KR20200006515A - Anti-inflammatory, anti-wrinkle, whitening cosmetic composition comprising the extract or fraction of Zanthoxylum piperitum DC. as an active ingredient and preparation method of the same - Google Patents

Abstract

The present application relates to a Zanthoxylum piperitum extract or a fraction thereof, a method for preparing the same, and an anti-inflammatory, anti-wrinkle and whitening functional cosmetic composition containing the same.

Description

Anti-inflammatory, anti-wrinkle, whitening cosmetic composition comprising the extract or fraction of Zanthoxylum piperitum DC. as an active ingredient and preparation method of the same}

The present application relates to a functional cosmetic composition and a method for producing the same containing the extract of the bark or its fraction as an active ingredient.

Inflammatory reactions are mechanisms for repairing and repairing damaged areas when invasion results in any organic changes, such as physical actions, chemicals, or bacterial infections, but continuous inflammatory reactions promote mucosal damage. Some lead to diseases such as cancer. Macrophage cells are known to be involved in homeostasis, which is involved in various host reactions such as acquired immunity as well as acquired immunity. In addition, macrophages produce nitric oxide (NO) and cytokine during inflammatory reactions, and play an important role in the early defense of infection. Do it. Among them, NO has various physiological functions such as body defense function, signaling function, neurotoxicity, and vasodilation. In general, NO formation plays an important role in killing bacteria or removing tumors, but it is excessively produced by iNOS in inflammatory state. NO has been known to promote inflammatory reactions such as vascular permeability and edema, as well as to promote inflammation and to intensify inflammation (Yoon et al., Korean J. Food Sci. Technol. Vol 39, No. 4, 464-469, 2007) NO production inhibition is used as an indicator of inflammation inhibition.

Melanin (Melanin) is a pigment that determines the eyes, hair, or skin color of the body and is synthesized in the process of protecting cells from external stimuli such as ultraviolet rays. When the skin is irritated by UV light, cyclic adenosine monophosphate (cAMP) / protein kinase A (PKA) binds to the melanocortin 1 receptor (MC1R), which is present in the melanocytes of the epidermis, and the α-melanocyte stimulating hormone (α-MSH) secreted by the pituitary gland. The expression of microphthalmia-associated transcription factor (MITF) is increased by the pathway. This increases the activity of tyrosinase, tyrosinase related protein-1, -2 (TRP-1, -2), which plays an important role in melanin synthesis, and induces melanin synthesis. This action protects our skin from various skin diseases such as skin cancer that can occur from ultraviolet rays. However, if this action is excessive, it may cause side effects such as blemishes and freckles due to pigmentation (Kwon et al., Journal of Life Science, Vol 24, No 9, 946-951, 2014).

As human skin ages, endogenous aging occurs because the secretion of various hormones that regulate metabolism decreases, and the function and activity of immune cells and skin cells decreases, reducing the biosynthesis of immune proteins and bioconstituent proteins necessary for the living body. And external aging is known that wrinkles caused by photoaging by various pollutants and ultraviolet rays, decreased elasticity, blemishes due to increased skin dryness, freckles and black mushrooms are produced. Collagen, a major component of skin cells, accounts for about 70-80% of the dry weight of the dermis, of which type I collagen accounts for 80% of total collagen and type III collagen 15%. In general, the synthesis of type I collagen and the activity of MMP-1, its degrading enzymes, in the skin maintain the proper balance and maintain the elasticity of the skin. If this balance is broken by endogenous aging and exogenous aging factors, skin wrinkles are produced. (Youn et al., Korean J. Food Preserv 19 (3), 393-399, 2012).

Zanthoxylum piperitum DC. Is a deciduous leaf family belonging to the Zanthoxylum of Rutaceae, which is native to Korea, China, Japan, and Northeast Asia. These species contain various spicy, essential oils and fats and oils. It has been widely used as the oldest traditional spice, medicinal and oil-making in Northeast Asia since ancient times (Kim et al., 2000). Chopi is an essential oil ingredient that has a sour taste and a characteristic citrus-like scent. Since ancient times, Chinese pepper has been used as the best spice among five incense (chochu, cinnamon, fennel, dermis, cloves, etc.). In Japan, it is called Japanese pepper and has been widely used as a spice or medicinal ingredient that gives a distinctive flavor to Japanese cuisine. In Korea, it is used as a spice that grinds fruit and puts it in Chueotang, Maengtang, Bosintang, etc., and the old literature used Chopi as a spice of kimchi before red pepper was used in Kimchi (Na et al., Changwon National University). , 2004). The main components of the bark tree are known as quercetin, afzelin, quercitrin, hyperoside, hesperidin, etc. (Hur et al., Natural Product Sciences, 7 (1), 23-26, 2001). Hatano et al., Phytochemistry, 65 (18), 2599-2604, 2005), hydroxy-sanshool, bugeanol, and the like (Matthias et al., J. Agic. Food. Chem, 62, 2479-2488, 2014) .

Republic of Korea Patent No. 10-1303950 discloses a composition for the prevention and treatment of bone diseases containing the extract of the bark wood as an active ingredient, Republic of Korea Patent No. 10-0480972 is a dioxin-like similar to the extract of the bark wood as an active ingredient An antagonistic composition for a substance is disclosed. On the other hand, the Republic of Korea Patent Publication No. 10-20160036198 discloses a sunscreen cosmetic composition containing the extract as an active ingredient, but the anti-inflammatory, whitening, wrinkle improvement function containing the extract or fractions thereof as an active ingredient There is no research on the functional cosmetic composition which has at the same time. Under this background, the present inventors have completed the present application by earnestly researching efforts to develop anti-inflammatory, anti-wrinkle or whitening functional cosmetics containing the extract of the bark or its fraction as an active ingredient.

It is an object of the present application to provide a method for preparing the bark tree extract.

Another object of the present application is to provide a method for producing the bark fraction.

Another object of the present application is to provide a bark tree extract and a bark tree fraction.

Another object of the present application is to provide an anti-inflammatory, anti-wrinkle or whitening functional cosmetic composition containing any one or more substances selected from the group consisting of bark tree extract and bark tree fraction as an active ingredient.

Other objects and advantages of the present application will become apparent from the following detailed description taken in conjunction with the appended claims and drawings. Content not described herein is omitted because it can be sufficiently recognized and inferred by those skilled in the art or similar fields of the present application.

Hereinafter, the content of the present application will be described in detail. In addition, description and embodiment of one embodiment disclosed by this application can be applied also to description and embodiment of another aspect about common matter. In addition, all combinations of the various elements disclosed in the present application are within the scope of the present application. In addition, the scope of the present application is not to be limited by the specific description described below.

In order to achieve the object of the present application, the present application provides a method of preparing the bark extract as an aspect. The bark tree may be used in a dried form, and extracted with an organic solvent including ethanol, benzene, and acetone to obtain an extract in a concentrated manner using a device such as a vacuum suction filter. In the present invention, "extract" means an extract obtained by pulverizing the bark tree, hot water extraction method, ethanol extraction method and the like. The bark tree part may include various parts of the plant, such as leaves, branches, and fruits, but is not limited thereto. The bark leaves may use a weight of 100 to 200 g, preferably 130 to 180 g, most preferably 150 g is used. The bark branches and fruits may use a weight of 200 to 700g, preferably 300 to 600g, more preferably 400g. The concentration of ethanol that can be used to obtain the extract of the bark tree such as leaves, branches, fruits of the bark tree may be 30 to 70% (V / V), more preferably 40 to 60% (V / V). Most preferably 50% (V / V) is used.

In another aspect, the present application provides a method of preparing the bark fraction by further using a fractionation solvent in addition to the bark extract. The fractionated solvent may use hexane, chloroform, ethyl acetate, butanol, distilled water, but is not limited thereto. The fractionation layer can be obtained by using an experimental technique for separating a mixture such as chromatography. Chromatography is normal silica gel Column chromatography packed with, sephadex LH-20 packed column chromatography can be used selectively or sequentially depending on the desired fraction.

According to one aspect of the present application, the present application provides an anti-inflammatory, anti-wrinkle and whitening functional cosmetic composition containing the non-cytotoxic extract of the bark tree, fractions and the active ingredient prepared by the above method. Cytotoxicity can be performed by adding a reagent such as Thiozolyl Blue Tetrazolium Bromide (MTT) and measuring it using a microplate counter, leakage assay, or Neutral red assay.

In the present application, the content of the bark extract or fractions thereof contained in the cosmetic composition may be used in an amount of 0.01 to 30.0 wt% based on the total weight of the cosmetic composition. Preferably 5 to 20% by weight can be used and most preferably 10% by weight is used. If it contains a ratio less than the above weight% range, it is difficult to substantially exert anti-inflammatory, anti-wrinkle, whitening effect, and if it contains more ratio than the above, it is not economical.

In the present application, the functional cosmetic composition containing the extract of the vinegar or fractions thereof as an active ingredient is a lotion, skin lotion, skin softener, skin toner, astringent, lotion, milk lotion, moisturizing lotion, nutrition lotion, massage cream, nutrition cream , Moisturizing cream, hand cream, essence, nutrition essence, pack, soap, shampoo, cleansing foam, cleansing lotion, cleansing cream, body lotion, body cleanser, emulsion, characterized in that any one selected from the group consisting of.

As used herein, the term "anti-inflammatory" is a generic term for a disease whose main lesion is inflammation, but is not limited thereto, and preferably includes atopic dermatitis, psoriasis, allergic dermatitis, contact dermatitis and eczema. do.

The present application has an excellent effect of providing an extract or fractions thereof and an anti-inflammatory, anti-wrinkle and whitening functional cosmetic composition containing them.

1 is a schematic diagram showing the manufacturing process of the present invention bark leaf extract and organic compounds.
Figure 2 is a schematic diagram showing the manufacturing process of the present invention bark tree branch extract and organic compounds.
Figure 3 is a schematic diagram showing the manufacturing process of the present invention bark fruit extract and organic compounds.
Figure 4 is a graph showing the results of the anti-inflammatory activity according to the concentration of the extract of the bark of the present invention.
Figure 5 is a graph showing the results of the anti-inflammatory activity according to the concentration of the extract of the bark of the present invention.
Figure 6 is a graph showing the results of the anti-inflammatory activity according to the concentration of the extract of the bark fruit of the present invention.
7 is a graph showing the results of the anti-wrinkle activity of the extract of the bark of the present invention.
8 is a graph showing the results of the anti-wrinkle activity of the extract of the fruit bark of the present invention.
Figure 9 is a graph showing the results of the whitening activity of the extract of the bark of the present invention.
10 is a graph showing the results of the whitening activity of the extract of the bark of the present invention.
Figure 11 is a graph showing the results of the whitening activity of the present invention bark fruit extract.
12 is a graph showing the results of the anti-inflammatory activity of the organic compound 3 isolated from the leaves and berries of the present invention bark.
13 is a graph showing the results of the whitening activity of the organic compound 3 isolated from the leaves and berries of the present invention bark.
14 is a graph showing the results of the anti-inflammatory activity of the organic compound 4 isolated from the bark of the present invention.

Hereinafter, the present invention will be described in more detail through examples according to the present application. However, the following examples of the present application are merely examples of the present application. These examples are intended to illustrate the present invention in more detail, and it is obvious to those skilled in the art that the scope of the present invention as set forth in the appended claims is not limited by these examples. something to do.

Example 1 Obtaining Cedar Bark Leaf Extract of the Present Invention and Organic Compounds 1,2,3

150g of dried vine leaves were extracted using 50% ethanol (V / V). The extract was concentrated only by the filtrate using a vacuum suction filter to obtain a vine leaf extract.

The bark leaf extract (21 g) was suspended in 100% distilled water and fractions were obtained in the following order: hexane, chloroform (190 mg), ethyl acetate (1.5 g), butanol, distilled water. Among them, ethyl acetate fractionation layer was subjected to column chromatography packed with normal silica gel to obtain organic compound 1 (358 mg, Chemical Formula 1). The results of 1H & 13C-NMR data of organic compound 1 are shown in Table 1. Next, sephadex LH-20-filled column chromatography was performed to obtain organic compound 2 (16 mg, Chemical Formula 2). The results of 1H & 13C-NMR data of organic compound 2 are shown in Table 2. Chloroform fractional layer was subjected to column chromatography packed with normal silica gel to obtain organic compound 3 (9.7 mg, Chemical Formula 3), and the results of 1H 13C-NMR data of organic compound 3 are shown in Table 3 (FIG. 1). .

Example 2 Preparation of Extract of Organic Bark Tree Branch and Organic Compound 4 of the Present Invention

400 g of dried bark branches were extracted using 50% ethanol (V / V). The extract was concentrated only by the filtrate using a vacuum suction filter to obtain a bark branch extract.

Succulent branch extract (12 g) was suspended in 100% distilled water and fractions in the order of hexane, chloroform, ethyl acetate, butanol in order to obtain a fraction. Among them, the hexane fractionation layer was subjected to column chromatography filled with normal silica gel and sephadex LH-20 to obtain organic compound 4 (1.0 mg, Chemical Formula 4), and 1H & 13C-NMR data of organic compound 4 were obtained. The results are shown in Table 4 (FIG. 2).

[Example 3] Preparation of the present invention bark fruit extract and organic compounds 1,3,5

400g of dried fruit trees were extracted using 50% ethanol (V / V). The extract was concentrated only by the filtrate using a vacuum suction filter to obtain an extract of the bark fruit.

Succulent fruit extract (28g) was suspended in 100% distilled water and fractions in the order of hexane, chloroform, ethyl acetate, butanol, distilled water to obtain a fraction. The hexane fractionation layer was subjected to column chromatography packed with normal silica gel to obtain organic compound 3 (158.8 mg, formula 3). Ethyl acetate fractionation layer obtained organic compound 5 (85.1mg, Formula 5) by the precipitation experiment, the results of 1H & 13C-NMR data of organic compound 5 are shown in Table 5. In addition, Ethyl acetate fractionation layer was subjected to column chromatography packed with normal silica gel and sephadex LH-20 in order to obtain an organic compound 1 (49.5mg, Formula 1) (Fig. 3).

Experimental Example 1 Anti-inflammatory Activities of Extracts from Bark Trees and Organic Compounds 3 and 4

In order to measure the anti-inflammatory activity of the leaves, branches and fruit extracts of the bark tree tree obtained from Examples 1, 2, and 3, 4, the extracts were artificially treated with LPS, an inflammation-inducing substance, in RAW 264.7 cells. And organic compounds were treated to evaluate whether they have an anti-inflammatory effect. That is, 2 x 10 ⁵ cells were dispensed per well in a 96-well plate, and the cells were cultured in 37 ° C., 5% CO ₂ incubator for 24 hours. The cells were incubated overnight and then exchanged with fresh medium. LPS (1㎍ / mL) and each bark extract were at concentrations of 0.5, 1,2% (W / V), and organic compound 3 was 10, 50, 100㎍. At a concentration of / mL, the organic compound 4 was incubated for 24 hours by treating at a concentration of 5, 10, 50㎛ / mL. After incubation, the culture supernatant was collected and centrifuged at 13,000 rpm for 3 minutes to collect only the supernatant, and then reacted 1: 1 with Griess reagent and the absorbance was measured at 570 nm.

As a result of measuring the NO production inhibitory effect after extracting 40% butylene glycol (BG), at the 2% concentration (W / V) of each extract, the bark leaves, eggplants, fruit extracts were about 43% (Fig. 4), 50% (FIG. 5) 66% (FIG. 6), organic compound 3 (FIG. 12) at about 92% at 100 μg / mL, and organic compound 4 at about 96% at 50 μg / mL (FIG. 14). Showed high NO production inhibitory efficacy.

Experimental Example 2 Anti-Wrinkle Activity of Extracts from Bark Trees

Type I procollagen biosynthesis in fibroblasts was measured in order to measure the anti-wrinkle activity of the leaves and fruit extracts of the bark tree obtained in Examples 1 and 3. To this end, fibroblasts were inoculated in 48 well plates at a concentration of 5x10 ⁴ cells / mL and the cells were grown. DMEM medium containing 10% serum was used, and 24 hours after inoculation, test samples were added to the medium at 10, 50, and 100 µg / mL and incubated in a 37 ° C., 5% CO ₂ incubator for 1 day. Procollagen type I C-peptide was measured using the supernatant obtained by transferring the medium into the tube and centrifuging at 15,000 rpm for 30 seconds.

As a result, the extract of the bark tree branch extract of about 280% (Fig. 7), the bark fruit extract of about 82% (Fig. 8) at the concentration of 100 ㎍ / mL after 50% ethanol (EtOH) extraction showed an increased collagen synthesis efficacy Confirmed.

Experimental Example 3 Whitening Activity of Bark Extract and Organic Compound 3

Inhibition of melanin formation was carried out to measure the whitening activity of the leaves, branches and fruit extracts and the organic compounds 3 obtained in Examples 1, 2 and 3 above. To this end, murine melanoma (B-16 F1) cells were inoculated with 1 × 10 ⁵ per well in a 6-well plate with DMEM medium containing an appropriate amount of fetal bovine serum (FBS), followed by 5% CO ₂ at 37 ° C. Were cultured until at least about 80% was attached to the bottom of the well. After incubation, the medium was removed, and the sample was replaced with a medium diluted to an appropriate concentration, and then incubated for 5 hours under 5% CO ₂ , 37 ° C. The cells from which the medium was removed were washed with PBS (phosphated buffer saline) and treated with trypsin to recover the cells. The collected cells were counted using a hematocytometer, centrifuged at 5,000 to 10,000 rpm for 10 minutes, and the supernatant was removed to obtain pellets. The cell pellet was dried at 60 ° C., and then added 100 μl of 1M sodium hydroxide solution containing 10% DMSO or an appropriate amount of cell lysis buffer to obtain intracellular melanin in a 60 ° C. thermostat. Using the solution, the absorbance was measured at 405 nm using a microplate meter to determine the amount of melanin per cell constant or the amount of melanin per protein.

As a result, after extracting with 50% ethanol (EtOH), at the concentration of 100㎍ / mL, the bark leaf extract is about 27% (Fig. 9), the bark tree extract is about 18% (Fig. 10), the bark fruit extract It showed an effect of inhibiting melanin production of about 25% (FIG. 11). In addition, organic compound 3 confirmed the effect of inhibiting melanin production of about 20% (FIG. 13) at a concentration of 100 µg / mL.

Experimental Example 4 Investigation of Cytotoxicity of Bark Extracts and Organic Compounds

In order to determine the cytotoxicity of the bark extract or organic compound, 200 μl of each cell (each cell used in Experimental Examples 2 to 4) was added to each well of a 96-well plate at a concentration of 5 × 10 ⁴ cells / mL. C was incubated for 1 day in a CO ₂ incubator. In this case, DMEM medium containing 10% of FBS and 1% of antibiotics was used. After incubation, the extract was added for each concentration, followed by incubation for 24 hours under the same culture conditions. After incubation, 5 mg / mL MTT (Thiazolyl Blue Tetrazolium Bromide) reagent was added and incubated for 3 hours under the same conditions. Thereafter, the culture medium was removed, 100 μl of DMSO (Dimetyl sulfoxide) was added thereto, followed by stirring for 10 minutes. The resultant was measured at 570 nm using a micro plate reader. Calculated.

According to the results shown in the line graph of Figures 4 to 14 it was confirmed that there is no toxicity at all extract concentrations used in the experiment.

Hereinafter, preferred and various cosmetic formulations for use in the present invention are disclosed, but the scope of the present invention is not limited to the disclosed formulations.

[Manufacture example 1] lotion

The lotion containing the present invention bark tree extract obtained in Examples 1 to 3 was prepared as shown in Table 6 below.

Raw material Raw material Content (% by weight) One The invention of the bark tree extract 10.0 2 glycerin 3.0 3 Butylene Glycol 2.0 4 Propylene glycol 2.0 5 Polyoxyethylene Cured Castor Oil 1.0 6 ethanol 10.0 7 Triethanolamine 0.1 8 antiseptic a very small amount 9 Pigment a very small amount 10 Spices a very small amount 11 Purified water Remaining amount

[Preparation Example 2] Lotion The present invention The lotion containing the present invention bark extract obtained in Examples 1 to 3 was prepared as shown in Table 7.

Raw material Raw material Content (% by weight) One The invention of the bark tree extract 10.0 2 Beeswax 1.0 3 Polysorbate 60 1.5 4 Solvidan Sesquioleate 0.5 5 Floating paraffin 10.0 6 Sorbidate stearate 1.0 7 Lipophilic Monostearine Glycerin 0.5 8 Stearic acid 1.5 9 Glyceryl Stearate / Pig-400 Stearate 1.0 10 Propylene glycol 3.0 11 Carboxy Polymer 0.1 12 Triethanolamine 0.2 13 antiseptic a very small amount 14 Pigment a very small amount 15 Spices a very small amount 16 Purified water Remaining amount

Preparation Example 3 Cream of the Invention The cream containing the present invention bark tree extract obtained in Examples 1 to 3 was prepared as shown in Table 8 below.

content Raw material Content (% by weight) One The invention of the bark tree extract 10.0 2 Stearic acid 2.0 3 Cetyl alcohol 2.0 4 Glyceryl Monostearate 2.0 5 Polyoxyethylene sorbidan monostearate 0.5 6 Solvanesesquioleate 0.5 7 Glyceryl Monostearate / Glyceryl Stearate / Polyoxyethylene Stearate 1.0 8 Wax 1.0 9 Floating paraffin 4.0 10 Squalane 4.0 11 Caprylic / Capric Triglycerides 4.0 12 Carboxy Vinyl Polymer 0.3 13 Butylene glycol 5.0 14 glycerin 3.0 15 Triethanolamine 0.5 16 Purified water Remaining amount

Preparation Example 4 Preparation of Essence The Essence containing the present invention bark tree extract obtained in Examples 1 to 3 was prepared as shown in Table 9 below.

content Raw material Content (% by weight) One The invention of the bark tree extract 10.0 2 Cytostolol 1.7 3 Polyglyceryl 2-oleate 1.5 4 Ceramide 0.7 5 Steares-4 1.2 6 cholesterol 1.5 7 Dicetylphosphate 0.4 8 Concentrated glycerin 5.0 9 Macadamia oil 15.0 10 Carboxy Vinyl Polymer 0.2 11 Xanthan Gum 0.2 12 antiseptic a very small amount 13 Spices a very small amount 14 Purified water Remaining amount

From the above description, those skilled in the art will understand that the present application can be implemented in other specific forms without changing the technical spirit or essential features. In this regard, it should be understood that the embodiments described above are exemplary in all respects and not restrictive. The scope of the present application should be construed that all changes or modifications derived from the meaning and scope of the following claims and equivalent concepts rather than the detailed description are included in the scope of the present application.

The present application is a very useful invention in the natural cosmetics industry because it has the effect of providing a skin anti-inflammatory, anti-wrinkle and whitening functional cosmetic composition containing the extract of the bark or its fraction as an active ingredient.

Claims (2)

Anti-inflammatory and anti-wrinkle functional cosmetic composition containing any one or more of the organic compounds of Formula 1 to Formula 5 in the following chemistry.
[Formula 1]

… [Formula 1]
[Formula 2]

… [Formula 2]
[Formula 3]

… [Formula 3]
[Formula 4]

… [Formula 4]
[Formula 5]

… [Formula 5] According to claim 1, wherein the cosmetic composition is a cosmetic composition, characterized in that the formulation of any one selected from lotion, lotion, cream and essence.

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