KR102282642B1 - Cosmetic composition containing the extract of 5 aromatic herbs and 5 medicinal herb - Google Patents

Abstract

The present invention relates to a cosmetic composition for improving a skin condition with an excellent anti-wrinkle effect, which comprises a complex extract of five aromatic herbs of Boswelliacarterii, Agastacherugosa, Inulahelenium, Clove, Eugenia caryophyllus and Sandalwood, Santalumalbum, and five medicinal herbs of Panaxginseng, Glycyrrhizauralensis, Cinnamomumcassia, Poriacocos and Asparagus cochinchinensis, thereby providing antibiosis against skin lesion-causing bacteria, skin damage recovery and regeneration by promoting keratinocyte migration, whitening and collagen production by inhibiting melanin production.

Description

Cosmetic composition containing the extract of 5 aromatic herbs and 5 medicinal herbs

The present invention relates to a cosmetic composition containing a complex extract of five aromatic plants and five herbal materials as active ingredients, specifically, aromatic plants Boswellia carterii , Agastache rugosa , Inula helenium ), clove, Eugenia caryophyllus , Sandalwood, Santalum album , ginseng ( Panax ginseng ), licorice ( Glycyrrhiza uralensis ), cinnamon tree ( Cinnamomum cassia ), Poria cocos , and cochinchinensis Asparagus ) It relates to a cosmetic composition containing a complex extract as an active ingredient, which has excellent antibacterial effect on skin lesion-causing bacteria, keratinocyte migration promoting effect, melanin production inhibition and collagen production effect.

Following the novel influenza and Middle East Respiratory Syndrome, various infectious diseases have recently become prevalent, including the novel coronavirus infection, and various skin problems have occurred due to these infectious diseases changing lifestyles such as wearing personal hygiene tools for a long time, reducing outdoor activities, and frequent use of mobile phones. . In particular, Staphylococcus aureus and Cutibacterium acnes are representative bacteria that cause such skin troubles. They coexist and interact in high-temperature and high-humidity environments such as when wearing a mask, causing skin diseases such as acne.

S. aureus is the bacterium that causes most skin infections, increases in hot and humid environments, and is a major cause of bacterial skin diseases and acne caused by personal hygiene (Nuo-Ya Zhou et al ., 2020). In addition, as the use of smartphones increases because outside activities are not free, the short-wavelength visible light of smartphones induces the proliferation of S. aureus and causes acne (M. Singh et al ., 2020). Vitamin D produced by sunlight increases skin immunity and increases antibacterial activity against S. aureus (Jeffrey W. Wang et al ., 2015). However, it is difficult to fully expect skin immunity against S. aureus now that external access is not as active as in the past for daily quarantine.

Although C. acnes is a flora that accounts for half of the skin flora, it is a bacterium that causes acne when the skin condition is deteriorated (Chia-Jung Lee et al ., 2017). According to a recent report, quantitative changes in C. acnes were not observed in acne skin, but biofilms secreted by these bacteria are known to cause inflammation and antibacterial resistance, thereby affecting acne exacerbation (Eftychia Platsidaki and Clio Dessinioti, 2018). C. acnes also interacts in S. aureus skin and thereby deteriorate the skin lesions caused by S. aureus (Chih-Wei Lo et al., 2011).

Wearing personal hygiene tools for a long time causes various troubles and damage to the skin. Acne caused by wearing a mask deposits pigment in the wound area, and the sunlight reflected by the mask stimulates the production of melanin around the eyes and forehead, which can make the skin dull. Also, personal hygiene tools attached to the face cause physical scars on the skin and deep wrinkles.

Acne, skin damage, hyperpigmentation of the skin, and wrinkle formation cause skin troubles for many people not only in infectious disease situations but also in everyday life. Therefore, there is a need to develop a new material that is safer for the human body while exhibiting the efficacy of improving such skin problems.

The present inventors have been searching for natural materials exhibiting this effect, and the complex extracts of aromatic plants such as frankincense, baechohyang, mulberry, clove and dandelion and herbal materials such as ginseng, licorice, broiler, bokryeong and cheonmundong have been used for S. The present invention was completed by confirming that it is effective in improving the skin condition due to its excellent antibacterial effect, biofilm formation inhibitory effect of C. acnes, keratinocyte migration promoting effect, melanin production inhibition and collagen production effect.

(0001) Republic of Korea Patent No. 10-2184758 (2020.11.24) (0002) Republic of Korea Patent Publication No. 10-2018-0043580 (2018.04.30.) (0003) Republic of Korea Patent Registration No. 10-2199858 (2021.01.04) (0004) Republic of Korea Patent Registration No. 10-2010404 (2019.08.07)

The present invention aims to provide a cosmetic composition for improving skin condition containing, as an active ingredient, a complex extract of aromatic plants, frankincense, baechohyang, woodhyang, clove, and dandelion, and herbal materials such as ginseng, licorice, broiler, bokryeong and cheonmundong. .

According to the present invention to achieve the above object, there is provided a cosmetic composition containing frankincense, baechohyang, woodhyang, clove, danhyang, ginseng, licorice, broiler, bokryeong and cheonmundong complex extract as an active ingredient.

The complex extracts are frankincense, baechohyang, mulberry, clove, dandelion, ginseng, licorice, broiler, bokryeong and cheonmundong 1~2:1~2:1~2:1~2:1~2:1~2:1, respectively. It is prepared by extraction by mixing in a weight ratio of ~2:1~2:1~2:1~2.

The complex extract as an active ingredient is contained in an amount of 0.001 to 10% by weight based on the total weight of the cosmetic composition.

The cosmetic composition is characterized in that it is for antibacterial. The cosmetic composition is characterized in that it is for skin regeneration. The cosmetic composition is characterized in that for skin whitening. The cosmetic composition is characterized in that it is for improving skin wrinkles.

The cosmetic composition containing frankincense, baechohyang, woodhyang, clove, danhyang, ginseng, licorice, broiler, bokryeong and cheonmundong complex extract according to the present invention as an active ingredient contains a natural extract and is safe and has excellent antibacterial effect due to its synergistic action , promotes skin regeneration and skin damage recovery, skin whitening and wrinkle improvement.

Hereinafter, the contents of the present invention will be described in detail to help the understanding of the present invention.

According to the present invention, there is provided a cosmetic composition containing, as active ingredients, a complex extract of five aromatic plants, frankincense, baechohyang, mulberry, clove and dandelion, and five herbal ingredients, ginseng, licorice, broiler, bokryeong and cheonmundong.

Representative aromatic plants that emit fragrance from plants include Pineaceae, Camphoraceae, Rusaceae, Cloveaceae, Umbellaceae, Lamiaceae, Asteraceae, Asteraceae, Ashaceae, and Honeysuckle. After the Goryeo Dynasty, our ancestors began to actively use aromatic plants in real life. , bathing, insect repellent, and perfumery (Minji Hong et al., 2020).

Frankincense is a dwarf shrub belonging to the olivine family. When the stem is cut, milky sap flows out and hardens into aromatic resin. This is called frankincense and has been used as a material for perfumes and cosmetics since ancient times. It is also widely used for various therapeutic purposes. Frankincense is known for its analgesic and anti-inflammatory effects, and recently it has been reported to be effective for stress and mental treatment. In addition, frankincense is non-toxic and has antiseptic and strong bacterial growth inhibitory effects (Park Jeong-suk, 2017).

Baechohyang is a perennial herb belonging to the family Lamiaceae and is a representative Korean aromatic plant. Due to the strong scent of baecho, it is used as a raw material for various industrial products such as food and spices. Also called Kwakhyang (藿香) in oriental medicine, it has been widely used as Kwakhyangjeonggi-tang for stomach pain accompanied by food-induced congestion, chills, and diarrhea. Anti-inflammatory, antioxidant, anti-anxiety, etc. are known as physiological activities of baechohyang extract, and it has been reported that the essential oil of baechohyang has antibacterial, antifungal, antiviral, and insecticidal activities (Minji Hong et al., 2020).

Mokhyang is a perennial herb belonging to the Asteraceae family, and there have been reports of anticancer, anti-obesity, osteoclast differentiation, anti-inflammatory, and antidiabetic effects in studies on Mokhyang (Oh, Jaesun et al., 2014). In addition, it is known that sesquiterpene and sesquiterpene lactone-based compounds, which are major components of mulberry, exhibit antiviral, antibacterial, anti-inflammatory, and cancer cell proliferation inhibitory effects (Ayoung Lee et al., 2007).

Clove is an evergreen small arboreous tree belonging to the family Clove, especially when the flower buds turn from green to red, the flowers are harvested and dried, called cloves. Cloves have a strong distinctive odor and strong taste, and have been used in Korea for a long time to enhance flavor, suppress bad flavor, etc., and are used for insect repellent and mold control. The physiological activity of cloves has been reported to have an effect on leukemia cell differentiation and an allergic reaction suppression effect, and antioxidant, antibacterial, and antiviral effects are known (Jinman Lee and Sangseon Huh, 2014).

Danhyang (檀香) is an evergreen tree belonging to the Danhyang family, also called sandalwood. Danhyang has a sweet and sweet fragrance and is widely used as an air freshener because of its excellent durability. Also, since ancient times, it has been considered sacred as a material for religious events, such as incense and Buddha images, and has been used as a raw material for various crafts. The physiological activity of Danhyang is known for its antiviral, anticancer, analgesic, and antioxidant effects (Kim Tae-Hoon, 2008).

Herbal materials have been used for preventing, diagnosing, improving and treating various diseases since ancient times based on their excellent efficacy, and they help users to maintain their health and improve their quality of life. Through recent research, various physiological activities possessed by herbal medicines as well as previously known efficacy are scientifically verified, and a number of studies are being conducted on the efficacy of improving skin condition.

Ginseng (人蔘) is a perennial herb belonging to the Orchidaceae family, and its root is used as an important medicinal agent, and it is prescribed together with various medicinal substances and is one of the most important herbal medicines used from ancient times to the present as a treatment for numerous ailments (Lee Sang-myung, 2014). The main physiological activities of ginseng include anti-stress, immune function enhancement, vitality enhancement, and anti-cancer effects, and skin-related effects are known for anti-wrinkle, skin aging inhibition, and whitening effects (Mijeong Lee et al., 2008; Hyeonjeong Lee et al. , 2010).

Licorice (甘草) is a perennial herb belonging to the legume family, and the dried root is used as a medicine. Bangyakhappyeon, the most commonly used oriental medicine prescription book in Korea, is the herbal medicine with the highest frequency of prescription among 293 types of water rejuvenating drugs (Lee Sang-myung, 2014). Licorice is known to have neutralizing and detoxifying effects, and in relation to the skin, the effects of promoting collagen production, whitening, anti-inflammatory, anti-acne, antioxidant, and wound healing have been reported (Jinyoung Lee et al ., 2012; Baocheng Hao et al. , 2020).

The broiler tree (肉桂) is an evergreen tree belonging to the camphor family and is mainly used as a herbal medicine for people with weak constitution and poor blood flow. It is also known to have effects such as lowering blood pressure, antispasmodic action, raising blood sugar, anti-allergy, and improving gastrointestinal diseases (Kwan-Ha Park et al., 2001; Jae-Yun Cha et al., 2018). Recently, excellent antioxidant, anti-inflammatory, antihistamine, antibacterial and whitening effects of broilers have been reported, and the potential as a natural functional cosmetic material has been confirmed (Yeong-Sook Lee and Min-Jeong Yoo, 2019).

Bokryeong is a basidiomycete mushroom belonging to the family Fungi family. It parasitizes on the roots of conifers, especially pines, and grows by agglomeration of mycelium in the ground. Since ancient times, bokryeong is one of the herbal medicines that has been widely used in oriental medicine as it has excellent effects in relieving chronic gastritis, diuretic action, immunity enhancement, appetite enhancement, blood sugar lowering, mental stability and heart contraction (Son Miye, 2007). In addition, bokryeong is effective against skin cancer and is known to exhibit anti-oxidation, anti-wrinkle, anti-inflammatory, whitening, and anti-aging skin function improvement effects (Jang Young-A and Lee Jin-Tae, 2015; Chien-Liang Fang et al. , 2020).

Cheonmundong (天門冬) is a perennial herb belonging to the family Liliaceae and has various therapeutic effects such as anti-inflammatory, diuretic, antitussive, antibacterial, nerve stabilization, salivation promotion, and antipyretic. It has been used to treat inflammatory diseases related to hepatitis, asthma, brain disease, and dermatitis with a beneficial effect. As a result of research related to skin physiological activity, skin inflammation inhibitory effect and antioxidant effect are known (Kyungyun Koo et al., 2016).

According to one embodiment of the present invention, the complex extract of frankincense, baechohyang, mulchyang, clove, danhyang, ginseng, licorice, broiler, bokryeong and cheonmundong of the present invention is prepared as follows. First, dried frankincense, baechohyang, mulberry, clove, danhyang, ginseng, licorice, broiler, bokryeong and cheonmundong are mixed to prepare a mixture.

At this time, at least one selected from the group consisting of the bark, gum, and resin of frankincense is used, and at least one selected from the group consisting of outposts, stems, leaves and callus of baechohyang is used, is used, and at least one selected from the group consisting of clove outposts, stems, bark, buds, flowers, buds and leaves is used, and at least one selected from the group consisting of unidirectional outposts, xylem and seeds is used do. In addition, at least one selected from the group consisting of roots, fruits, seeds, leaves, shoots, callus, flowers and stems of ginseng is used, and at least one selected from the group consisting of outposts, roots, shoots, leaves and stems of licorice is used. The bark or leaves of a broiler tree are used, and at least one selected from the group consisting of sclerotia, fruiting body and mycelium of bokryeong is used, and the outpost or root of Cheonmundong is used.

The above frankincense, baechohyang, mulberry, clove, danhyang, ginseng, licorice, broiler chicken, bokryeong and cheonmundong are each 1-2:1-2:1-2:1-2:1-2:1-2:1-2:1-2: It is mixed in a weight ratio of 1 to 2:1 to 2:1 to 2, and it is more preferable because it exhibits the best activity when it is mixed and extracted in the same weight ratio.

Then, at least one extraction solvent selected from the group consisting of water, anhydrous or hydrous alcohol having 1 to 4 carbon atoms, ethyl acetate, acetone, glycerin, ethanol, propylene glycol and butylene glycol is added to the mixture in a conventional method Extracted by pouring, filtered and dried to prepare a complex extract. The complex extract may be prepared by preparing each extract and then mixing in the above ratio.

The frankincense tree, baechohyang, woodhyang, clove, danhyang, ginseng, licorice, broiler, bokryeong, and cheonmundong complex extracts prepared in this way have the synergistic effect of their respective extracts, frankincense, baechohyang, woodhyang, clove and danhyang aromaticity Compared to the plant complex extract and the herbal material complex extract of ginseng, licorice, broiler tree, bokryeong and cheonmundong, it showed superior antibacterial effect, keratinocyte migration promoting effect, melanin production inhibition and collagen production increase effect.

Specifically, the frankincense, baechohyang, mulberry, clove, dandelion, ginseng, licorice, broiler, bokryeong and cheonmundong complex extracts of the present invention do not affect the survival rate of skin-derived cell lines (Test Example 1), Inducing cell membrane damage and death of S. aureus , which can cause lesions (Test Examples 2 and 3), and inhibiting biofilm production of C. acnes (Test Example 4). In addition, the skin keratinocyte migration promoting effect (Test Example 5), the melanin production inhibitory effect (Test Example 6), and the collagen production increasing effect were excellent (Test Example 7). In addition, when these complex extracts were applied to serum and cream formulations, formulation stability was shown at room temperature, refrigeration, and constant temperature conditions (Test Example 8). Therefore, the cosmetic composition containing the complex extract as an active ingredient can be usefully used as a cosmetic for antibacterial, skin regeneration, skin whitening or skin wrinkle improvement.

The complex extract as an active ingredient is contained in an amount of 0.001 to 10% by weight based on the total weight of the cosmetic composition.

The cosmetic composition of the present invention may be prepared in any conventionally prepared formulation, for example, lotion, cream, essence, cleansing foam, cleansing water, feminine cleanser, pack, body lotion, body oil, body gel, shampoo, conditioners, hair conditioners, hair gels, foundations, lipsticks, mascaras, makeup bases, and the like.

[Example]

Hereinafter, to help the understanding of the present invention, examples will be described in detail. However, the following examples are merely illustrative of the contents of the present invention, and the scope of the present invention is not limited to the following examples.

Preparation Examples 1 - 5: Preparation of extracts of frankincense, baechohyang, woodhyang, clove, and dandelion

Purified water as an extraction solvent in each 100 g of dried frankincense resin (Preparation Example 1), pear-chrysanthemum outpost (Preparation Example 2), wood-hyang outpost (Preparation Example 3), clove flower (Preparation Example 4), and unicorn stalk (Preparation Example 5) : Butylene glycol = 70:30 (w/w) 10 times the weight of the mixed solution was added and heated to 80~100℃ in an extractor equipped with a cooling condenser (Cosmos-660, Kyungseo Machinery) to extract 3 times for 2 hours each. .

After extraction by the above method, the mixture was left at room temperature for 3 days, and the precipitate was filtered with 300 mesh filter paper, and the precipitate was filtered twice with Edventec No. 5 filter paper and Whatman GFC 150 mm filter paper to prepare extracts, respectively.

Preparation Examples 6-10: Preparation of ginseng, licorice, broiler tree, bokryeong, cheonmundong extract

Purified water as an extraction solvent in each 100 g of dried ginseng root (Preparation Example 6), licorice root (Preparation Example 7), broiler bark (Preparation Example 8), Bokryeong sclerotia (Preparation Example 9), and Cheonmundong root (Preparation Example 10) : Butylene glycol = 70:30 (w/w) 10 times the weight of the mixed solution was added and heated to 80~100℃ in an extractor equipped with a cooling condenser (Cosmos-660, Kyungseo Machinery) to extract 3 times for 2 hours each. .

After extraction by the above method, the mixture was left at room temperature for 3 days, and the precipitate was filtered with 300 mesh filter paper, and the precipitate was filtered twice with Edventec No. 5 filter paper and Whatman GFC 150 mm filter paper to prepare extracts, respectively.

Examples 1 - 6: Preparation of complex extract

Purified water as an extraction solvent in 100 g of a mixture of dried frankincense resin, baechohyang outpost, woodhyang outpost, clove flower, dandelion outpost, ginseng root, licorice root, broiler bark, bokryeong sclerotia and cheonmundong root in the weight ratio shown in Table 1 below: Butylene glycol = 70:30 (w/w) 10 times the weight of the mixed solution was added, and heated to 80~100℃ in an extractor equipped with a cooling condenser (Cosmos-660, Kyungseo Machinery), and extracted 3 times for 2 hours each.

After extraction by the above method, the mixture was left at room temperature for 3 days, and the precipitate was filtered with 300 mesh filter paper, and the precipitate was filtered twice with Edventec No. 5 filter paper and Whatman GFC 150 mm filter paper to prepare a complex extract.

raw material Example 1 Example 2 Example 3 Example 4 Example 5 Example 6 frankincense One - One One 2 One baecho flavor One - One 2 One One elecampane One - One 2 2 One clove One - One One One 2 unidirectional One - One One One One Ginseng - One One One One 2 licorice - One One 2 One One broiler - One One 2 2 One bokryeong - One One One 2 One Cheonmun-dong - One One One One 2

Test Example 1: Confirmation of cytotoxicity

To determine if the cytotoxicity of keratinocyte cell line HaCaT and fibroblasts in NHDF (normal human dermal fibroblast) the were cultured in DMEM medium supplemented with 10% FBS, respectively in 96 well plate ⁵ × 10 5 and 3 × 10 ⁴ Inoculated at a cell concentration and incubated for 24 hours in a 37° C. 5% CO _{2 incubator.} After incubation, the medium was removed, and the extracts prepared in Preparation Examples and Examples were treated so that the sample concentrations were 1% and 2%, and after culturing for 24 hours, MTT (3-[4,5-dimethylthiazol-2yl]-2, 5-diphenyltetrazoliumboromide, Sigma) solution was added to each well by 100 ml (3 mg/ml), and then incubated for 4 hours. Thereafter, the supernatant was removed, 150 μl of dimethyl sulfoxide was added, and the resulting formazan was dissolved by shaking for 30 minutes, and absorbance was measured at 540 nm using a multimicroplate reader (Molecular device Spectra max190). Cell viability was calculated according to the following formula, and the results are shown in Table 2 below.

Cell viability (%) = absorbance of sample addition group / absorbance of control group × 100

division Cell viability (%) HaCAT NHDF One % 2 % One % 2 % Preparation Example 1 99 97 102 99 Preparation 2 98 96 101 98 Preparation 3 100 98 100 100 Preparation 4 100 100 101 100 Preparation 5 101 99 99 100 Preparation 6 99 97 98 97 Preparation 7 98 100 100 97 Preparation 8 97 99 99 100 Preparation 9 99 97 98 97 Preparation 10 100 99 100 99 Example 1 101 97 102 101 Example 2 100 98 101 100 Example 3 99 97 98 100 Example 4 99 99 101 100 Example 5 102 98 102 99 Example 6 100 101 99 97

As can be seen from the results of Table 2, it was confirmed that there was no effect on cytotoxicity as it showed a cell viability of 95% or more in both Preparation Examples and Examples, and there was no problem in safety.

Test Example 2: S. aureus Confirmation of cell membrane damage induction effect

The S. aureus strain used in the experiment was purchased from the Korea Research Institute of Bioscience and Biotechnology Biological Resources Center (KCTC; Korea Collection for Type Culture), and was used after 2 to 3 consecutive passages as fresh cultures grown in liquid or solid medium. . The extracts prepared in Preparation Examples and Examples were treated to sample concentrations of 1% and 2% in a liquid medium containing the strain, and cultured for 24 hours under aerobic conditions at 35°C. Thereafter, the amount of DNA/RNA leakage due to cell membrane damage of S. aureus was confirmed by filtering with a filter of 0.22 μm pore size and measuring absorbance at 260 nm. The cell membrane damage rate of S. aureus was calculated according to the following formula, and the results are shown in Table 3 below.

Cell membrane damage rate (%) of S. aureus = ((absorbance of sample added group - absorbance of control group) / absorbance of control group) × 100

division S. aureus cell membrane damage (%) One % 2 % Preparation Example 1 13 24 Preparation 2 14 25 Preparation 3 12 22 Preparation 4 15 23 Preparation 5 13 24 Preparation 6 5 13 Preparation 7 8 15 Preparation 8 4 13 Preparation 9 6 17 Preparation 10 7 14 Example 1 28 35 Example 2 19 28 Example 3 36 47 Example 4 30 38 Example 5 32 38 Example 6 31 36 1,2-Hexanediol - 57

As can be seen from the results of Table 3, the composite extract of Examples 1 to 6 of the present invention has excellent efficacy in damaging the cell membrane of the S. aureus strain when compared with each extract of Preparation Examples 1 to 10. there was. The complex extract of the aromatic plant of Example 1 showed better efficacy than the herbal material complex extract of Example 2, and the most excellent cell membrane damage inducing efficacy was confirmed in the complex extract of Example 3 prepared by mixing them all.

Test Example 3: S. aureus Check the antibacterial effect on

After dispensing the S. aureus strain in a 24-well plate, the extracts of Preparation Examples and Examples were treated at concentrations of 1% and 2%, and cultured for 48 hours under aerobic conditions at 35°C. Then, the degree of apoptosis of S. aureus was confirmed by transferring the culture medium to a 96-well plate and measuring the absorbance at 600 nm. The antibacterial effect by promoting the death of S. aureus was calculated according to the following formula, and the results are shown in Table 4 below.

Antibacterial effect on S. aureus (%) = ((absorbance of control group - absorbance of sample addition group) / absorbance of control group) × 100

division Antibacterial effect on S. aureus (%) One % 2 % Preparation Example 1 13 24 Preparation 2 16 29 Preparation 3 15 26 Preparation 4 18 27 Preparation 5 16 26 Preparation 6 4 12 Preparation 7 3 11 Preparation 8 6 13 Preparation 9 8 16 Preparation 10 5 15 Example 1 35 48 Example 2 25 31 Example 3 43 55 Example 4 37 53 Example 5 39 53 Example 6 37 51 1,2-Hexanediol - 61

As can be seen from the results of Table 4, it was confirmed that the composite extract of Examples of the present invention had excellent antibacterial efficacy against S. aureus strains when compared with each extract of Preparation Examples 1 to 10. In particular, in Example 3, which is a complex extract of five aromatic plants and five herbal materials, the best antibacterial effect was exhibited.

Test Example 4: C. acnes Confirmation of the inhibitory effect of biofilm formation

After dispensing the C. acnes strain in a 24-well plate, the extracts of Preparation Examples and Examples were treated at concentrations of 1% and 2%, and cultured for 72 hours under anaerobic conditions at 35°C. Then, the culture medium was removed and the wells were washed with a phosphate buffer solution. After adding 0.1% crystal violet solution to the well and staining for 20 minutes, the well was washed with a phosphate buffer solution, and ethanol was added to dissolve the remaining crystal violet in the well. The degree of biofilm formation of C. acnes was confirmed by transferring the lysate to a 96 well plate and measuring the absorbance at 590 nm. The biofilm formation inhibition rate of C. acnes was calculated according to the following formula, and the results are shown in Table 5 below.

Biofilm formation inhibition rate (%) of C. acnes = ((absorbance of control group - absorbance of sample addition group)/absorbance of control group) × 100

division C. acnes biofilm formation inhibition rate (%) One % 2 % Preparation Example 1 16 26 Preparation 2 18 25 Preparation 3 17 27 Preparation 4 18 26 Preparation 5 17 28 Preparation 6 11 19 Preparation 7 12 18 Preparation 8 12 17 Preparation 9 14 20 Preparation 10 13 18 Example 1 24 35 Example 2 19 30 Example 3 31 45 Example 4 27 38 Example 5 26 38 Example 6 28 39 1,2-Hexanediol - 54

As shown in the results of Table 5, it was confirmed that the composite extracts of Examples 1 to 6 of the present invention had excellent efficacy in inhibiting biofilm formation of C. acnes when compared with the respective extracts of Preparation Examples 1 to 10. . The aromatic plant complex extract of Example 1 showed better efficacy than the herbal material complex extract of Example 2, and it was confirmed that the complex extract of Example 3 prepared by mixing them all showed the best biofilm formation inhibitory effect. .

Test Example 5: Confirmation of keratinocyte migration promotion

In the wound healing process of the skin, keratinocytes and fibroblasts migrate for skin regeneration, and they play an important role in skin restoration by synthesizing various extracellular matrices. In order to check the skin damage recovery and skin regeneration efficacy of the preparations and examples of the present invention, the human keratinocyte line HaCaT was used, and was purchased from ATCC (USA). For cell culture, DMEM containing 100 unit/ml penicillin, 100 μg/ml streptomycin and 10% fetal bovine serum (FBS) was used, and cells were cultured in a humidified incubator maintained _{at 37°C and 5% CO 2} did. HaCaT cell lines were added ^{to a 24-well plate at a rate of 2 × 10 5} cells/well and cultured until the wells were filled. Then, after scraping the bottom of the well with a 200 μl pipette tip to the center line of the well, it was washed with a phosphate buffer solution, and the extracts of Preparation Examples and Examples were treated at concentrations of 1% and 2%. The distance between cells caused by the movement of cells at the site where the cells were scraped was quantified using Image J software, and the results are shown in Table 6 below.

division Intercellular distance (%) One % 2 % Preparation Example 1 89 64 Preparation 2 83 61 Preparation 3 85 62 Preparation 4 88 66 Preparation 5 87 61 Preparation 6 74 54 Preparation 7 78 60 Preparation 8 73 57 Preparation 9 70 52 Preparation 10 74 53 Example 1 62 47 Example 2 57 40 Example 3 49 31 Example 4 54 38 Example 5 55 36 Example 6 53 35

As can be seen from the results of Table 6, it was confirmed that the composite extracts of Examples 1 to 6 of the present invention had excellent efficacy in promoting the migration of keratinocytes when compared to the respective extracts of Preparation Examples 1 to 10. . The herbal material complex extract of Example 2 showed superior efficacy than the aromatic plant complex extract of Example 1, and it was confirmed that the complex extract of Example 3 prepared by mixing them all showed the best keratinocyte migration promoting effect. there was.

Test Example 6: Confirmation of melanin production inhibitory effect

B16F10 cells were inoculated in a 24 well plate at ^{a cell concentration of 4×10 5} , respectively, and cultured at 37° C., 5% CO _{2 in an} incubator for 24 hours. After culture, the medium was removed and washed once with PBS (phosphate buffered saline), and 1 ml of PBS was added to each well. The extract samples of Preparation Examples and Examples 1 to 6 of the present invention were added to the medium at 1% and 2%, respectively, and the cells were treated and then cultured for 72 hours. As a positive control, arbutin was used and the sample was replaced once daily.

In order to confirm the inhibition of melanogenesis, the medium was removed and the cells were washed once with PBS. Then, 500 μl of 1 N NaOH was added to each well, and the melanin generated by shaking for 30 minutes was dissolved, and the absorbance was measured at 405 nm using a multimicroplate reader. Melanin production inhibition rate was calculated according to the following formula and the results are shown in Table 7 below.

Melanogenesis inhibition rate (%) = ((absorbance of control group - absorbance of sample addition group) / absorbance of control group) × 100

division Melanin production inhibition rate (%) One % 2 % Preparation Example 1 21 33 Preparation 2 24 34 Preparation 3 19 32 Preparation 4 20 35 Preparation 5 25 37 Preparation 6 36 48 Preparation 7 35 51 Preparation 8 37 50 Preparation 9 39 52 Preparation 10 37 49 Example 1 51 59 Example 2 62 71 Example 3 67 76 Example 4 64 72 Example 5 63 74 Example 6 63 72 Arbutin 79 -

As shown in the results of Table 7, it was confirmed that the composite extracts of Examples 1 to 6 of the present invention effectively inhibited melanin production when compared with the respective extracts of Preparation Examples 1 to 10. In particular, in Example 3, which is a complex extract of five aromatic plants and five herbal materials, the best melanin production inhibitory effect was exhibited.

Test Example 7: Confirmation of the effect of increasing production of Procollagen type 1

In order to confirm the efficacy of inhibiting the production of procollagen type 1, human fibroblasts, NHDF cells, were inoculated into a 24-well plate and _{cultured at 37° C., 5% CO 2 in an} incubator for 24 hours. Thereafter, the extracts prepared in Preparation Examples and Examples were treated to have sample concentrations of 1% and 2%, and then cultured for 24 hours. Then, the culture medium was collected and the amount of procollagen type 1 was measured using the Procollagen type I c-peptide EIA kit (TAKARA, MK101). On the other hand, the cells adhering to the bottom were washed with PBS and then dissolved with 1N NaOH to measure the total amount of protein, and the amount of procollagen type-1 produced per predetermined protein was calculated. The results are shown in Table 8 below.

division Procollagen type 1 production/total protein One% 2% control 4.8 Preparation Example 1 5.4 5.9 Preparation 2 5.3 6.1 Preparation 3 5.6 6.2 Preparation 4 5.6 6.3 Preparation 5 5.5 6.0 Preparation 6 6.9 7.9 Preparation 7 6.7 7.6 Preparation 8 6.5 7.6 Preparation 9 6.8 8.0 Preparation 10 6.7 7.8 Example 1 7.2 8.1 Example 2 7.8 8.4 Example 3 8.7 9.3 Example 4 8.0 8.7 Example 5 8.2 8.9 Example 6 8.2 8.8

As can be seen from the results of Table 8, it was confirmed that the composite extracts of Examples 1 to 6 of the present invention further increased the production of procollagen type 1 when compared with the respective extracts of Preparation Examples 1 to 10. The herbal material complex extract of Example 2 showed better efficacy than the aromatic plant complex extract of Example 1, and the complex extract of Example 3 prepared by mixing them all showed the best effect of increasing collagen production.

Example 7: Preparation of serum containing complex extract

The serum containing the complex extract of Example 3 was prepared according to a conventional method with the composition and content shown in Table 9 below.

Raw material (wt%) Example 7 Complex extract (Example 3) 1.0 beeswax 1.0 Polysorbate 60 1.5 Sorbitan Sesquiolate 0.5 mineral oil 10.0 Sorbitan Stearate 0.5 lipophilic monostearate glycerin 1.0 stearic acid 1.5 Glyceryl Stearate/PEG-400 Stearate 1.0 propylene glycol 3.0 carboxy polymer 0.1 triethanolamine 0.1 Phenoxyethanol appropriate amount Purified water To 100

Example 8: Preparation of cream containing complex extract

A cream containing the complex extract of Example 3 was prepared according to a conventional method with the composition and content of Table 10 below.

Raw material (wt%) Example 8 Complex extract (Example 3) 1.0 shea butter 2.0 Polysorbate 60 1.5 Sorbitan Sesquiolate 0.8 mineral oil 10.0 dimethicone 3.0 Sorbitan Stearate 0.5 lipophilic monostearate glycerin 1.0 cetearyl alcohol 2.0 Glyceryl Stearate/PEG-400 Stearate 1.0 propylene glycol 3.0 carboxy polymer 0.25 triethanolamine 0.25 Phenoxyethanol appropriate amount Purified water To 100

Test Example 8: Formulation stability confirmation

For the formulations prepared in Examples 7 and 8, stored in an opaque glass container at room temperature (25 ° C), refrigeration (4 ° C), and constant temperature (50 ° C) constant at room temperature, refrigerator and incubator for 12 weeks, and Observation (discoloration, discoloration and separation), stability was confirmed, and the results are shown in Table 11 below.

temperature condition Stability check (12 weeks storage) Example 7 Example 8 discoloration odor detach discoloration odor detach Room temperature (25°C) 0 0 0 0 0 0 Refrigeration (4℃) 0 0 0 0 0 0 constant temperature (50℃) 0 0 0 0 0 0

< Formulation stability grade >

0: No change 1: Minor change 2: Change 3: Extreme change

As shown in Table 11 above, it was confirmed that all formulations of Examples 7 and 8 did not show discoloration, odor and separation under the temperature conditions of 25°C, 4°C, and 50°C, and were stable.

Claims (8)

Frankincense, baechohyang, mokhyang, clove, danhyang, ginseng, licorice, broiler, bokryeong and cheonmundong complex which are extracted and manufactured by mixing frankincense, baechohyang, mokhyang, clove, danhyang, ginseng, licorice, broiler, bokryeong and cheonmundong in the same weight ratio. A cosmetic composition containing the extract as an active ingredient in an amount of 0.001 to 10% by weight based on the total weight of the cosmetic composition. delete delete The cosmetic composition according to claim 1, wherein the cosmetic composition is for antibacterial use. The cosmetic composition according to claim 4, wherein the cosmetic composition has antibacterial activity against Staphylococcus aureus and Cutibacterium acnes. The cosmetic composition according to claim 1, wherein the cosmetic composition is for skin regeneration. The cosmetic composition according to claim 1, wherein the cosmetic composition is for skin whitening. The cosmetic composition according to claim 1, wherein the cosmetic composition is for skin wrinkle improvement.