Cosmetic Composition

Abstract

A cosmetic composition is provided that includes a carrier and a patchouli stem extract, particularly an extract from spent patchouli stem, which provides antioxidant, anti-elastase, anti-hyaluronidase, and anti-tyrosinase activity.

Description

[Technical field]

[0001] The present invention relates to a cosmetic composition comprising a patchouli stem extract. [Background technology]

[0002] Patchouli (Pogostemon cablin) is a species of plant in the Lamiaceae family, commonly referred to as the "mint" or "laminaceous" family. The plant grows as a bushy herb with erect stems reaching a height of about 75 cm and producing small pale pink-white flowers. It is native to tropical regions of Asia.

[0003] Patchouli's heavy, strong scent has been used in perfumery for centuries, and more recently in incense, insect repellents, and alternative medicine. Pogostemon cablin, among other members of the Pogostemon genus, is commonly cultivated for its essential oil, known as patchouli oil. Patchouli grows well in warm to tropical climates. The seed-producing flowers are extremely fragrant and bloom in late fall. The tiny seeds may be harvested for planting, but they are quite delicate and easily crushed. Cuttings from the mother plant can also be rooted in water to produce further plants.

[0004] Extraction of patchouli's essential oil is usually accomplished by steam distillation of the dried leaves and stems, which requires rupturing its cell walls by steam burning, light fermentation, or drying. The leaves are harvested several times a year and, when dry, may be exported for distillation.

[0005] The main chemical component of patchouli oil is patchoulol, a sesquiterpene alcohol. Other important components include germacrene-B, and norpatcholenol. Overall, about 140 chemicals have been identified in patchouli, including terpenoids, phytosterols, flavonoids, organic acids, lignins, alkaloids, glycosides, alcohols, and aldehydes (Swamy et al. in Molecules 2015, 20, 8521-8547). The volatiles consist mainly of patchouli alcohol, α-patcholene, β-patcholene, α-bulnesene, seichelen, norpatcholenol, pogostone, eugenol, and pogostol. The non-volatile substances include acacetin, apigenin, licochalcone A, ombuin, rhamnetin, retsucin, acteoside, agastacoside, isocrenatoside, tilianin, rubusoside, tri- and sesquiterpenes.

[0006] Patchouli is widely used in fine fragrances and flavorings and is an important ingredient in East Asian incense. Patchouli leaves have been used to make an herbal tea. In some cultures, patchouli leaves are eaten as a vegetable or used as a seasoning. Patchouli has also been described as an insect repellent against Formosan termites. Summary of the Invention

[0007] The objective of the present invention is to add value to patchouli stems which have been used to produce patchouli oil. This is accomplished by the cosmetic compositions and methods of the present invention. In a first aspect, the present invention provides a cosmetic composition comprising a carrier and at least one active cosmetic ingredient, the first active cosmetic ingredient comprising patchouli stem extract.

[0008] In contrast to essential oils, which are obtained by hydrodistillation or steam distillation, patchouli stem extract is obtained by extracting patchouli stems with a liquid solvent. The patchouli stem extract of the present invention is completely natural and odorless. Surprisingly, it has been found that the patchouli stem extract of the present invention has excellent skin care and anti-aging properties. In particular, it has been found to exhibit antioxidant activity, anti-elastase activity, anti-hyaluronidase activity, and anti-tyrosinase activity. Thus, in a particular embodiment, the cosmetic composition of the present invention is a skin care composition, in particular an anti-aging composition. To this end, the carrier must be a dermatologically acceptable carrier.

[0009] A particularly suitable extract is an aqueous extract of patchouli stem.Accordingly, in a preferred embodiment of the present invention, the first active cosmetic ingredient comprises an aqueous extract of patchouli stem. Another particularly suitable extract is an alcohol extract of patchouli stem.Accordingly, in a preferred embodiment of the present invention, the first active cosmetic ingredient comprises an alcohol extract of patchouli stem.Ethanol is a particularly preferred solvent. In a particularly preferred embodiment, Patchouli stems - and in particular exhausted Patchouli stems - are extracted with an alcohol-water mixture. Ethanol-water mixtures are especially preferred. The aqueous extract may be obtained by extracting the stems of patchouli with pure water. Optionally, the water may contain additives, such as those to adjust the pH.

[0010] Surprisingly, it has been found that even an extract of spent patchouli stem provides an active cosmetic ingredient having the above-mentioned beneficial effects.Accordingly, in a specific embodiment of the present invention, the first active cosmetic ingredient comprises an extract of spent patchouli stem.

[0011] Throughout this application, the term "spent patchouli stem" refers to patchouli stems that have previously been processed by steam distillation, and thus refers to the residue of the roots after the essential oil has been removed. In a specific embodiment, the present invention relates to an aqueous and / or alcoholic extract of spent patchouli stems, which allows adding value to a "waste" product.

[0012] In a further aspect, the present invention relates to a method for preparing an active cosmetic ingredient, said method comprising extracting patchouli stems. Extracting means that patchouli stems are treated with a solvent or a mixture of solvents. The solvent or solvents may also contain additives. For example, patchouli stems may be subjected to aqueous extraction, acidic extraction, enzymatic extraction, ultrasonically assisted aqueous extraction, pressurized aqueous extraction or ethanol extraction.

[0013] Preferably, the patchouli stems are reduced to smaller pieces prior to extraction, in particular by cutting and / or grinding. The patchouli stems may be washed prior to extraction. Washing the stems has been found to reduce the color of the extract, especially in water. Preferably, the patchouli stems are extracted with water, ethanol or a mixture thereof. Extraction with 70% ethanol in water, optionally containing one or more additives, has been found to give the best results. Suitable additives include, but are not limited to, acids, bases, buffers, salts and / or co-solvents. In particular, the pH of the extraction solvent may be adjusted by the addition of an acid (e.g., H2SO4 or citric acid) or a base (e.g., NaOH).

[0014] The extraction may be carried out at room temperature or at an elevated temperature, for example at a temperature of about 40°C, about 60°C or about 80°C. The patchouli stem extract may be purified, for example, by filtration (eg, on a KDS15 filter), activated charcoal treatment, and / or sterile filtration. The patchouli stem extract may also be concentrated. To improve the solubility of the extract during concentration, a solvent, such as 1,3-propanediol, can be added to the extract before concentration.

[0015] In a particular embodiment, the method of the present invention comprises the steps of: (i) providing spent patchouli stems; and (ii) extracting the spent patchouli stems. Thus, in conclusion, the compositions of the present invention provide stimulation of sebum production, stimulation of antioxidant properties, and potent inhibition of wound healing via regulation of keratinocyte migration. The compositions of the invention may therefore be used in skin care, scalp care and / or body care compositions, in particular in serums for dry skin, anti-ageing serums, anti-ageing night and day creams, dry anti-dandruff products, dry scalp lotions or body care lotions. In a further aspect, the present invention relates to the use of an extract from the stem of Patchouli in skin care, which makes it possible to take advantage of the positive effects of the extract of Patchouli stem described above.

[0016] In a specific embodiment, an aqueous extract and / or an extract from spent patchouli stems is used. Patchouli stem extract is particularly advantageously used in anti-aging products, anti-dandruff products or for dry skin and / or scalp. In a further aspect, the present invention relates to a method for stimulating sebum production, stimulating antioxidant properties and / or inhibiting wound healing by applying an inventive cosmetic composition to human skin.

[0017] The present invention is further illustrated by the following non-limiting examples. Example 1: Preparation of Patchouli Stem Extract Dried steam distilled patchouli stems were crushed to obtain a powder. 25 g of the powder was extracted in 500 g of ethanol (70% in water) at room temperature with stirring for 30 minutes. The extract (approximately 420 g) was filtered through an Eaton KDS15 filter (123 cm 2 The filtrate was concentrated 5.2 times by evaporation under vacuum to give about 80 g of concentrated product. One batch of concentrated product (Batch A) was used as is. This unbleached product was black in color. The second batch of concentrated product (Batch B) was filtered through an activated carbon filter (24 cm 2 This bleaching process effectively reduced the color. It reduced the dry matter by almost a factor of two. The two batches were finally filtered using Eaton S60 filters (24 cm 2 ) and filtered through a sterile PES filtration unit (VWR), and the final product was stored without preservatives.

[0018] The characteristics of the two batches are shown in the table below: [Table 1]

[0019] For comparison of different solvents, extractions were also performed with water, 100% ethanol, 70% ethanol and 50% ethanol. Colour (Gardner) and dry matter content were measured in the raw extracts. The results are shown below. [Table 2] Essentially no differences in colour or even dry matter content were observed (except for 100% ethanol), although the yields of dry matter content were generally quite low.

[0020] Example 2: Analysis of patchouli stem extract Dried steam-distilled patchouli stems were ground to obtain a powder. 75 g of the powder was extracted in 550 g of ethanol (70% in water) at 20°C for 60 min with stirring. The extract was filtered through a 0.7 μm cellulose filter, concentrated five times to remove the ethanol, and finally filtered through a 0.35 μm cellulose filter. The extract was then freeze-dried.

[0021] Fractionation of crude extracts by centrifugal partition chromatography (CPC) The crude extract (1.078 g) was dissolved in 30 ml of a biphasic solvent system consisting of methyl tert-butyl ether (MTBE), acetonitrile and water in a ratio of 3:3:4 (v / v). Centrifugal partition chromatography (CPC) was performed using a FCPE300® apparatus (Rousselet Robatel Kromaton) with a 303 ml column, a column rotation speed of 1200 rpm and a flow rate of 20 ml / min.

[0022] Isocratic elution of the mobile phase (lower phase of the two-phase solvent system) was performed for 75 min in ascending mode (initial flow rate ramp from 0 to 20 ml / min for 5 min). The column was finally pushed out by switching the mode selection valve for 20 min. The CPC chromatogram was monitored at 220 nm. Fractions of 20 ml were collected throughout the experiment and combined according to the thin-layer chromatography (TLC) profile. TLC was performed on precoated silica gel 60 F254 Merck plates with the mobile solvent system EtOAc / toluene / formic acid / acetic acid (70 / 30 / 11 / 11; v / v) and visualized under UV light at 254 nm and 360 nm, and revealed by spraying the dried plates with 50% H2SO4 and vanillin followed by heating. As a result, 16 subfractions were collected.

[0023] NMR analysis and identification of major metabolites An aliquot of each fraction from F1 to F16 (up to approximately 20 mg) was dissolved in 700 μl of DMSO-d6 and cryo-detected at 298 K on a Bruker Avance AVIII-600 spectrometer equipped with a TXI cryoprobe (Karlsruhe, Germany). 13 The spectra were manually phased, baseline corrected, and calibrated to the central resonance of DMSO-d6 (δ 39.80 ppm) using TOPSPIN 3.2 software (Bruker). 13All absolute intensities of the C NMR signals were automatically collected and binned across the spectra of the fraction series using a locally developed computer script. The resulting table was imported into PermutMatrix version 1.9.3 software (LIRMM, Montpellier, France) for hierarchical clustering analysis (HCA). 13 The C chemical shift clusters were visualized as a dendrogram on a two-dimensional map. For the identification of metabolites, 13 The C chemical shift clusters were manually submitted to the structural search engine of the database management software ACD / NMR Workbook Suite 2012 software (ACD / Labs, Ontario, Canada), which contains structures and predicted chemical shifts of low molecular weight natural products (n ≈2950 in March 2018). In parallel, a literature search was performed to obtain the names and chemical structures of the maximum number of metabolites already reported in the species Pogostemon cablin (n ≈70). Additional 2D NMR experiments (HSQC, HMBC, and COSY) were performed on fractions containing putatively identified compounds at the end of the dereplication process to confirm the molecular structures proposed by the database.

[0024] The following major metabolites were identified: - Verbascoside - Apigenin-7-O-glucuronide - Succinic acid - Glucosyl-cytosporone V (2 isomers) - Lactic acid - Colin - α-D-fructose - β-D-fructose - α-D-glucose - β-D-glucose - β-D-fructopyranose - 7,3'-Dimethyleriodictyol - Pachypodium - Cytosporon V - p-Hydroxybenzoic acid - Protocatechuic acid - Syringaresinol - Vanillin - Rhamnocitrine - Luteolin-7-O-glucuronide - Hydroxymethylglutaric acid - 12-Hydroxyjasmonic acid - Caffeic acid derivatives - Apigenin-7-saccharide

[0025] The composition of the CPC fractions was as follows (Maj=major; Med=medium; Min=minor): [Table 3]

[0026] Example 3: Biological activity screening The two batches A and B from Example 1 were tested for antioxidant activity (DPPH), anti-elastase activity, anti-hyaluronidase activity, and anti-tyrosinase activity. The results are shown in the following table: [Table 4] The undecolorized batch A clearly showed higher biological activity. The decolorized batch B displayed only slight antioxidant activity but showed similar anti-tyrosinase activity as batch A.

[0027] Example 4: Wound healing Normal human epidermal keratinocytes (NHEK) were seeded at 200,000 cells per well precoated with type I collagen in 12-well culture plates in the presence of Keratinocyte Growth Medium (KGM, Lonza) supplemented with growth factors such as hydrocortisone, transferrin, epinephrine, bovine pituitary extract (BPE), recombinant human epidermal growth factor (rhEGF) and insulin. At confluence, cells were preincubated overnight with 0.5% (v / v) concentration of patchouli stem extract from Example 1 or 1 ng / ml heparin-binding epidermal growth factor (HB-EGF) (positive control) in Keratinocyte Basal Medium without supplements. After this preconditioning step, the cell monolayer attached to the bottom of the well was scratched with a P200 sterile cone and subsequently washed twice with phosphate-buffered saline (PBS). NHEKs were then restimulated with 0.5% patchouli extract or 1 ng / ml HB-EGF in basal medium for 8 hours.

[0028] The wound healing process was monitored at T0 and T2 using an inverted optical microscope (Zeiss). 8h After image analysis, the percentage of scratch closure was determined in comparison to the untreated condition. It was found that patchouli stem extract significantly inhibited wound healing in the NHEK scratch assay by about -50%, while HB-EGF produced an enhancement of about +60%. Inhibition of wound healing is particularly important in conditions involving hyperproliferation of keratinocytes, such as hyperkeratosis or dry dandruff.

[0029] Example 5: Antioxidant effect NHEKs were seeded in black glass-bottom plates at 20,000 cells per well precoated with collagen type I. Cells were incubated for 24 h at 37 °C and 5% CO2. The next day, cells were incubated for 24 h with patchouli stem extract from Example 1 at a concentration of 0.5% (v / v) or 200 μM resveratrol (positive control). After this preincubation, cells were incubated with 50 μM dichloro-dihydro-fluorescein (DCFH) probe for 30–45 min. Cells were then rinsed twice with PBS and incubated in PBS alone or with PBS containing 5 mM tert-butyl peroxide (TBP) to induce oxidative stress. Fluorescence readings were taken every 10 min for 1 h, with excitation at 488 nm and emission at 525 nm. Patchouli stem extract was found to reduce the production of reactive oxygen species (ROS) by -28%, proving its antioxidant properties.

[0030] Example 6: Regulation of sebum production Human sebocytes were seeded in 96-well plates (50,000 cells / well) and cultured for 24 hours in keratinocyte serum-free medium (SFM) supplemented with 25 μg / ml gentamicin. The medium was then removed and replaced with the patchouli stem extract of Example 1 at a concentration of 1% (v / v) or with 1 μM of the reference ormacostat grasaretil. The cells were pre-incubated for 4 hours. Then, an adipogenic mixture containing vitamin C, vitamin D3, insulin and calcium (without androgens) was added and the cells were incubated for 7 days. After 3 days of incubation, half of the medium was removed and the treatment was renewed (adipogenic mixture stimulation included). An unstimulated control condition was performed in parallel. At the end of the incubation, the cells were rinsed, fixed and permeabilized. The lipid droplets contained in the cells were then labeled using a specific Bodipy® fluorescent lipid probe that mainly labels neutral lipids. In parallel, the cell nuclei were stained using a Hoechst33258 solution. Image acquisition was performed using an IN Cell analyzer™ 2200. Five pictures were taken per well (×20 objective). Labeling was quantified by fluorescence intensity measurements normalized to the total number of cells. Patchouli stem extract was found to significantly induce sebum production from sebocytes by +39% under lipogenic stimulation. Stimulation of sebum production is particularly useful for treating dry skin and restoring the skin barrier and skin permeability.

Claims

1. A cosmetic composition comprising a carrier and at least one active cosmetic ingredient, the active cosmetic ingredient comprising a depleted patchouli stem extract; Spent patchouli stems are patchouli stems that have previously been processed by steam distillation. The spent patchouli stem extract is extracted from spent patchouli stem with water or a mixture of alcohol and water at a temperature between room temperature and 80°C. The cosmetic composition.

2. The cosmetic composition according to claim 1 , wherein the cosmetic composition is a skin care, scalp care or body care composition.

3. 3. The cosmetic composition according to claim 2, wherein the cosmetic composition is a serum for dry skin, an anti-aging serum, an anti-aging night or day cream, a dry anti-dandruff product, a dry scalp lotion or a body care lotion.

4. A cosmetic composition according to any one of claims 1 to 3, wherein the active cosmetic ingredient comprises an aqueous and / or alcoholic extract of exhausted patchouli stems.

5. A cosmetic composition according to any one of claims 1 to 4 for stimulating sebum production, for suppressing the generation of reactive oxygen species and / or for inhibiting wound healing.

6. A cosmetic composition according to any one of claims 1 to 4 for stimulating sebum production.

7. A cosmetic composition according to any one of claims 1 to 4 for inhibiting wound healing.

8. 1. A method for preparing an active cosmetic ingredient, comprising: (i) providing spent patchouli stems; and (ii) extracting the spent patchouli stems with water or a mixture of alcohol and water at a temperature between room temperature and 80° C.; wherein the spent patchouli stems are patchouli stems that have previously been treated by steam distillation.

9. 9. The method according to claim 8, wherein the extraction is carried out using water and / or ethanol.

10. 1. Use of an extract from spent patchouli stems in skin care, wherein the spent patchouli stems are patchouli stems that have been previously treated by steam distillation, and the extract from spent patchouli stems is extracted from spent patchouli stems with water or a mixture of alcohol and water at a temperature between room temperature and 80°C.

11. A cosmetic method for stimulating sebum production and / or suppressing the production of reactive oxygen species by applying a cosmetic composition according to any one of claims 1 to 4.

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