KR20190003010A - A cosmetic composition containing cold water extract or its ethyl acetate fraction from Sanguisorba officinalis having specific anti-microbial activity against acne-inducing bacterium - Google Patents

Abstract

The present invention relates to a cosmetic composition containing a cold water extract of Sanguisorba officinalis L. or an ethyl acetate fraction thereof, wherein the cold water extract of Sanguisorba officinalis L. is obtained by extracting, with cold water, roots (sanguisorba roots) of Sanguisorba officinalis L. having specific anti-microbial activity against Propionibacterium acnes. The cosmetic composition is capable of preventing or improving acne and can be manufactured into compositions for cosmetics, soap, or cleansing agents.

Description

[0001] The present invention relates to a cosmetic composition comprising an oily cold extract having a specific antibacterial activity against bacteria causing acne, or a fraction thereof. [0002]

The present invention relates to a method for producing an extract of Sanguisorba officinalis L. by extracting the root portion of Sanguisorba officinalis L. having specific antibacterial activity with Propionibacterium acnes with cold water and then extracting the extract or its ethyl acetate fraction ≪ / RTI >

Acne vulgaris is the most common inflammatory disease of the pilosebaceous unit of the skin, most commonly on the face of adolescent men, and occasionally on the back, back, chest or shoulder. The main cause is the effect of the male hormone (androgen) which induces the production of excess sebum along with the expansion of sebaceous glands and the formation of comedones. Another major cause is the erosion of abnormal hair follicle epithelium (skin keratin) and the proliferation of propionibacterium acnes.

Propionibacterium acnes are anaerobic, gram positive, bacillus, which live as symbiotic bacteria in the cleavage site. The exfoliation of abnormal hair follicle epithelium induces anaerobic conditions in the hair follicle and promotes the proliferation of propionibacterium acnes. In addition, Propionibacterium acnes secretes lipase and a chemotactic factor and metabolizes glycerol component of excessively secreted sebaceous triglyceride into free fatty acids. These extra free fatty acids promote the production of inflammatory leukocytes around the hair follicles. On the other hand, inhibition of the proliferation of propionibacterium acnes can minimize rupture of the cotton buds and inhibit the development of acne.

Several oral or transdermal antibiotics have been used to treat acne, such as tetracycline, erythromycin, and clindamycin. Prolonged use of these broad spectrum antibiotics can cause overgrowth of Candida albicans and resistance to antibiotics. In addition, transdermal chemical agents such as benzoyl peroxide, azelaic acid and retinoic acid, known as anti-acne treatments, have side effects such as skin irritation and contact dermatitis.

Recently, many antimicrobial plants have been reported internationally in order to solve the above problems. Some of these can be used as an alternative to acne treatment. The vast majority of extracts which showed pronounced antimicrobial activity against propionibacterium acnes in extracts obtained from the target tissues of presently reported single plants, including plants, are methanol (patent documents 0001, 0002, 0003 and non-patent documents 0001, 0002 , 0003, 0004), ethanol (Patent Document 0004, 0005, 0006, 0007, and Non-Patent Document 0005, 0006, 0007, 0009, 0010, 0011), hot water (Patent Document 0009, ), Phosphoric acid buffer solution (Patent Document 0011), acetone (non-patent document 0013), other organic solvents (Non-Patent Document 0014), and steam distillation method (Patent Document 0012 and Non-Patent Document 0016).

The 'Halal' market is one of the largest markets in the international market. It is also a growing market. It is a Muslim lifestyle and a halal market. In the case of cosmetics, in order to satisfy the halal standard, ethanol widely used for extracting and concentrating an active ingredient in a plant can not be used at all, and polyhydric alcohols can not also be used as an animal-derived ingredient.

Also, the strains used in the fermentation process should be selected according to strict criteria and ethanol produced during the fermentation process may also be an obstacle. Therefore, the only alternative is the extraction method using cold water and hot water, which has the advantage of easily extracting the active ingredient compared with other extraction methods.

The perennial plant, S. officinalis L., belonging to Rosacease, is widely distributed in the Far East region including Korea. The roots of Oleaceae have been used as herbal medicines in oriental medicine (Non-Patent Document 0017).

Jiwoo has been mainly used in the treatment of hemostatic and wound areas in oriental medicine, and is known to externally use for dermatitis, mucositis, eczema, burns (Non-Patent Document 0018). It contains ziguglycoside I, II and pomolic acid. Branches contain quercetin and kaempferol glycosides and triterpenoid saponin such as ursolic acid. Vitamin C on leaf, pharyngeal component such as chrysanthemin and cyanin on flower (Non-Patent Document 0019).

Based on the above-mentioned pharmacological components, there have been reported studies on immunosuppressive activity, antimutagenic effect, nerve cell damaging inhibitory effect, antioxidative effect, hypersensitivity allergy prevention, anticancer, antibacterial effect, and hemostatic action from Oyster (Non-Patent Document , 0020, 0021, 0022, 0024, 0024, 0026, 002, 0027).

Patents for extracts of oat extracts include Streptococcus mutans-inhibiting oral detergent compositions (Patent Document 0013) that cause cavities, antiviral active compositions for VHSV and IHNV causing septicemia (Patent Document 0014), skin code effect cosmetic compositions , A composition for prevention and improvement of diabetes (Patent Document 0016), a composition for prevention and improvement of listeria or salmonella-induced sepsis (Patent Document 0017), and the like.

Korean Patent Publication No. 10-2015-0000723, 2015.01.05. open Korean Registered Patent No. 10-1507889, Apr. 20, 2015. Announcement Korean Patent Publication No. 10-2008-0044948, May 22, 2008. open Korean Patent Publication No. 2002-0003313, 2002.01.12. open Korean Patent Publication No. 2000-0019981, Apr. 15, 2000. open Korean Patent Publication No. 10-2012-0086123, Aug. 22, 2012. open Korean Patent Laid-Open Publication No. 10-2014-00992233, 2014.08.11. open Korean Patent Publication No. 10-2004-0081533, September 22, 2004. open Korean Patent Publication No. 10-0700911, March 28, 2007. Announcement Korean Registered Patent No. 10-0700912, March 28, 2007. Announcement Korean Patent Publication No. 10-2007-0110589, 2007.11.20. open Korean Patent Publication No. 2003-0016818, 2003.03.03. open Korean Patent Publication No. 10-2013-0113018, Oct. 15, 2013. open Korean Patent Publication No. 10-2016-0150177, December 29, 2016. open Korean Patent Publication No. 10-2009-0117199, November 12, 2009. open Korean Patent Publication No. 10-2016-0102638, Aug. 31, 2016. open Korean Patent Publication No. 10-2017-0055104, Jul. open

 Muddathir et al., J. Woos Sci. 2013, Vol. 59, pp. 426-431  Kim et al., J. Microbial. Biotechnol. 2010, Vol. 20, pp. 82-87  Lim et al., J. Microbiol. 2007, Vol. 45, pp. 473-477  Tsai et al., Food Chem. 2010, Vol. 119, pp. 964-968  Tangje Arung et al., Biodiversitas 2017, Vol. 18, pp. 321-325  Pothitirat et al., Pharm. Biol. 2010, Vol. 48, pp. 182-186  Sharma and Lall, S. Afr. J. Sci. 2014, Vol. 110, Aer. # 2013-0293  Sharma et al., Phytother. Res. 2011, Vol. 25, pp. 517-521  Chomnawang et al., Fitoterapia 2007, Vol. 78, pp. 401-408  Wang et al., J. Agric. Food Chem. 2013, Vol. 61, pp. 12274-12282  Kumar et al., Trop. J. Pharm. Res. 2007, Vol. 6, pp. 717-723  Sathishkumar et al., J. Photocheml. Photo Biol., B: Biology 2010, Vol. 20, pp. 82-87  Vijayalakshimi et al., Int. J. PharmTech Res. 2011, Vol. 3, pp. 320-327  Shah et al., J. Med. Plants Res. 2015, Vol. 9, pp. 755-763  Pothitirat et al., Med. Princ. Pracy. 2010, Vol. 19, pp. 281-286  Qidwai et al., Int. J. Pharm. Sci. Res. 2016, Vol. 7, pp. 2917-2924  An et al., Kor. J. Kor. Soc. Appl. Biol. Chem. 2004, Vol. 47, pp. 244-250  Son et al., J. Kor. Acad. Oral Health 2015, Vol. 28, pp. 97-104  Lee et al., J. Korean Soc. Food Sci. Nutr. 2005, Vol. 34, pp. 1545-1552  An et al., Kor. J. Kor. Soc. Appl. Biol. Chem. 2004, Vol. 47, pp. 131-145  Ban et al., Kor. J. Med. Crop Sci. 2005, Vol. 13, pp. 219-226  Goun et al., J. Ethnopharm. 2002, Vol. 81, pp. 337-342  Hwang et al., J. Physiol. Pathol. Kor. Med. 2014, Vol. 28, pp. 499-505  Kim et al., Nat. Prod. Sci. 2002, Vol. 8, pp. 177-182  Kim et al., Kor. J. Food Cook. Sci. 2011, Vol. 27, pp. 15-26  Park et al., Kor. J. Herb. 2012, Vol. 27, pp. 23-29  Park et al., J. Food Preserv. 2012, Vol. 19, pp 744-750

It is an object of the present invention to provide a cosmetic composition comprising a cold water extract of a castor oil (root of oat) and an ethyl acetate fraction thereof having a specific antibacterial activity against Propionibacterium acnes.

In order to achieve the above object, the present invention provides a method for preventing or ameliorating acne, which comprises a cold water extract of root oil (root of oat) or its ethyl acetate fraction having propionic acid, The present invention provides a cosmetic composition.

As described above, according to the present invention, the cold water extract or the fraction thereof of the crude oil has an antibacterial activity specific to Propionibacterium acnes, and thus is effective for prevention or improvement of acne. Accordingly, the composition can be used as a cosmetic composition, more specifically, a cosmetic composition, a soap and a detergent composition.

1 is a flow chart of a process of separating active fractions from cold water extract (CWE) of crude oil.
2 is a paper disc agar diffusion method in which antimicrobial activity of CWE and ethyl acetate fraction (CWE-EA) of cold water extract against Propionibacterium acnes is analyzed. Figure (A) is a photograph of inhibitory rings against acne bacteria in CWE 10 mg / disk (1), untreated (2), and distilled water (3) Figure (B) is a photograph of the inhibitory ring against acne bacteria in the distilled water treatment (1) and CWE 1 mg / disk treatment (2), 3 mg / disk treatment (3) and 5 mg / . Figure (C) shows the inhibitory effect on the acne bacterium in the distilled water treatment (1) and CWE-EA 1 mg / disk treatment (2), 3 mg / disk treatment (3) and 5 mg / It is a photograph.
FIG. 3 is a graph showing the results of a paper disc agar diffusion method using n-hexane, ethyl acetate, n-butanol and water fractions of cold water extract (CWE) The results are compared with the inhibition of acne by.
FIG. 4 shows the results of confirming whether colonies were formed on a solid medium to confirm the minimum inhibitory concentration (MIC) and the minimum bactericidal concentration (MBC) of the cold water extract (CWE) of crude oil on Propionibacterium acnes.
FIG. 5 is a graph showing the results obtained by pouring the ethyl acetate fraction of cold water extract (CWE) of crude oil onto a silica gel 60 glass plate and using a mixed solvent of ethyl acetate, n-hexane, chloroform, acetic acid (40: 30: 30: / v / v / v) and analyzed by UV wavelength 254 nm (A), 366 nm (B) and sulfuric acid spray coloring method (C).
FIG. 6 shows the ethyl acetate fraction of the cold water extract (CWE) of crude oil after being developed on TLC and analyzed by HPLC. (2) caffeic acid, (3) ferulic acid, (4) quercetin, and (5) kaempferol.

One embodiment of the present invention can provide a cosmetic composition for improving acne comprising cold water extract of Sanguisorba officinalis.

Oylum is a perennial plant belonging to Rosaceae. It is distributed in Korea, Japan, Europe and Manchu China. After picking the leaves, the smell of cucumbers is called oyula. In one room, the roots of Oylus are used as a medicinal herb. It contains 17% tannin, 2.5% saponin and Sanguisorbigenin, and the stem and leaf contain qurcetin, Kaemferol glycoside and Ursolic acid. Jili (Root of Oleaceae) in one room to lower the heat of the body and has the effect of stopping bleeding, is known to have detoxification. In the case of castor oil (roots of Oleacea), it is effective for burn treatment, hemostatic action, vomiting suppression action, antibacterial action and the like.

The cold water extract of the ores may be extracted at 1 to 20 ° C for 12 to 36 hours, preferably at 2 to 15 ° C for 18 to 30 hours, most preferably at 3 to 10 ° C For 24 hours.

In the present invention, the cosmetic composition may be characterized by inhibiting Propionibacterium acnes, which is an acne bacterium, and most preferably KCTC3314.

In the present invention, the Sanguisorba officinalis may be at least one kind of region extract selected from the group consisting of flowers, leaves, stalks, fruits, seeds, roots and toppings of Oleaceae, It can be a root.

In the present invention, the cosmetic composition may be a solution, a suspension, an emulsion, a paste, a gel, a cream, a lotion, a powder, a soap, a surfactant-containing cleansing oil, a powder foundation, an emulsion foundation, a wax foundation, , And may be a soap or a surfactant-containing cleansing agent.

In the present invention, the cold water extract of Sanguisorba officinalis may be contained at a concentration of 1000 μg / mL to 1500 μg / mL, preferably 1250 μg / mL.

In the present invention, it may be characterized by comprising an ethyl acetate fraction obtained by fractionating the cold water extract of Sanguisorba officinalis with ethyl acetate.

In the present invention, the ethyl acetate fraction may include any one or more active ingredients selected from the group consisting of hydroxycinnamic acids and flavonols.

In the present invention, the ethyl acetate fraction may be contained at a concentration of 100 μg / mL to 200 μg / mL, preferably 156 μg / mL.

Hereinafter, the present invention will be described in more detail with reference to examples, but the present invention is not limited to the following examples.

Cold water extract and fraction

1. Production of cold water extract (CWE)

Oylum root was washed with distilled water, dried and crushed finely with a grinder. 10 g of water at 4 ° C was added to the pulverized sample (50 g), and the mixture was extracted for 24 hours. The filtrate was filtered through Wattmann paper filter paper (Whatman No. 2), and the filtrate was concentrated under reduced pressure at 65 ± 2 ° C. And then lyophilized with a freeze dryer to obtain 0.2916 g of the final extract. The yields from cold-water extraction from cold storage are shown in Table 1 below.

Yield from cold water extraction from castor oil Building Freeze-dried yield of cold water extract (CWE) 50 g 0.2916 g

2. Preparation of fraction of cold water extract (CWE fraction)

After a certain amount of water was added to the cold water extract, each fraction was extracted by sequentially using n-hexane, ethyl acetate, butanol, and water as extraction solvents having different polarities using a separating funnel. First, n-hexane was added to the separatory funnel in the same amount as distilled water, mixed well, and left to separate into two layers, an aqueous layer and an extraction solvent. The extraction solvent was separated and the remaining filtrate was again extracted with ethyl acetate and butanol in the same manner. At this time, the extraction solvent was added in the same amount as the remaining filtrate. A schematic diagram of the extraction and fractionation process is shown in Fig. As a result, the sequential fractions were prepared by suspending 50 g of the cold water extract of the crude oil in 500 mL of distilled water and then sequentially fractionating the mixture with n-hexane (500 mL), ethyl acetate (500 mL), n-butanol ≪ / RTI >

Antimicrobial activity of plant extracts and fractions

The antimicrobial activity of Gram-negative bacteria (2 species) and Gram-positive bacteria (3 species) were measured using paper disk diffusion method, minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) Respectively.

1. Target strains and culture conditions

The Gram-positive bacteria Gram positive bacterium, Propionibacterium acnes (KCTC3314), were cultured in a brain-heart infusion (BHI) medium for 2 days or more under anaerobic conditions, and Listeria monocytogenes Staphylococcus aureus was cultured on BHI medium under aerobic conditions. The Gram-negative bacteria Escherichia coli and Vibrio parahaemolyticus used in the present invention were cultured in Luria-Bertani (LB) medium. Each strain was incubated at 37 ° C and 200 rpm for 1 to 2 days with shaking. Growth of each strain and lysis of bacteria were measured by a Biochrom Libra S22 spectrophotometer. Lt; / RTI >

2. Paper disc agar diffusion method

The cultured strains were uniformly plated on a BHI agar plate or LB agar plate using a sterile cotton swab at a constant concentration (optical density at 600 nm, absorbance value 0.5). After drying the plates, paper discs (Φ 6 mm, Adventec. USA) were placed on the plates inoculated with the strains and each of the cold water extracts was absorbed at 1, 3, 5 or 10 mg / disc. The plate treated with the sample was incubated at 37 ° C for 48 hours, and the antimicrobial activity was measured to the extent of the inhibition zone around the disk. At this time, antibiotics (ampicillin, kanamycin, spectinomycin, timentin) were used as a control group and the treatment concentration was 20 g / disc. As a result, strong anti-acne activity was observed at 10 mg / disk of cold water extract of crude oil (Fig. 2A and Table 2). And excellent anti-acne activity was observed up to 1 mg / disk treatment even at the concentration-dependent treatment (Fig. 2B). However, no antibacterial activity was observed for the other four bacteria. These results indicate that the cold water extract of Jiu-Yi has a specific antimicrobial activity against Propionibacterium acnes, but has no antibacterial activity against other bacteria.

 Antibacterial activity against Gram positive bacterium, Propionibacterium acnes Samples (10 mg / disc) Growth inhibition ring size (cm) Cold Water Extract (CWE) 3.2 x 3.0 ampicillin - kanamycin 3.1 x 3.0 spectinomycin - timentin -

In addition, excellent anti-acne activity was observed in the ethyl acetate fraction obtained from the cold water extract of the crude oil, but it was weak or not observed in the hexane, butanol and distilled water fractions (Fig. 3). As a result of treatment with the concentration of ethyl acetate fraction, excellent anti-acne activity was confirmed up to 1 mg / disk treatment (Fig. 2C).

3. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of cold water extract (CWE) and ethylacetate fraction (CWE-EA)

Propionibacterium acnes were cultured in a test tube containing BHI broth medium for 24 hours at 37 ° C. Here, the cold water extract and the ethylacetate fraction were diluted by two-fold dilution and added to the test tube. MIC was determined as the minimum concentration that inhibited the visible growth of the bacteria after incubation for 48 hours in a test tube inoculated with a bacterial dilution and a sample. As shown in Table 3 below, the MIC values of the cold water extract and ethylacetate fractions against propionibacterium acnes were determined to be 1250 and 156 μg / mL. The number of colonies observed on the plate was counted after culturing the bacteria on a solid medium of 100 μL of the culture medium of the test tube corresponding to a concentration higher than the concentration selected by MIC. MBC was used to determine the concentration of the extract to be treated at which 100% of the colonies were confirmed to be killed, and the concentration was determined as shown in [Table 3] and [Figure 4]. As shown in Table 3 below, the MBC values of the cold water extract and ethylacetate fractions against propionibacterium acnes were determined to be 2500 μg / mL and 312 μg / mL.

 The minimum inhibitory concentration and minimum germicidal concentration of the ethyl acetate fraction of the crude oil-extracted cold extract and the extract against the propionibacterium acnes extract MIC ([mu] g / mL) MBC (占 퐂 / mL) Cold Water Extract (CWE) 1250 2500 The ethyl acetate fraction (CWE-EA) 156 312

Analysis of Components of Ethyl Acetate Fractions Having Antimicrobial Activity

1. Thin layer chromatography (TLC)

The ethyl acetate fraction with antimicrobial activity was extracted with ethyl acetate / hexane / chloroform / acetic acid (40: 30: 30: 2, v / v) using a silica gel 60 glass plate (20 x 20 cm, Merck, DarDarmstadt, Germany) / v / v / v) mixed solvent, and the ratio of ethyl acetate fraction and standard substances (caffeic acid, ferulic acid, quercetin, kaempferol) was measured by UV lamp (254 nm and 366 nm wavelength) Rf values were compared and analyzed. As a result of the above experiment, it was confirmed that bands similar to the Rf values of caffeic acid, ferulic acid, quercetin and kaempferol were observed [Fig. 5.]

2. High performance liquid chromatography (HPLC)

The ethyl acetate fraction was dissolved in methanol, filtered (0.45 μm filter) and loaded on a ZORBAX Eclipse Plus C18 column (4.6 × 150 mm, 5 μm, Agilent Technologies, Lawrence, KS) using HPLC (1260 Infinity Quaternary LCSystem, Agilent Technologies, Waldbronn, Germany) USA) at 254 nm. Injection volume was 10 μL and the mobile phase was solvent A (2% acetic acid in 30% methanol) and B (2% acetic acid in 50% acetonitrile) at a flow rate of 1 mL / / v) to 70 minutes at a ratio of 0: 100 (v / v). The content was determined by the regression equation of the calibration curve prepared using the product caffeic acid, ferulic acid, quercetin and kaempferol. As a result of the above tests, two kinds of hydroxycinnamic acids (caffeic acid, ferulic acid) and two kinds of flavonols (quercetin and kaempferol) were identified by the reversed phase HPLC method by comparing the retention time with the standard substance in the fractions of Cheongmaeae 6]. Among the ethylacetate fractions of the crude oil extracts, various unknown substances including caffeic acid, ferulic acid, quercetin and kaempferol were detected, and ferulic acid content was the highest.

Claims (9)

A cosmetic composition for improving acne comprising cold water extract of Sanguisorba officinalis. The cosmetic composition for improving acne according to claim 1, wherein the cosmetic composition inhibits Propionibacterium acnes, an acne bacterium. [Claim 7] The cosmetic composition for improving acne according to claim 1, wherein the Sanguisorba officinalis is at least one region extract selected from the group consisting of flowers, leaves, stems, fruits, seeds, roots and outposts of Olefins. The cosmetic composition of claim 1, wherein the cosmetic composition comprises a solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, surfactant containing cleansing, oil, powder foundation, emulsion foundation, wax foundation, Wherein the cosmetic composition has at least one formulation selected from the group consisting of: The cosmetic composition for improving acne according to claim 1, wherein the cold water extract of Sanguisorba officinalis is contained at a concentration of 1000 μg / mL to 1500 μg / mL.  The cosmetic composition for improving acne according to claim 1, which comprises an ethyl acetate fraction obtained by fractionating the cold water extract of Sanguisorba officinalis with ethyl acetate. 7. The cosmetic composition for improving acne according to claim 6, wherein the ethyl acetate fraction comprises at least one active ingredient selected from the group consisting of hydroxycinnamic acids and flavonols. 7. The cosmetic composition for improving acne according to claim 6, wherein the ethyl acetate fraction is contained at a concentration of 100 mu g / mL to 200 mu g / mL. Quasi-drugs including cold water extracts of Sanguisorba officinalis or fractions thereof.

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