 374-380).
As was announced by the authors, it was also confirmed between keratinocytes in the epidermis. GAGs have also been found in the extracellular matrix of the dermal papilla of the hair follicle, but their amounts vary as a function of the hair cycle. Also, their amounts are highest during the tissue regeneration phase, which is the time of hair growth. Regarding hair follicle proteoglycans and GAGs, we have already introduced JR Couchman's paper and M. Ty
lor et al.'s paper ("Glycosaminoglycan synthesis by cultured human hair follicl
e dermal papilla cells: comparison with non-follicular dermal fibroblast
s ", Brit. J. Dermatol. 1992 (May) 126 (5) 479-84).

[0010] It should be noted that the well-known minoxidil, which has some effects on hair growth and new hair growth, has been shown to promote the biosynthesis of glycosaminoglycans (Y. Mori et. al., Ann. NY Acad. Sci., 1991 (Dec.
26) 642 473-5). Taken together, from the author's point of view and other clinical trials, it is clear that glycosaminoglycans are involved in the periodic growth of hair. It was also found that the disaccharide unit of GAG was formed from hexamine (D-glucosamine or D-galactosamine). Hexoamines are often sulfated alternately with uronic acids (D-glucuronic or L-iduronic). Chain GAGs include hyaluronic acid, chondroitin 4-sulfate and 6-sulfate, dermatan sulfate, to a lesser extent, heparin and heparin sulfate, which are common in the skin. In particular, it is well known that hyaluronic acid is an essential component for healthy hair and its maintenance. This acid alone is synthesized without binding to the central protein.

GAG strongly associates with water molecules and gels, so that the dermis and epidermis are moisturized. A properly moisturized skin is evidence of good appearance and good mechanical properties as well as satisfactory physiological and functional status. Many researchers have found that GAGs decrease with aging of the skin (eg, Smith et al. J. Invest. Dermatol).
., 1962, 39, pages 347-350, Fleischmajer et al., In Biochim Biophys. Ac
ta, 1972, 279, pages 265-275, or Longas et al., Carbohydr. Res., 198
7, 159, pages 127-136).

[0012] Thus, enhanced GAG synthesis may confer the quality and properties of a healthy young person to dry skin or skin with defective mechanical or water barrier properties such as skin aging. Or prevent or cure hair growth disorders and, secondly, restore the gloss and softness of the hair.

[0013] The main object of the present invention is to solve the new technical problems encountered when formulating new cosmetic and pharmaceutical compositions. In addition, since these compositions possess the property of promoting keratinocyte differentiation, they cure skin disorders that accompany keratinocyte differentiation such as psoriasis, protect the skin,
In particular, it is expected to provide a moisturizing effect by restoring, maintaining, and strengthening the water barrier function, and in particular, by preventing excessive loss of water from the epidermis. An advantage of applying the latter phenomenon in practice is that it cures not only ichthyosis-like erythroderma skin but also psoriatic skin. In addition, these compositions improve the quality of the hair and thus improve its appearance.

[0014] In addition, a main object of the present invention is to solve a new technical problem found when promoting, accelerating, and improving the differentiation of skin cells, specifically, keratinocyte differentiation in culture.

Another object of the present invention is to provide a cosmetic or pharmaceutical ingredient, in particular, a dermatological ingredient having a free radical-scavenging action. In addition, the main object of the present invention is to provide a cosmetic or pharmaceutical ingredient, specifically a dermatological ingredient, to solve new technical problems existing when promoting GAG synthesis. is there. Accordingly, the present invention relates to promoting the GAG synthesis by synthesizing a component as a cosmetic or pharmaceutical, specifically a dermatological component, using a purple discolored plant extract.

These components improve the firmness and softness of the skin to prevent wrinkles, and in particular provide a smooth skin due to a physiological tightening effect or improve the moisture of the skin. Another object of the present invention is to provide a solution for healing cells, particularly fibroblasts in culture, to promote GAG synthesis. More specifically, the present invention relates to the use of a purple-colored plant extract as an active ingredient of a cosmetic ingredient due to its substantial properties.

[0017] From a different point of view, the first view suggests that the purple-colored plant extract may provide or improve moisture to the skin, prevent aging of the skin, prevent or heal wrinkles, and even tighten and soften the skin. Use as cosmetic to improve. The inventor was able to combine, in addition to these various actions, the actions of the purple discolored plant extract on keratinocyte differentiation, free radical scavenging, and GAG synthesis.

More specifically, the cosmetic ingredient of the present invention promotes the differentiation of keratinocytes and has the effect of strengthening the skin barrier of the stratum corneum, so that the skin loses moisture, retains moisture, and has a skin appearance. It helps improve gloss, strengthens skin barriers to prevent the ingress of irritants or toxic substances from the outside, and also protects the skin and the entire body. According to the present invention, the free radical-scavenging action of these components appears as an effect of preventing skin aging. The effects of the cosmetic ingredients according to the present invention on the promotion of GAG synthesis have the effect of improving the moisture of the dermis and epidermis, preventing wrinkles or preventing internal progression of wrinkles, and improving the firmness and softness of the skin. Appears. The latter effect creates a smooth skin with physiological tensioning effects.

In another aspect, the present invention relates to the use of Murasaki Omoto for pharmaceutical use, in particular, for the formulation of ingredients for dermatological use. Therefore, as another feature, the present invention relates to the preparation of a discolored plant extract of Murasaki motomoto, specifically a dermatological component, so that healing or preventing disorders due to dysfunction of keratinocyte differentiation, in particular, irregular Or prevent excessive desquamation,
Healing of dry ichthyosis-like erythroderma skin or psoriatic skin, ameliorating epidermal barrier dysfunction, and / or aging of the skin, in particular preventing or curing aging due to the detrimental effects of free radicals, and / or The purpose is to heal an individual suffering from GAG synthesis deficiency by reversing the adverse effects of said deficiency, in particular by improving the firmness and softness of the skin, preventing the formation of wrinkles or reducing the internal progression of wrinkles.

Therefore, the preparation component, specifically the dermatological component of the present invention, improves the barrier effect of the skin when the keratinocyte differentiation, the preparation component, specifically the dermatological component is considered, As a result, in addition to preventing water loss, the fact that this action can prevent irregular or excessive desquamation proves to be particularly effective.

The preparation component, in particular the dermatological component, according to the invention acts on free radicals, thereby providing a solution for the aging of the skin, in particular for all oxidative phenomena. It is effective for all skin treatments in which it is desirable to improve the smoothness or moisture of the skin by the action of the preparation component of the present invention, specifically, the dermatological component on GAG synthesis, specifically, by tightening effect. It is proved that there is. For the two aspects of cosmetics and preparations, in particular the dermatological aspect, it is also possible to utilize the extracts of the whole plant. The leaf extract of the present plant can also be used effectively.

According to an advantageous embodiment, the extract of the purple discolored plant is extracted using a single solvent or a mixed solvent having a polarity parameter (p) in the range of 4 to 8. For a definition of the polarity parameter (p), see a translation from the German original by Valeire Cottrell, published in 1988 by John Wiley and Sons (Practical High
Performance Liquid Chromatography (Veronika R. Meyer)).

The extract can be efficiently obtained by immersing a plant, ie, a part of a plant, in a single solvent or a mixed solvent according to the above definition and filtering the mixture, and then, if necessary, evaporating the solvent or the mixed solvent. Obtainable. Examples of solvents or mixed solvents that are particularly useful for producing an effective extract according to the present invention are as follows. C1-C4 alcohols, specifically ethanol, methanol, isopropanol,
Or alcohol-soluble alcohols of these alcohols, ethers, specifically ethyl acetate or isopropyl acetate, C2-C6 ketones, specifically acetone, or C2-C6 glycols, specifically ethylene glycol, Mixture of solvents.

[0024] Various component forms may be preferred. A preferred form of the present invention, specifically a topical form applied to skin tissues such as skin and mucous membrane. There are no restrictions on the appropriate topical formulation, including emulsions, creams, emulsions, salves, gels, lotions, ovules, and even treatment make-up ingredients. It is well known.

As a cosmetic or preparation ingredient, in particular a dermatological ingredient, 0.0% by weight
Concentrations of the purple discolored plant extract in the range from 01 to 5%, preferably in the range from 0.01 to 2%, are also contemplated, but this concentration is expressed in terms of the weight of the dry extract relative to said component. .

According to two aspects of the present invention, a particularly useful variant for cosmetics and preparations, the component comprises at least one product selected from the group: -Amino acids, in particular resin, proline, threonine, serine, arginine,
And derivatives thereof-depigmenting substances, in particular α-hydroxy acids such as butyric acid, malic acid, glycolic acid, citric acid, tartaric acid, salicylic acid and natural extracts containing them-glycerol, glyceryl acetate, urea, trehalose, dextran Humectants such as-vitamins, in particular vitamins A, B, C, D, E, F and their derivatives or their esters, phosphate-ceramides, in particular plant or synthetic ceramides-natural ceramides Synthetic effectors-oligomers of procyanidol and also catechol tannins, gallic tannins and their derivatives, in particular their esters-substances known to promote collagen synthesis, in particular triterpenes and their derivatives, Specifically, their ester-collagen synthesis Substances known to progress, in particular triterpenes and their derivatives, in particular glycosyl derivatives or asiatic acid, madekasic acid, such as asiatecoside and madecassoside, and their esters or Centella asiatica Extracts-Algal extracts, specifically microalgae extracts such as Euglena or Chlorella-Curcuminoids, vitamin E derivatives, vitamin C derivatives, or nordihydrog
A substance with a trapping action, such as ariaretic acid.

Finally, according to a last aspect, the present invention also relates to a culture medium for culturing cells or tissues, in particular skin cells, in particular keratinocytes, wherein the medium is adapted to initiate differentiation. It promotes, accelerates and improves the differentiation of this species, which is characterized in that it occupies an effective amount of purple vegetative extract. The medium contains a purple discolored plant extract that weighs 0.0001 to 1% of the total weight of the medium. The invention also relates to the large-scale cultivation of skin cells, in particular keratinocytes and / or fibroblasts, or to the production of artificial skin, or to the production of a reconstructed skin model characterized in that it uses a medium as defined above.

Finally, the invention relates to the promotion, acceleration and improvement of skin cells, in particular keratinocytes, and / or the synthesis of GAGs, in particular in the context of large-scale cultivation of skin cells, in particular in the context of keratinocytes and / or fibroblasts. By promoting the GAG synthesis of cells, it is possible to create artificial skin or a reconstructed skin model consisting of dermis and epidermis, and to use a medium defined above.

Although the purpose, features and advantages of the present invention are well known to those skilled in the art, some examples of the preparation of the present invention can be seen from the following examples. In these examples, percentages are by weight unless otherwise indicated.

[0030]

EXAMPLE 1 Preparation of a methanol extract of all plants 1 kg of all plants is ground for 24 hours and then immersed in 10 L of cold methanol. next,
Extract under reflux for 30 minutes. The mixture is filtered through paper and the solvent is evaporated, yielding a dry extract called MD 44/94. According to a variant of this process, the ground plants are treated with hexane and then immersed in methanol according to conventional techniques to perform degreasing. A second methanol extraction test is performed using another whole milled plant according to the steps described in section 1 above. A dry extract called MD 82/95 is obtained.

Example 2 Preparation of a water-glycol extract of purple mussels A water-glycol mixture of 1,3-butylene glycol and water at a ratio of 1: 1 was used to adjust the weight ratio of the plant to the solvent to 1:20. To make an extract. This extract is 20 at 80 ° C
Obtained by heating the mixture for minutes. Although the yellow extract is recovered, the extract can be easily added to cosmetic formulations.

Example 3 Demonstration of Effect of Murasaki Moto on Keratinocyte Differentiation In this test, a skin specimen collected during breast plastic surgery of a 22-year-old woman was used. The procedure below is similar to that of Detmar et al. (Eur. J. Dermatl. 1994 4 558-562).

On day D = −2, 300,000 cells of keratinocytes isolated from the above specimens were inoculated in 12 wells of a multiwell culture plate per well into a standard culture medium for keratinocyte culture. I do. This standard medium is well known to the expert. This medium has as its components an M199 medium supplemented with 2 mM L-glutamine, 10% calf serum, cholera toxin, hydrocortisone, and epidermal growth factor EGF.

On days D = 1, D = 5, D = 8 and D = 11, the same medium was replaced for the 6 wells of the untreated control wells among the 12 wells, and the remaining wells consisted of the treated wells. For the 6 wells, the same medium as that described above is replaced, except that the purple discolored plant extract obtained in Embodiment 1 is dissolved at 2.5 μg / ml and subjected to sterile filtration with a 0.22 μm filter.

On D = 12 days, the cells are counted, and the cell tissue is fixed with glutaraldehyde and osmium tetroxide, and then embedded in an epoxy resin. The resin mass thus obtained is cut into ultrathin sections and sampled with a device known as a microtome.

Next, the sections of the six control cultures and the six sections of the cultures treated with the purple discolored plant extract were examined under a transmission electron microscope. The same test was carried out with a solution in which the concentration of the purple discolored plant extract was changed to 10 μg / ml instead of 2.5 μg / ml. It can be seen that the differentiation of cells treated with the purple discolored plant extract obtained in Example 1 according to the invention is significantly more active than in the control. It can be seen that when treated with 2.5 μg / ml, the cell layer is both high in total thickness and number. It can be seen from the uniformity of the cell thickness during differentiation that the stratification is good. The keratinocytes in the upper layer are also keratinized well, but in the case of immersion culture, they do not reach the final differentiation stage at all.

The essential properties for good differentiation are: -Flattening of upper cells-Thickening of keratin (upper layer)-Expression of desmosome-Development of keratohyalin granules (middle layer)-Alignment of keratin filaments-Generation of phagocytic phenomena When normal human keratinocytes were cultured at 2.5 to 10 μg / ml, it was found that differentiation was significantly improved.

Example 4 Demonstration of the effect of the purple discolored plant extract on suppression of free radicals a) Test method Linoleic acid was dissolved in a zwitterionic surfactant (CHAPS, Sigma) and the pH was adjusted to 0.0 at pH 7.4.
6 M phosphate buffer is slowly added to obtain a microemulsion. The linoleic acid microemulsion prepared as described above (regardless of test substance content) was exposed to gamma-rays emitted from a two-curie source of cesium-137 (ie, 20,000 rad) for 18 hours. Therefore, OH free radicals are generated by radiolysis of water. When linoleic acid is decomposed by OH radicals, pentane is produced, which evaporates into the closed flask space. The released pentane is analyzed by gas chromatography. Thus, it is possible to determine the level at which the generation of free radicals due to the presence of the purple discolored plant extract is suppressed by comparison with the tests performed on the control microemulsion.

B) Conclusions The results for various microemulsion concentrations of the extract obtained according to Example 1 are shown in Table 1.

[0040]

【table 1】

Therefore, from these results, it is clear that the extract of the present invention has an inhibitory effect on the generation of free radicals, specifically, the generation of OH radicals. When linoleic acid is also present in the stratum corneum and cell membrane of the epidermis, various biological factors exposed from the present invention, specifically skin lipid cements and cell membranes, have harmful free radical effects, specifically OH radicals. Proven protection from action. Thus, the use of the present invention ensures the integrity and functionality of these elements.

Embodiment 5 Demonstration of the effect of the purple discolored plant extract on promotion of GAG synthesis a) Test method Regarding the production and secretion of labeled glucosamine regardless of the presence or absence of the substances which are the subject of this invention The uptake of is studied using human cells. Using a culture of human dermal fibroblasts, 3H-glucosamine is added thereto regardless of the presence or absence of the test substance, and the culture is performed. Next, the protein site of the proteoglycan generated by hydrolyzing the suspension using pronase is cleaved. The produced and secreted GAG, which incorporates more or less labeled glucosamine, is precipitated with cetylpyridinium chloride (CPC). The radioactivity of the treated and control cultures is counted, the radioactivity being proportional to the amount of synthetic GAG. In order to see the response of the cells, these cells are simultaneously tested for PDGF at a maximum of 25 μg / ml. PDGF (platelet-derived growth factor) (see PDGF-BB, Sigma, 4306) is a substance known to promote GAG synthesis by dermal fibroblasts.

B) Test Method Culture of normal human fibroblasts (NHF) is started three weeks after transplantation of healthy skin obtained during plastic surgery. Add 2 mM L-glutamine to E199 medium (designated E199c), and further add 10% v / v calf serum (FCS, Gibco) to culture the cells. The culture dish is left in an incubator in a CO2 atmosphere (5%) maintained at 37 ° C. After subculture (passage) by trypsinization, use NHF after 6 passages (represented by P1 to P6). The test is performed using NHF679-labeled fibroblasts from a remodeled 68-year-old woman.

The product with the MD82 / 95 code is the ethanol extract of Murasaki Omoto obtained in embodiment 2. Make a stock solution with DMSO at a concentration of the dried extract of 25 mg / ml. Since this product dissolves at such a concentration, prepare a dry product (17.4 mg / ml) as a preparation after filtering through a 0.22 μm filter. From this stock solution, make two dilutions with concentrations of 6.9 mg / ml and 1.74 mg / ml.

10,000 NHFs were added to a microplate (Falco in 100 μl of E199c medium supplemented with 10% FCS).
n 96 wells) and assay each (GAG, protein and viability). NFH was cultured for 24 hours in a 5% CO2 atmosphere.

After 24 hours, the medium was either serum-free, containing the test product dissolved in DMSO or, in the case of “control” cultures, the same amount of DMSO, and 4 μCi of 3H-glucosamide (see TRK 398 Amersham). Replace with 100 μl of E199 C medium. After culturing at 37 ° C for 48 hours, the suspensions are taken out as a set, and transferred to an Eppendorf tube with a screw cap. 200 μl of a solution containing 0.02% sodium azide in PBS buffer containing 0.2 mg / ml pronase (Sigma P5147) is added to each tester. After leaving the test tube to stand at 37 ° C overnight, the test tube was immersed in boiling water for 5 minutes to inactivate pronase.

After cooling at room temperature, a 1: 1: 1 mixture of hyaluronic acid, chondroitin sulphate and dermatan sulphate, ie 8 mg / ml each of 40 μl of a solution containing 100 mg / ml CPC (Sigma C 9002) Perform co-precipitation with the solution. The obtained mixture is stirred and cultured at room temperature for 30 minutes. Centrifuge the precipitate, remove the supernatant, and
Collect the pellet with 400 μl of CPC solution at a concentration of ml. After stirring for 5 minutes, the mixture is transferred to a scintillation flask, and 10 ml of scintillation liquid is added to count the radioactivity.

At the same time, Roehm NW et al. (Journal of Immunological Methods 1991, 142, 257-2
The cytotoxicity of the test product will also be evaluated in the test using XTT published by 65). This test measures the viability of cells.

C— Results and Statistical Test The activity (A) of the product is expressed as a percentage in the following formula.

(Equation 1) GAGs produced in control NHF The amount of GAG produced is expressed in cpm, whereas the amount of cellular protein is expressed in μg. Results for treated NHF (n = 6) and control NHF (n = 6) are compared by Student's t-test. The test value is p = 0.05. In the case of a significant difference (p <0.05), it is represented by S, and when it is not significant, it is represented by NS.

D- Results d-1) Cytotoxicity The XTT test performed at a concentration of 1.7 to 17.4 μg / ml as described above demonstrates that the extract according to the invention is not toxic.

D-2) First Test A test is performed on the NHF679 fibroblast cell line (68-year-old female, orthopedic) of the second subculture P2. Tables 2 and 3 show the results obtained for the extract obtained in Embodiment 2 and the results of PDGF as a positive control.

[0052]

[Table 2]

[0053]

[Table 3]

D-3) Second test This test is to confirm whether MD82 / 95 has an active effect on GAG synthesis. As in the first test, the NHF679 strain (68-year-old female, orthopedic) was used, but it was performed in the third passage P3. Table 4 shows the results. Table 5 shows the results obtained with PDGF.

[Table 4]

[0055]

[Table 5]

D-4) PDFG (25 ng / ml) as a positive control for conclusion criteria was tested in two tests (+ 99% and + respectively).
123%) significantly promotes GAG synthesis. MD82 / 95 also significantly accelerates GAG synthesis in both studies, but at concentrations of 1.7 μg / ml or higher (
+ 23% and + 16%). The maximal effect is 17.3 μg / ml (+ 53% and + 42%), and at this concentration there is no cytotoxicity.

As a result, the methanol extract of Murasaki Omoto was specifically 1.7 to 17.4 μg /
In the ml concentration range, it is an activator of glycosaminoglycan by human dermal fibroblasts, and the maximum activity is seen at 17.4 μg / ml. Therefore, according to the present invention, the extract containing purple discolored plant extract is included in the component,
When applied to the skin, it has the effect of promoting GAG synthesis, and therefore has the effect of optimizing the extracellular matrix of the skin. Thus, the skin is tightened by rehydration and the communication between cells is improved.

Example 6 Moist Cream Butylene Glycol Extract of Murasaki Omoto Obtained According to Example 2 2 g Vitamin A Palmitate 0.05 g Glycerol 3 g Butyric acid 3 g Plant ceramides 0.2 g Perfume 100 g in total

The cream gives moisture to the skin three days after application, makes the wood finer, and further increases the gloss of the face.

Example 7 Anti-aging skin tightening cream Methanol extract of Murasaki Omoto according to Example 1 MD82 / 95 0.2 g Ascorbic acid phosphate 2 g Maleic acid 3 g Centella asiatica 0.5 g Excipient 100 g in total amount

When the present cream is applied to the face, the skin becomes firm and wrinkles and fine lines disappear.

Example 8 Treatment base cream Murasaki Omoto methanol extract MD44 / 94 according to Embodiment 1 0.1 g Vitamin E acetate 0.1 g Curcuma longa extract 0.2 g Nylon particles 2 g Pigment-containing liquid excipient 100 g in total

This foundation cream promotes the gloss of the skin while preventing the signs of aging.

──────────────────────────────────────────────────続 き Continued on the front page (72) Inventor Pierre, Potier France, Paris, Abnu, de, Bourtouille, 14 (72) Inventor Françoise, Pico France, Brussels, Rue, de, Dan Pierre, 50 ( 72) Inventor Jean-Pierre, Cosson Gif-sur-Yvette, Battyman, The, Residence, Siroff F-term (reference) 4B065 AA93X BB26 CA50 4C083 AA111 AA112 AC121 AC122 AC272 AC292 AC301 AC421 AC471 AC581 AC641 AC642 AC681 AD072 AD211 AD241 AD391 AD531 AD611 AD621 AD622 AD631 AD641 AD642 AD651 AD661 AD662 AD701 CC05 DD31 EE12 FF01 4C088 AB71 BA10 CA04 CA06 CA07 MA02 MA63 ZA89

Claims (15)

[Claims] 1. Use of a purple discolored plant extract as an active principle in cosmetic compositions. 2. The extract is used for moisturizing or improving skin moisturization, preventing skin aging, preventing or healing wrinkles, tightening skin and imparting flexibility, Use according to claim 1, for use as a cosmetic substance. 3. Use according to claim 1 or 2, wherein the extract is used for the purpose of: strengthening the skin barrier of the stratum corneum, thereby reducing skin water loss, Keratinocytes, which maintain the wettability of the skin, improve its appearance and luster, strengthen the skin barrier against invasion of irritating or toxic substances from the outside, and also generally protect the skin and body. Promoting differentiation, and / or preventing aging of the skin by preventing the detrimental effects of free radicals, and / or improving the synthesis of glycosaminoglycans (GAGs), Improves epidermal wetting, controls wrinkle formation or reduces wrinkle depth, improves skin firmness and softness, and lubricates skin with physiological tightening effects Be in either. 4. Use of the extract of the purple discolored plant for the preparation of a pharmaceutical composition, in particular a dermatological composition, for the following purposes: a disorder associated with a dysfunction of keratinocyte differentiation. Preventing or healing, especially preventing irregular or excessive desquamation, healing the skin of dry ichthyosis or psoriasis, improving the protective function of the epidermis, or treating epidermal barrier dysfunction; and / or Prevent or cure skin aging, especially aging associated with the deleterious effects of free radicals; and / or treat individuals suffering from glycosaminoglycan (GAG) synthesis deficiency to reduce the adverse effects of said deficiency. To correct, especially to improve the firmness and softness of the skin, to prevent the formation of wrinkles or to reduce the depth of wrinkles. 5. Use according to any one of the preceding claims, wherein the extract is a whole plant extract. 6. Use according to any one of the preceding claims, wherein the extract is an extract of the leaves of the plant. 7. The extract, wherein the plant or part of the plant is softened by immersing it in a solvent or a mixture of solvents having a polar parameter p ′ of 4 to 8, followed by filtration and, if necessary, Or obtained by evaporating a mixture of solvents.
Use according to any one of the preceding claims. 8. The use according to claim 7, wherein the solvent or mixture of solvents is selected from the group of solvents consisting of: C1-C4 alcohols, in particular ethanol, methanol, isopropanol, Or aqueous alcohol mixtures of these alcohols,-ethers, especially ethyl acetate or isopropyl acetate,-C2 to C6 ketones, especially acetone, or-C2 to C6 glycols, especially ethylene glycol, and the above solvents. Mixture. 9. The composition according to claim 1, wherein said extract is present in a cosmetic or pharmaceutical composition, in particular in a dermatological composition, in an amount of 0.001 to 5% by weight, preferably 0.1% by weight, based on the weight of said composition. 01-2
9. Use according to any of the preceding claims, wherein it is present in a concentration expressed as% by weight of the dry extract. 10. The method according to claim 1, wherein said cosmetic or pharmaceutical composition, in particular a dermatological composition, also comprises at least one product selected from the group consisting of: Uses described:-amino acids, especially resins, proline, threonine, serine, arginine and their derivatives;-exfoliants, especially alpha-hydroxy, such as butyric, malic, glycolic, citric, tartaric, salicylic acid. Acids and natural extracts containing them,-humectants such as glycerol, glyceryl acetate, urea, trehalose, dextran-vitamins, especially vitamins A, B, C, D, E, and F, and derivatives thereof, especially Their esters or phosphates, ceramides, especially ceramides of plant or synthetic origin, natural synthetic effectors of ceramides -Procyanidol oligomers and catechol tannins, gallic tannins and their derivatives, especially their esters,-substances known to promote collagen synthesis, especially triterpenes and their derivatives, especially glycosyl derivatives or asia Asian Acid or Madekasic Acid, such as Tecoside and Madecassoside,
And their esters or extracts of Centella asiatica;-algal extracts, especially extracts of microalgae such as Euglena or chlorella;-curcuminoids, vitamin E derivatives, vitamin C derivatives or nordihydrog.
A substance that has a radical scavenging action, such as ariaretic acid. 11. Use according to any one of the preceding claims, wherein the cosmetic or pharmaceutical composition, in particular a dermatological composition, is formulated for topical application. 12. A medium for culturing cells or tissues, especially for culturing skin cells, in particular keratinocytes, for promoting, accelerating and improving their differentiation, said medium for obtaining said differentiation. , A medium comprising an effective amount of a purple discolored plant extract. 13. The medium according to claim 12, comprising 0.0001 to 1% by weight, based on the total weight of the medium, of the purple discolored plant extract. 14. Use of the medium according to claim 12 or 13 for bulk culture of skin cells, in particular keratinocytes, for the production of artificial skin or for the formation of a reconstructed skin model. Features method. 15. Skin cells, in particular for promoting, accelerating and improving the differentiation of skin cells, in particular keratinocytes, and / or for promoting GAG synthesis, especially from keratinocytes and / or fibroblasts. A method for producing an artificial skin or forming a reconstructed skin model in the bulk culture of the present invention, wherein the medium according to claim 12 or 13 is used.