KR101428651B1 - Cosmetic compositions comprising the extract of Alertris farinosa for anti-oxidation, skin-whitening and anti-inflammation - Google Patents
Cosmetic compositions comprising the extract of Alertris farinosa for anti-oxidation, skin-whitening and anti-inflammation Download PDFInfo
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Abstract
Description
본 발명은 항산화, 미백 및 염증 개선활성이 있는 식물 추출물 및 그 추출물을 유효성분으로 함유하는 화장료 조성물에 관한 것이다.The present invention relates to a cosmetic composition comprising as an active ingredient, a plant extract having antioxidant, whitening and inflammation improving activity and an extract thereof.
트루유니콘(True Unicorn; 한국명:파리노사쥐꼬리풀)은 백합과에 속하는 여러해살이풀로서 플로리다 북부 해안가에서 주로 서식한다. 학명은 Alertris farinosa L.이며 이명으로는 Colicroot, Colicweed, Crow Corn, Unicorn Root, Blazing Star, Mealy Starwort, Starwort Unicorn Root, Unicornplant, Unicorn's Horn, Devil's-Bit, Ague Root, Aloeroot, Huskwort이 있다. 북아메리카 지역의 약용식물 중 하나로 배앓이, 소화불량 및 여성질환의 치료제로 사용되고 뿌리 달임물은 강장제로 사용되며 배앓이, 복부팽만 외에도 히스테리를 완화하는 진정 효과가 있다.
True Unicorn is a perennial plant belonging to the Lily family and lives mainly on the northern coast of Florida. Alertris farinosa L. is the scientific name and Colicot, Colicweed, Crow Corn, Unicorn Root, Blazing Star, Mealy Starwort, Starwort Unicorn Root, Unicornplant, Unicorn's Horn, Devil's-Bit, Ague Root, Aloeroot and Huskwort. It is one of the medicinal plants in North America. It is used as a treatment for colic, indigestion and female diseases. Root moon is used as a tonic. It has a calming effect to alleviate hysteria besides colic and abdominal distension.
자란 및 에델바이스 추출물을 함유하는 화장료 조성물에 관하여는 대한민국 등록특허 제10-0512690호가 개시된 바 있고, 열매지방에서 자생하는 식물 추출물을 함유하는 항노화, 미백, 항염 활성이 있는 화장료 조성물에 관하여는 등록특허 제10-1153320호가 개시된 바 있고, 인플라메이징 개선용 및 피부 노화 방지용 피부 외용제 조성물에 관하여 모과 추출물, 석곡 추출물, 익지인 추출물, 죽엽 추출물, 차전자 추출물, 천문동 추출물, 홍화인 추출물을 함유하는 피부 외용제에 관하여는 등록특허 제10-0539298호가 개시된 바 있다. 그러나 트루 유니콘 추출물을 함유한 화장료 조성물에 관하여는 전혀 개시되거나 암시된 바 없다.Regarding the cosmetic composition containing the extract and the extract of Edelweiss, Korean Patent No. 10-0512690 has been disclosed, and a cosmetic composition having an anti-aging, whitening and anti-inflammatory activity containing a plant extract naturally occurring in a fruit fat, 10-1153320 discloses a composition for external application of skin for improving inflammation and preventing aging of skin. The composition for external application of skin for improving inflammation and skin aging is a skin containing extracts of Quercus, Extract, Ripigrant Extract, Bamboo Extract, Tea Extract, Regarding the external preparation, Patent No. 10-0539298 has been disclosed. However, nothing has been disclosed or implied at all for cosmetic compositions containing truunikon extract.
따라서 본 발명의 목적은 항산화, 미백 및 염증 개선 활성이 있는 트루 유니콘 추출물을 제공하는 데 있다.Accordingly, an object of the present invention is to provide a Truunikon extract having antioxidative, whitening and inflammation-improving activities.
본 발명의 다른 목적은 상기 트루 유니콘 추출물을 유효성분으로 함유하는 항산화, 미백 및 염증 개선용 화장료 조성물을 제공하는 데 있다.Another object of the present invention is to provide a cosmetic composition for improving antioxidant, whitening and inflammation containing the above Unicorn extract as an active ingredient.
본 발명의 상기 목적은 본 발명 트루 유니콘 추출물을 제조하고, 상기 추출물의 항산화활성, 멜라닌 합성 저해활성, Nitric Oxide 저해활성, 비만세포 탈과립 억제활성, 콜라겐 합성능을 확인함으로써 달성하였다. The above object of the present invention is achieved by preparing the Truunikon extract of the present invention and confirming the antioxidative activity, melanin synthesis inhibitory activity, nitric oxide inhibitory activity, mast cell degranulation inhibiting activity and collagen synthesis ability of the extract.
본 발명은 트루 유니콘 추출물의 제조에 초음파 추출법을 사용함으로써 항산화활성 및 멜라닌합성 저해활성이 우수할 뿐만 아니라 항알러지 및 콜라겐 합성에 뛰어난 효과가 있다.The present invention has excellent antioxidative activity and melanin synthesis inhibitory activity by using the ultrasonic extraction method in the production of the Truunikon extract, and is excellent in antiallergic and collagen synthesis.
도 1은 본 발명 트루 유니콘 추출물의 항산화 효과를 나타낸 그래프이다.
도 2는 본 발명 트루 유니콘 추출물의 멜라닌 합성 저해효과를 나타낸 그래프이다.
도 3은 본 발명 트루 유니콘 추출물의 Nitric Oxide 저해효과를 나타낸 그래프이다.
도 4는 본 발명 트루 유니콘 추출물의 비만세포 탈과립 억제효과를 나타낸 그래프이다.
도 5는 본 발명 트루 유니콘 추출물의 콜라겐 합성효과를 나타낸 그래프이다.1 is a graph showing the antioxidative effect of Truunikon extract of the present invention.
FIG. 2 is a graph showing the inhibitory effect on Melanin synthesis of the Truunikon extract of the present invention.
FIG. 3 is a graph showing the inhibitory effect of Nitric Oxide on the Truunikon extract of the present invention.
4 is a graph showing the inhibitory effect of truic unicon extract on mast cell degranulation in the present invention.
5 is a graph showing the collagen synthesis effect of the Truunikon extract of the present invention.
본 발명은 트루 유니콘 추출물을 유효성분으로 함유하는 화장료 조성물을 제공한다.The present invention provides a cosmetic composition comprising Truunikon extract as an active ingredient.
본 발명에 있어서 가장 바람직하기는 상기 트루 유니콘 추출물은 건조된 트루 유니콘에 유기용매를 가하고 sonificator 온도를 40℃로 고정한 후 초음파 처리하여 제조한다. 그러나, 본 발명의 추출물은 물 또는 유기용매 추출만으로도 가능하다. 이때 사용하는 유기용매는 에탄올, 부틸렌글리콜, 프로필렌글리콜, 프로판디올 등이다.In the present invention, most preferably, the Truunikon extract is prepared by adding an organic solvent to the dried Truunicorn, fixing the sonificator temperature to 40 ° C, and subjecting the resultant to ultrasonic treatment. However, the extract of the present invention can be obtained by only water or organic solvent extraction. The organic solvent used herein is ethanol, butylene glycol, propylene glycol, propane diol, and the like.
본 발명에 있어서 상기 제조된 트루 유니콘 추출물은 화장료 조성물 총 중량의 0.1∼10중량％로 바람직하게는 2중량％가 되게 함유하는 것을 특징으로 한다. 0.1중량％ 이하에서는 본 발명의 활성 산소종 저해효과가 미흡하게 나타나며, 10중량％ 이상 사용하는 경우 최종 제품에서 고유 풍미에 따른 고유취와 색상이 강하게 나타나 제품 적용에 어려움이 있다.In the present invention, the Truunikon extract prepared is contained in an amount of 0.1 to 10% by weight, preferably 2% by weight, based on the total weight of the cosmetic composition. When the content is less than 0.1% by weight, the effect of inhibiting the active oxygen species of the present invention is insufficient. When the content is more than 10% by weight, the final product has inherent odor and color depending on its unique flavor.
본 발명에 따른 피부 보호 효과를 가지는 화장료 조성물은 통상의 제형을 가질 수 있으며, 예를 들어 수용성 리퀴드, 로션, 크림, 젤, 팩, 에센스, 수중유(O/W), 유중수(W/O)형 또는 실리콘중수(W/Si)형의 파우더로 제형화될 수 있다. 이들 각 제형의 조성물은 그 제형의 제제화에 필요하고 적절한 각종의 기제와 첨가물을 함유할 수 있으며, 이들 성분의 종류와 양은 당업자에 의해 용이하게 선정될 수 있다.The cosmetic composition having a skin protecting effect according to the present invention may have conventional formulations and may be in the form of, for example, water-soluble liquid, lotion, cream, gel, pack, essence, oil / water, ) Or a silicone heavy (W / Si) type powder. The composition of each of these formulations may contain various kinds of bases and additives necessary for formulation of the formulation, and the kinds and amounts of these ingredients can be easily selected by those skilled in the art.
또한 본 발명에 따른 항산화 효과, 미백 및 염증개선 효과를 가지는 식물 추출물을 함유한 화장료 조성물은 통상적인 사용방법에 따라 사용될 수 있으며, 사용자의 피부상태 또는 취향에 따라 그 사용횟수를 달리할 수 있다.
The cosmetic composition containing the plant extract having the antioxidative effect, whitening effect and inflammation improving effect according to the present invention can be used according to the usual use method and can be used differently depending on the skin condition or taste of the user.
이하, 본 발명의 구체적인 내용을 실시예를 들어 상세히 설명한다.
Hereinafter, the present invention will be described in detail with reference to examples.
실시예Example : : 트루True 유니콘Unicorn 추출물의 제조 Preparation of extract
건조된 트루 유니콘 2∼3g에 40％ 프로판디올(propandiol) 40∼60 mL를 첨가한 후 solificator(JAC 4020, Jinwoo, Korea)를 이용하여 온도를 40℃로 고정한 후 추출하였다. 이렇게 추출된 시료는 0.45μm 막 필터로 여과를 수행하여 본 발명 트루 유니콘 추출물을 얻었다. 모든 과정은 1시간씩 3회 반복하여 실시하였다.
40 ~ 60 mL of 40% propanediol was added to 2 ~ 3g of dried Truunikon and the temperature was fixed at 40 ℃ using a solificator (JAC 4020, Jinwoo, Korea) and then extracted. The thus extracted sample was filtered with a 0.45 μm membrane filter to obtain the Truunikon extract of the present invention. All the procedures were repeated 3 times for 1 hour.
실험예Experimental Example 1: 본 발명 1: invention 트루True 유니콘Unicorn 추출물의 항산화 활성 실험 Antioxidant activity test of extract
본 실험예에서는 피부 섬유아세포(HDFn)를 DMEM(10％ 혈청 첨가, 1％ 항생제 포함) 배지에 현탁하여 37℃, 5％ CO2 배양기에서 배양한 후 대수기에서 성장하는 세포를 실험에 사용하였다. In this experimental example, the skin fibroblast (HDFn) was suspended in DMEM (containing 10% serum and 1% antibiotic), cultured in a 5% CO 2 incubator at 37 ° C, .
Well 당 약 1×105 세포 수가 되도록 96 well에 각각 접종한 후 시료 추출물 0.5％, 1％ 및 2％로 처리한 다음 37℃, 5％ CO2 배양기에서 20분 동안 배양하였다. 산화스트레스 유발물질인 H2O2 1mM을 첨가하고 DCFH-DA (stock 20mM) 20μM를 가한 후 spectrofluorophotometer(excitation 485 nm, emission 535 nm)로 측정하였다. 37℃에서 90분 동안 반응을 관찰하였다. 시료를 넣지 않고 활성 산소종 (Reactive oxygen species)의 형성만 측정한 것을 대조군으로 정하고, 시료를 넣어 활성 산소종을 소거시키는 시료 처치군과 비교하여 활성 산소종 저해율을 구하였다. 모든 실험은 3번 반복하였다.Wells were inoculated in 96 wells at a concentration of about 1 × 10 5 cells per well, treated with 0.5%, 1%, and 2% of sample extracts, and then cultured in a 5% CO 2 incubator at 37 ° C for 20 minutes. 1 mM H 2 O 2 as an oxidative stress inducer was added and 20 μM DCFH-DA (
실험 결과 도 1에 나타난 바와 같이 본 발명 트루 유니콘 추출물의 항산화 활성은 시간에 따라 증가하였다. 상기 트루 유니콘 추출물의 농도가 높을수록 증가율이 높았으며 특히 트루 유니콘 추출물 2％ 처리시 활성 산소종의 형성을 19％ 저해하였다.
Experimental Results As shown in FIG. 1, the antioxidant activity of the truic unicon extract of the present invention increased with time. The higher the concentration of the Truunikon extract, the higher the rate of growth. In particular, the treatment with 2% Truunikon extract inhibited the formation of active oxygen species by 19%.
실험예Experimental Example 2: 본 발명 2: invention 트루True 유니콘Unicorn 추출물의 멜라닌 합성 Melanin synthesis of extract 저해능Low performance 실험 Experiment
본 실험예에서는 murine melanoma(B-16 F1) 세포를 적량의 FBS(fetal bovine serum)가 함유된 DMEM 배지로 6-well plate에 well당 1×105개로 접종한 후 5％ CO2, 37℃하에서 세포가 well 바닥에 약 80％ 이상 부착될 때까지 배양하였다. 배지를 제거하고 시료가 적당 농도로 희석된 배지로 교체한 후 5％ CO2, 37℃하에서 일정시간 배양하였다. The present experiment example, the murine melanoma (B-16 F1) were inoculated the cells a suitable amount of a 6-well plate with DMEM medium containing FBS (fetal bovine serum) 1 × 10 5 pieces per well 5% CO 2, 37 ℃ Were cultured under the condition that the cells were attached to the bottom of the well by about 80% or more. Remove the medium and the sample is incubated under a certain amount of time after the appropriate concentration replaced with a culture medium was diluted with 5% CO 2, 37 ℃.
배지를 제거한 세포를 PBS(phosphated buffer saline)로 세척한 후 트립신으로 처리하여 세포를 회수하고, 회수된 세포는 hematocytometer를 이용하여 세포 수를 측정한 후 5,000∼10,000 rpm으로 10분 동안 원심분리하여 상등액을 제거하고 pellet을 얻었다. Cells were washed with PBS (phosphatized buffer saline), treated with trypsin, and recovered. Cells were recovered by centrifugation at 5,000 to 10,000 rpm at 5,000 to 10,000 rpm using a hematocytometer, And the pellet was obtained.
상기 pellet은 60℃에서 건조한 후 10％ DMSO가 함유된 1M 수산화나트륨액 100μL 또는 적량의 cell lysis buffer를 넣고 60 ℃ 항온조에서 유지한 다음 microplate reader로 405 nm에서 흡광도를 측정하였다.
The pellet was dried at 60 ° C., and then 100 μl of 1 M sodium hydroxide solution containing 10% DMSO or an appropriate amount of cell lysis buffer was added to the pellet. The pellet was maintained in a 60 ° C. incubator and then absorbance was measured at 405 nm with a microplate reader.
실험 결과 도 2에 나타난 바와 같이 본 발명 트루 유니콘 추출물의 농도가 높을수록 멜라닌 합성을 저해하는 활성이 높았으며 특히 상기 추출물 2％ 처리시 대조군에 비해 82％가 저해되었다.
As shown in FIG. 2, the activity of inhibiting melanin synthesis was higher when the concentration of Truunikon extract of the present invention was higher. In particular, the extract of Truunikon was inhibited by 82% when treated with 2% of the extract.
실험예Experimental Example 3: 본 발명 3: invention 트루True 유니콘Unicorn 추출물의 Extract NitricNitric OxideOxide 저해능Low performance 실험 Experiment
본 실험예에서는 Mouse macrophage cell인 RAW 264.7 cell에 염증 유발물질인 LPS를 인위적으로 처리한 후 염증 억제 효과가 있는지를 평가하였다. In this experimental example, RAW 264.7 cell, a mouse macrophage cell, was treated with artificial LPS, which is an inflammation inducing substance, to evaluate whether it has an inflammation inhibitory effect.
96 well plate에 well 당 2×105cell이 되도록 분주한 다음, 24시간 동안 37℃, 5％ CO2 incubator에서 배양하였다. Overnight하고 새로운 배지로 교환한 후 LPS(1ug/mL)와 추출물을 농도별로 투여하여 24시간 동안 배양하였다. 배양 후, 배지 상층액을 취해 13,000 rpm에서 3분 동안 원심 분리하여 상등액만 모아서 Griess 시약과 1:1로 반응시키고 570nm에서 흡광도를 측정하였다.
The cells were plated in 96-well plates at a density of 2 × 10 5 cells per well, and then cultured for 24 hours at 37 ° C. in 5% CO 2 incubator. LPS (1 ug / mL) and extracts were added at different concentrations and incubated for 24 hours. After incubation, the supernatant was taken and centrifuged at 13,000 rpm for 3 minutes. The supernatant was collected, reacted with Griess reagent 1: 1 and absorbance was measured at 570 nm.
실험 결과 도 3에 나타난 바와 같이 본 발명 트루 유니콘 추출물의 농도가 높을수록 Nitric Oxide 저해활성이 높았고 특히 상기 추출물 2％ 처리시 대조군에 비하여 24％가 저해되었다.
Experimental Results As shown in FIG. 3, the higher the concentration of Truunikon extract of the present invention was, the higher the inhibitory activity of Nitric Oxide was. In particular, the treatment of 2% of the extract showed 24% inhibition compared with the control.
실험예Experimental Example 4: 본 발명 4: invention 트루True 유니콘Unicorn 추출물의 비만세포 Mast cell of extract 탈과립Degranulation 억제 실험 Inhibition experiment
본 실험예에서는 RBL-2H3 cell을 10% FBS를 함유한 DMEM(Dulbecco's modified Eagle's medium)에 현탁시킨 후 24 well plate에 각 well당 2×105cell이 들어가도록 처리하였다. 각 well당 100ng/mL의 mouse monoclonal IgE로 감작시킨 후 5％ CO2 incubator에서 24 시간 동안 배양하였다. In this experiment, RBL-2H3 cells were suspended in DMEM (Dulbecco's modified Eagle's medium) containing 10% FBS, and then treated with 2 × 10 5 cells per well in a 24-well plate. Each well was sensitized with 100 ng / mL mouse monoclonal IgE and incubated in a 5% CO 2 incubator for 24 hours.
세포를 well당 Tyrode buffer 500ul로 헹구어내고 농도별로 추출물을 처리한 후 37℃, 5％ CO2 incubator에서 15분 동안 배양하고 1 시간 동안 DNP-BSA 100ng/mL을 처리하였다. 상층액에 substrate (1mM p-nitrophenyl-N-acetyl-ß-D-glucosaminide) 50ul 처리 후 37℃, 5％ CO2 incubator에서 1 시간 동안 incubation 한 후 ELISA reader를 사용하여 405nm에서 흡광도를 측정하였다.
Cells were rinsed with 500 μl of Tyrode buffer and treated with DNP-
실험 결과 도 4에 나타난 바와 같이 본 발명 트루 유니콘 추출물의 농도가 높을수록 비만세포의 탈과립 억제활성이 높았으며 특히 상기 추출물 2％ 처리시 대조군에 비하여 32％가 저해되었다.
As shown in FIG. 4, as the concentration of the Truunikon extract of the present invention was higher, the degranulation inhibitory activity of the mast cells was high, and 32% of the extract of 2% of the extract was inhibited compared to the control.
실험예Experimental Example 5: 본 발명 5: invention 트루True 유니콘Unicorn 추출물의 콜라겐 Collagen of extract 합성능Sum performance 실험 Experiment
본 실험예에서는 피부 섬유아세포(HDFn)를 48 well plate에 5×104cells/well의 농도로 들어가도록 배양 배지 0.3 mL로 접종하여 37℃, 5％ CO2에서 24시간 동안 배양하였다. 그리고 추출물을 농도별로 포함한 새로운 배지에서 24시간 동안 더 배양하였다. 배양액을 모아서 enzymes-linked immunoassay kit (Takara사)를 이용하여 콜라겐 전구체의 C-말단의 양을 측정하고 콜라겐 용액 (640 ng/mL)을 이용하여 콜라겐 농도 0 ng∼640 ng 사이의 standard curve를 얻어 배지에 있는 콜라겐 양을 계산하였다.
In this experiment, HDFn was inoculated into a culture medium (0.3 mL) at a concentration of 5 × 10 4 cells / well in a 48-well plate and cultured at 37 ° C and 5% CO 2 for 24 hours. Then, the extracts were further cultured for 24 hours in a fresh medium containing the concentrations. The cultures were collected and the amount of C-terminus of the collagen precursor was measured using an enzymes-linked immunoassay kit (Takara), and a standard curve between collagen concentration of 0 ng and 640 ng was obtained using collagen solution (640 ng / The amount of collagen in the medium was calculated.
실험 결과 도 5에서 나타난 바와 같이 본 발명 트루 유니콘 추출물의 농도가 증가할수록 콜라겐 합성능도 증가하였고 특히 상기 추출물 2％ 처리시 콜라겐 합성능은 125％로 증가하였다.
As shown in FIG. 5, as the concentration of the Truunikon extract of the present invention was increased, the collagen synthesis performance was increased. In particular, the collagen synthesis performance was increased to 125% when the extract was treated with 2% of the extract.
[산업상 이용가능성] [Industrial applicability]
이상 설명한 바와 같이 본 발명은 트루 유니콘(Alertris farinosa L.) 유래의 추출물을 제공하는 효과가 있을 뿐 아니라 상기 추출물을 유효성분으로 함유하는 항산화, 미백 또는 염증예방 및 개선용 화장료 조성물을 제공하는 뛰어난 효과가 있으므로 화장품 산업상 매우 유용한 발명인 것이다.The present invention as described above is true Unicorn (Alertris farinosa L.), as well as an excellent effect of providing a cosmetic composition for antioxidative, whitening or inflammation prevention and improvement containing the above extract as an active ingredient. Therefore, it is an extremely useful invention in the cosmetics industry.
Claims (4)
Truunikon extract, which is obtained by adding propanediol to Truunikon ( Alertris farinosa L.), ultrasonically extracting it at 40 ° C and filtering it.
The cosmetic composition according to claim 3, wherein the cosmetic composition is a powder selected from the group consisting of a water-soluble liquid, lotion, cream, gel, pack, essence, O / W, W / O or W / Lt; RTI ID = 0.0 > 1, < / RTI >
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