COMPOSITION COMPRISING AT LEAST ONE GLYCOSYLATED FLAVONOID AND ITS USE IN COSMETICS OR DERMATOLOGY

Abstract

The present invention relates to a cosmetic and/or dermatological composition comprising as an active ingredient at least one glycosylated flavonoid, characterized in that it further comprises, as an active ingredient, a compound capable of limiting or preventing the penetration into the keratinocytes of a polycyclic aromatic hydrocarbon such as benzo[a]pyrene, cyclopenta[c,d]pyrene, dibenzo[a,h]anthracene or dibenzo[a,l]pyrene and/or a compound characterized by its ability to absorb blue light.

Claims

1. A cosmetic and/or dermatological composition comprising as an active ingredient at least one glycosylated flavonoid, and further comprising, as an active ingredient, a compound capable of limiting or preventing the penetration into the keratinocytes of a polycyclic aromatic hydrocarbon such as benzo[a]pyrene, cyclopenta[c,d]pyrene, dibenzo[a,h]anthracene or dibenzo[a,l]pyrene and/or a compound characterized by its ability to absorb blue light.

2. The cosmetic and/or dermatological composition according to claim 1, characterized in that said glycosylated flavonoid is a glycosylated flavonol.

3. The cosmetic and/or dermatological composition according to claim 1, characterized in that said glycosylated flavonoid is rhamnosylated or glycosylated, preferably rhamnosylated.

4. The cosmetic and/or dermatological composition according to claim 3, characterized in that said rhamnosylated flavonoid is chosen among myricitrin, quercitrin, kaempferitrin, azalein, icariin or a mixture of at least two of these compounds.

5. The cosmetic and/or dermatological composition according to claim 4, characterized in that said rhamnosylated flavonoid is myricitrin.

6. The cosmetic and/or dermatological composition according to claim 1, characterized in that said compound capable of limiting or preventing the penetration into the keratinocytes of a polycyclic aromatic hydrocarbon such as benzo[a]pyrene, cyclopenta[c,d]pyrene, dibenzo[a,h]anthracene or dibenzo[a,l]pyrene is chosen among polysaccharides, cyclic polysaccharides, such as cyclodextrin, emulsifiers, lyophilized hyaluronic acid.

7. The cosmetic and/or dermatological composition according to claim 1, characterized in that said compound capable of absorbing blue light is chosen among carotenoids or melanoidins.

8. The cosmetic and/or dermatological composition according to claim 1, characterized in that it is formulated for topical application.

9. A glycosylated flavonoid compound, preferably a rhamnosylated flavonol, for use in the treatment of non-allergic pathologies that trigger the production of IL31, in particular pathologies affecting the skin and chosen among pruritus, atopic dermatitis, eczema, psoriasis and nodular prurigo.

10. A glycosylated flavonoid compound, preferably a rhamnosylated flavonol according to claim 9, chosen among myricitrin, quercitrin, kaempferitrin, azalein, icariin or a mixture of at least two of these compounds.

Description

DESCRIPTION OF THE FIGURES

[0064] FIG. 1 presents a cell viability assay that measures the percentage of viable cells of human keratinocytes placed in the presence of myricitrin at different doses;

[0065] FIG. 2 presents the effect of benzo[a]pyrene, alone or in combination with blue light, on the production of IL-31 by human keratinocytes. Exposure to blue light is expressed in Joules per square centimeter (J/cm.sup.2.) Exposure to benzo[a]pyrene is expressed in micromoles (μμM);

[0066] FIG. 3 presents the effect, in pg/ml, of myricitrin on the release of IL-31 in keratinocytes that have been previously exposed to benzo[a]pyrene and blue light;

[0067] FIG. 4 presents the concentrations of IL31, standardized by the amount of total cellular proteins, illustrating the effects of astaxanthin alone compared to the effects of myricitrin alone, after exposure of keratinocytes to blue light and to benzo[a]pyrene;

[0068] FIG. 5 presents the inhibition value of the release of IL31 for two concentrations of myricitrin and astaxanthin.

EXAMPLES

Example 1: Culture of Human Keratinocytes

[0069] The cells used in this study are primary cultures of normal human keratinocytes (NHKs) extracted after skin surgery of a 30-year-old donor.

[0070] They were cultured in a complete KSFM environment: Keratinocyte-SFM (17005-31, Gibco) with L-Glutamine, supplemented with a recombinant human epidermal growth factor (EGFhr, 10450-013, Gibco), extract of bovine pituitary gland (BPE, 13028-014, Gibco) and 1% of streptomycin penicillin (15070-063, Gibco) at 37° C. in 5% CO.sub.2 atmosphere up until 80% confluence.

Example 2: Keratinocyte Viability Assay

[0071] On D0, normal human keratinocytes are incubated in 96-well plates in a culture medium. The cells are left to stabilize for 24 hours at 37° C. in 5% CO.sub.2 atmosphere, as described in Example 1.

[0072] On D1, the cells are treated with myricitrin dissolved in the culture medium at different concentrations.

[0073] The culture is kept for 24 hours at 37° C. and 5% CO.sub.2.

[0074] The NHK viability was assessed using a Cell Proliferation Kit II (XTT) (11465015001, Roche) in accordance with the manufacturer's instructions.

[0075] The XTT system, a colorimetric method, is a test used to quantify mitochondrial activity. This method, which is simple, accurate and allows for reproducible results, can be used as a viability assay. This assay is based on the breakdown of yellow XTT tetrazolium salt into orange formazan by the “succinate-tetrazolium reductase” system present in the cells' mitochondrial respiratory chain. Conversion thus only takes place in metabolically active cells, which means living cells.

[0076] The formazan derivate is measured by spectrophotometry (at 450 nm with 650 nm of reference.) For each condition, optical density data averages (OD, absorbance) are calculated.

[0077] The viability of the treated cells is expressed as a percentage of untreated control batch: a treatment that reduces the viability of the cells to below the comparative limit of 80% of mitochondrial activity in the untreated control batch, is considered cytotoxic for the cells. Inversely, an increase in the data is a sign of mitochondrial activity and possibly even a sign of cell proliferation.

[0078] Keratinocytes cultured according to Example 1 are exposed to a range of myricitrin of from 0.21 to 50 μμM. The Control batch represents the culture medium with no admixture. A 10% control+DMSO represents a toxic medium, indicating cell toxicity. Experience shows that myricitrin has no toxic effect on keratinocytes.

Example 3: Use of a Flavonoid such as a Rhamnosylated Flavonol on Cultured Keratinocytes

[0079] The keratinocytes cultured according to Example 1 are exposed either to benzo[a]pyrene (B[a]P) at 20 μμM, or to blue light (450 nm) at 40 J/cm.sup.2 or to the combination of both at the same doses. A myricitrin treatment with amounts ranging from 10 μμM to 50 μμM is applied to cells that have previously received the B[a]P and blue light combination.

[0080] After 24 hours' treatment, an assay of the interleukin 31 (IL-31) released by the cells is performed, via an immunoassay kit (ELISA.) The IL-31 assay was performed with the human IL-31 ELISA MAX™ Deluxe Set (445704, Biolegend.)

[0081] The combination of benzo[a]pyrene and blue light induces a +111% increase in IL-31 by the keratinocytes. The experiment shows that adding myricitrin as of 20 μM induces a −100% decrease in the production of IL-31 on cells pre-treated with 20 μM of B[a]P and 40 J/cm.sup.2 of blue light.

Example 4: Use of a Flavonoid such as Rhamnosylated Flavonol in Patients

[0082] In a group of volunteers working in urban areas [Caucasian and Asian women aged 20 to 60] and presenting with problems of cutaneous discomfort on the face (itching, pruritus, etc.), the cream in Example 5 is applied on one side of the face and a placebo cream with no myricitrin is applied on the other side of the face.

[0083] From the time of application and over eight days of morning and evening use, the volunteers record their organoleptic experience—freshness, gentleness and ease of application of the products—along with their physiological sensations such as calmness and relief from itching, on a questionnaire. Statistical analyses are performed at the end of the assay to demonstrate the efficiency of the product containing myricitrin.

Example 5: Calming and Softening Cream for Urban Skin or Skin Prone to Itching

[0084] A—Aqueous Phase

[0085] Glycerin 2.0%, Hexylene Glycol 3.0%, Xanthan Gum 0.5%, Myricitrin 0.5%, Preservatives qs (sufficient quantity), Carbomer 0.35%, NaOH 0.35%, Water qsp. 100%

[0086] B—Fatty Phase

[0087] Squalane 15%, Cetyl Alcohol 2%, Arachidyl Alcohol/Behenyl Alcohol/Arachidyl Glucoside 1%, Glycerol Stearate 5%, Preservative and fragrance qs

Example 6: Soothing and Moisturizing Cream for Urban Skin Prone to Atopic Dermatitis

[0088] A—Fatty Phase

[0089] Ceteareth-2 3.5%, Ceteareth-21 between 2 and 4%, Wheat Germ Oil 3%, Cyclomethicone 7%, Octyl Palmitate 8%

[0090] B—Aqueous Phase

[0091] Glycerin 7.0%, Hexylene Glycol 3.0%, Preservatives qs, Water qsp. 100%

[0092] C—Ingredients Added to the Emulsion, at a Temperature Below 40° C.

[0093] Sodium Hyaluronate 0.1%, Myricitrin 1.0%, Water 5%, Tocopherol 0.05%, Vitamin A Palmitate 0.1%, Phospholipids 0.5%, Ceramides 3 0.1%, Polyacrylamide & C14-13 Isoparaffin & Laureth-7 between 2 to 3.5%

Example 7: Cream and Milk for Skin Exposed to Blue Light or with Dark Spots

[0094] A—Fatty Phase

[0095] Glycerol Monostearate 2%, PEG-100 Stearate 3%, C12-C15 Alkyl Benzoate 10%, Dimethicone 5%, Tocopherol Acetate 1%, Octyl-Triazone (Uvinul T150) 1.5%, Butyl Methoxy Dibenzoyl Methane (Eusolex 9020) 2.0%, Ceostearyl Alcohol 1%

[0096] B—Aqueous Phase

[0097] Water qsp. 100%, Quercitrin 0.5%, Preservatives 0.6%, Glycerin 7%, Hexylene Glycol 3.0%, Carbomer 0.5%, Tetra Sodium EDTA 0.2%, Sodium Hyaluronate 0.1%, NaOH 0.5%, Preservative+fragrance qs.

[0098] C—Ingredients Added to the Emulsion, at a Temperature Below 40° C.

[0099] Tocopherol Acetate between 0.1 and 1%, Pyridoxine between 0.01 and 0.05%, Vitamin A Palmitate between 0.01 and 1%, d-Panthenol between 0.1 and 1%, Citric Acid between 0.1 and 0.5%, Zinc Gluconate between 0.1 and 1%, Trisodium Citrate between 1 and 2.5%, Water 5%.

Example 8: Protective Mist for Skin Exposed to Blue Light from Electronic Devices

[0100] Water qsp. 100%, Kaempferitrin 2%, Preservatives 0.6%, Glycerin 7%, Hexylene Glycol 3.0%, Carbomer 0.5%, Tetra sodium EDTA 0.2%, Sodium Hyaluronate 0.1%, NaOH 0.5%, Preservative+fragrance qs.

Example 9: Comparison of the Effects of Astaxanthin and Myricitrin on the Release of IL31 by Primary Keratinocytes Exposed to the Combined Effects of Blue Light and of benzo[a]pyrene (BaP)

[0101] Astaxanthin (dihydroxy-3,3′ dioxo-4,4′ (3-carotene) is a carotenoid synthesized in particular by microalgae. This molecule absorbs wavelengths comprised between 400 nm and 500 nm.

[0102] Benzo[a]pyrene (CAS 50-32-8) is one of the most toxic polycyclic aromatic hydrocarbons (PAHs).

[0103] The cell treatments were carried out as follows. [0104] a) Cell type: primary keratinocytes from human epidermis (39-year-old donor, ref: KER110, batch KER110049, Biopredic) [0105] b) Culture conditions: 96-well plate, 37° C., 5% CO.sub.2, seeding 20,000 cells/cm.sup.2 [0106] c) Culture medium: KSFM (Gibco ref: 10144892)+1% penicillin/streptomycin [0107] d) Treatment with the compounds: performed 24 hours after seeding, [0108] e) Incubation time with the compounds: 24 hours of contact [0109] f) Concentrations of the compounds tested: myricitrin and astaxanthin at 20 μM and 50 μM [0110] g) Stress condition: performed following the treatment with the compounds; exposure to irradiation (blue light; 40 J/cm.sup.2; 450 nm); followed by 24 hours of treatment with 20 μM of benzo[a]pyrene (CRM40071, Sigma-Aldrich) in culture medium.

[0111] The analyses were performed according to the following procedure.

[0112] Control batch: no treatment other than the renewal of the culture medium

[0113] Samples: [0114] Culture media [0115] Cell pellets

[0116] Endpoints/readout: [0117] IL-31 assay, using the culture media (Human IL-31 ELISA MAX Deluxe; Biolegends) [0118] Assay of total proteins, using the cell pellets (Bradford Method)

[0119] Results: IL31 Assay

[0120] In FIG. 4, the results are expressed as IL-31 concentration normalized by the amount of total cell proteins.

[0121] In FIG. 5, a release inhibition value (%) was calculated for the experimental groups, using the following formula:

% IL31 Release Inhibition: % Batch X=(IL31 Stress−IL31 Batch X)/(IL31 Stress−IL31 Control)×100

[0122] As a reference, the control batch was examined at maximum efficiency (100%) and the stress group (Blue light+BaP) at minimum efficiency (at 0%):