KR101190943B1 - Cosmetic compositions for improving acne containing Oenothera odorata extract - Google Patents

Cosmetic compositions for improving acne containing Oenothera odorata extract Download PDF

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KR101190943B1
KR101190943B1 KR1020040081300A KR20040081300A KR101190943B1 KR 101190943 B1 KR101190943 B1 KR 101190943B1 KR 1020040081300 A KR1020040081300 A KR 1020040081300A KR 20040081300 A KR20040081300 A KR 20040081300A KR 101190943 B1 KR101190943 B1 KR 101190943B1
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evening primrose
acne
reductase
seed extract
primrose seed
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KR20060032372A (en
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김관철
이기무
류근석
김정기
신경훈
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애경산업(주)
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
    • A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
    • A61K8/9783—Angiosperms [Magnoliophyta]
    • A61K8/9789—Magnoliopsida [dicotyledons]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q17/00—Barrier preparations; Preparations brought into direct contact with the skin for affording protection against external influences, e.g. sunlight, X-rays or other harmful rays, corrosive materials, bacteria or insect stings
    • A61Q17/005—Antimicrobial preparations
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q19/00—Preparations for care of the skin
    • A61Q19/008—Preparations for oily skin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
    • A61K2800/74—Biological properties of particular ingredients
    • A61K2800/78—Enzyme modulators, e.g. Enzyme agonists
    • A61K2800/782—Enzyme inhibitors; Enzyme antagonists

Abstract

본 발명은 여드름에 효과가 있는 천연물을 함유하는 화장료 조성물에 관한 것으로, 5α-리덕타아제(5α-Reductase)의 활성을 저해하여 피지의 과잉생산을 억제할 수 있는 달맞이꽃 종자 추출물을 함유하는 여드름의 예방 및 치료용 화장료 조성물에 관한 것이다.
The present invention relates to a cosmetic composition containing a natural product that is effective against acne, and inhibits the activity of 5α-Reductase (5α-Reductase) of the acne containing evening primrose seed extract that can inhibit the overproduction of sebum It relates to a cosmetic composition for prophylaxis and treatment.

5α-리덕타아제, 달맞이꽃, 화장료5α-reductase, evening primrose, cosmetic

Description

달맞이꽃 종자 추출물을 함유하는 여드름 개선 화장료 조성물{Cosmetic compositions for improving acne containing Oenothera odorata extract}Cosmetic composition for improving acne containing Oenothera odorata extract

본 발명은 여드름에 효과가 있는 천연물 함유 화장료 조성물에 관한 것으로, 더욱 상세하게는 여드름 증상을 악화시키는 5알파-리덕타아제(5α-Reductase)의 활성을 강력히 저해하여 피지의 과잉생산을 억제할 수 있는 기능을 가진 달맞이 꽃 추출물을 함유하는 여드름의 치료 및 예방에 효과가 있는 화장료 조성물에 관한 것이다. The present invention relates to a natural-containing cosmetic composition that is effective against acne, and more particularly, to inhibit the excessive production of sebum by strongly inhibiting the activity of 5 alpha-reductase (5α-Reductase) that exacerbates acne symptoms. It relates to a cosmetic composition that is effective in the treatment and prevention of acne containing evening primrose flower extract.

여드름이란 모낭 및 피지선에 발생하는 만성 염증성 질환을 말하는 것으로 이러한 여드름의 발생원인은 크게 피지분비의 증가, 모낭상피의 이상 과각화 및 혐기성 피부 상재균인 프로피오니박테리움 아크네의 증식이 주원인이라 할 수 있으며, 또한 여러 가지 메커니즘이 복합적으로 작용할 수 있다. 남성 호르몬의 작용을 피지 생성 기전과 관련하여 간략히 설명하면 다음과 같다. 남성 호르몬 중 하나인 테스토스테론(testosterone)은 5알파-리덕타아제(5α-Reductase)에 의하여 남성 호르몬의 활성형태인 디하이드로테스토스테론(dihydrotestosterone)으로 전환되고, 이 호르몬의 작용으로 피지가 과잉 분비되어 피지로 인한 질환이 발병된다. 또한, 남성호르몬의 분비상승에 의해 피지선의 기능이 상승되면 생성된 과잉의 피지가 모낭 벽의 과각화로 인하여 모낭 내에 정체되면서 생기는 면포가 여드름의 초기단계이다. 한편, 여드름 환자의 피지 내 지방 성분에도 변화가 일어나 모낭 내 각질형성세포 간의 결합을 증진시켜 모낭정체 과각화증을 초래한다. 그 결과 모낭 내에 정체된 피지는 모낭을 막아 상주하는 혐기성 세균인 프로피오니박테리움 아크네의 생장조건을 형성하여 여드름균(P. acnes)이 지방분해효소와 화학주성인자를 분비하여 유리지방산 및 각종 저분자 물질들을 만들고 백혈구가 모낭주위에 모이게 하여 이들이 모낭 벽을 자극하고 파괴하여 모낭내용물이 진피 내로 유출되어 염증반응이 일어나게 된다. 지금까지 여드름을 치료하기 위해 의약 영역에서 에리드로마이신 등의 항생제를 이용하여 여드름균을 억제하거나 여성호르몬인 에스트로겐을 이용한 피지조절 치료를 수행하였으나 부작용을 가지고 있는 실정이며, 화장료 영역에서는 비타민 A 유도체를 이용한 피부각질 제거 및 벤조일 퍼옥사이드 등을 이용한 여드름균 억제가 사용되어져 왔지만, 이러한 약제 등은 어느 정도의 효과를 나타내는 반면 대부분, 피부발적, 피부과민 반응 또는 광과민 반응 등의 부작용을 일으킬 수 있다는 문제점을 가지고 있다.Acne is a chronic inflammatory disease of hair follicles and sebaceous glands. The causes of acne are mainly due to increased sebum secretion, abnormal hyperkeratosis of the hair follicle epithelium, and proliferation of propionibacterium acne, an anaerobic skin flora. In addition, several mechanisms can work in combination. The action of testosterone is briefly described in relation to the sebum production mechanism as follows. Testosterone, one of the testosterones, is converted to dihydrotestosterone, an active form of testosterone, by 5alpha-Reductase, and the action of this hormone leads to excessive secretion of sebum. Disease due to In addition, when the function of the sebaceous gland is increased by the increase in the secretion of male hormones, the scalp produced by stagnation in the hair follicles due to the hyperkeratosis of the hair follicle wall is an early stage of acne. On the other hand, changes in the fat component of sebum in acne patients promotes binding between keratinocytes in hair follicles, leading to hair follicle hyperkeratosis. As a result, the sebum stagnated in the hair follicles forms the growth conditions of propionibacterium acne, an anaerobic bacterium that clogs the hair follicles, and P. acnes secretes lipolytic enzymes and chemotactic factors to free fatty acids and various low molecular weight molecules. They create substances and cause white blood cells to gather around the hair follicles, causing them to irritate and destroy the hair follicle walls, causing the hair follicle contents to leak into the dermis, causing an inflammatory reaction. Until now, in order to treat acne, in the pharmaceutical area, antibiotics such as erythromycin were used to suppress acne bacteria or sebum control treatment using estrogen, a female hormone, but it has a side effect. Although it has been used to remove acne bacteria and suppress acne bacteria using benzoyl peroxide, such drugs have some effects, but most of them may cause side effects such as skin redness, skin sensitization or photosensitization. Have.

이에 본 발명자는 부작용이 없으며 피부에 자극이 없는 여드름 개선기능을 갖는 물질을 찾기 위하여 국내 자생 식물을 중심으로 검색하였으며, 이 중 달맞이꽃 종자 추출물이 5알파-리덕타아제(5α-Reductase)의 활성을 저해하는 기능이 있음을 발견하여 본 발명을 완성하게 되었다.In this regard, the present inventors searched around domestic native plants to find a substance having an acne-improving function without any side effects, and among them, Evening Primrose seed extracts showed the activity of 5 alpha-Reductase (5α-Reductase). It has been found that there is an inhibitory function to complete the present invention.

본 발명에 따른 여드름 예방 및 치료용 화장료 조성물은 달맞이꽃 종자 추출물을 함유하는 것을 특징으로 한다.Cosmetic composition for preventing and treating acne according to the present invention is characterized in that it contains an evening primrose seed extract.

바람직하게는, 상기 달맞이꽃 종자 추출물을 0.001～10 중량% 함유할 수 있다.Preferably, the evening primrose seed extract may contain 0.001 to 10% by weight.

이하에서 본 발명을 더욱 상세히 설명한다.Hereinafter, the present invention will be described in more detail.

본 발명은 여드름 발생에 있어서 주요한 과정에 있는 피지생성에 관여하는 5α-리덕타아제(5α-Reductase)의 활성을 저해하여 여드름의 예방 및 치료 효과가 있는 달맞이꽃 종자 추출물을 함유하는 화장료 조성물에 관한 것이다. The present invention relates to a cosmetic composition containing an evening primrose seed extract having an effect of preventing and treating acne by inhibiting the activity of 5α-Reductase involved in sebum production in the main process of acne development. .

달맞이꽃(Oenothera odorata)은 바늘꽃과의 두해살이 식물로서 그 추출물은 리놀산과 γ-리놀렌산 및 올레인산 등의 불포화 지방산으로 구성되어있는 것으로 알려져 있으며(Jaocs., Vol.61, March 1984), 한방에서는 월견초(月見草)라는 약재로 쓰이고 있으며, 감기로 열이 높고 인후염이 있을 때 사용하며, 종자는 월견자(月見子)라고 하여 고지혈증 등에 사용해 오고 있는 식물이다. Oenothera odorata is a biennial plant of the family Pinaceae, whose extract is known to consist of unsaturated fatty acids such as linoleic acid, γ-linolenic acid, and oleic acid (Jaocs., Vol. 61, March 1984).月見草) is used as a medicine, high fever due to colds and sore throat is used, and the seeds are moon dogs (月 見 子) is a plant that has been used for hyperlipidemia.

달맞이꽃 종자 추출물은 달맞이꽃 종자를 분쇄하여 80% 에탄올에 3일간 냉침하여 얻었으며, 얻어진 용액을 감압 증발시켜 용매를 제거한 뒤에 동결 건조기에서 건조하여 가루 상태로 얻었다.Evening primrose seed extract was obtained by crushing the evening primrose seeds by cooling in 80% ethanol for 3 days, the resulting solution was evaporated under reduced pressure to remove the solvent and dried in a freeze dryer to obtain a powder.

본 발명의 화장료 조성물은 달맞이꽃 종자 추출물을 0.001 내지 10 중량% 함유하는 것이 바람직하다. 달맞이꽃 종자 추출물의 함량이 0.001중량% 미만일 경우에는 여드름의 예방 및 치료 효과가 미비하고, 10중량%을 초과하는 경우에는 제형 화의 어려움과 피부 부작용의 문제가 있어 적합하지 않다.The cosmetic composition of the present invention preferably contains 0.001 to 10% by weight of evening primrose seed extract. Evening primrose seed content of less than 0.001% by weight is not suitable for the prevention and treatment of acne, when the content exceeds 10% by weight is not suitable because of problems of formulation and skin problems.

한편, 본 발명에 따른 상기 추출물은 유연수, 크림, 로션 등의 화장료 조성물로 사용할 수 있으며, 이들의 제조방법은 공지방법을 따른다.On the other hand, the extract according to the present invention can be used as a cosmetic composition such as soft water, cream, lotion, the production method of these follows a known method.

상기 추출물로 표시되는 유효성분의 사용량은 여드름의 형태, 크기, 부위, 연령에 따라 다양화될 수 있지만, 일반적으로 1일 2회 또는 수회 범위 내에서 사용한다.The amount of the active ingredient represented by the extract may vary depending on the form, size, site, age of acne, but is generally used twice or several times a day.

실험예 1 : 달맞이꽃 종자 추출물의 제조Experimental Example 1 Preparation of Evening Primrose Seed Extract

달맞이꽃 종자 추출물은 달맞이꽃 종자를 분쇄하여, 80% 에탄올을 종자 양의 2배 정도의 부피만큼 가하여 2시간 동안 초음파로 처리한 후에 용액을 따라내는 과정을 2번 반복하였다. 다시 80% 에탄올을 종자에 가하여 3일간 냉침시킨 뒤에 용액을 따라내었다. 상기 과정들에서 얻어진 용액을 모두 합쳐서 증발기를 이용하여 용매를 제거한 뒤에 동결 건조기에서 건조하여 48시간 이상 건조시켜 분말 상태로 만들었다.Evening primrose seed extract was crushed the evening primrose seed, soaking 80% ethanol in a volume of about 2 times the amount of seed and sonicated for 2 hours, followed by the process of decanting the solution twice. Again 80% ethanol was added to the seeds and allowed to cool for 3 days before the solution was decanted. All the solutions obtained in the above processes were combined to remove the solvent using an evaporator and then dried in a freeze dryer to be dried for at least 48 hours to obtain a powder state.

실험예 2 : 5α-리덕타아제 억제력 평가Experimental Example 2 Evaluation of 5α-Reductase Inhibition

달맞이꽃의 5α-리덕타아제 활성 억제력을 확인하기 위해 사용된 5α-리덕타아제를 얻는 과정은 0～2℃에서 수행하였다. 본 시험에 사용된 쥐는 스파라그-돌리(Sprague-Dawley)계로서 290～312g의 무게를 갖는 성숙한 암컷(7-8주령)을 디에틸 에테르로 호흡 마취하고 경추탈골하여 죽인 후, 해부하여 간을 적출하고 완충액(0.13M 수크로오스, 10mM Tris-HCl, 40ul/L 2-머캅토에탄올)으로 세척 후 완충액에 넣어 분쇄기로 현탁액을 조제하였다. 상기 현탁액에서 5α-리덕타아제를 분리하기 위하여 원심분리(105,000g, 1hr,4℃)한 후 상층액에서 취하여 -70℃에서 보관하여 사용하였다.The process of obtaining 5α-reductase used to confirm the inhibition of 5α-reductase activity of evening primrose was carried out at 0 ~ 2 ℃. The rats used in this study were Sprague-Dawley strains and mature females (7-8 weeks old) weighing 290-312g were respiratoryly anesthetized with diethyl ether, killed by cervical vertebral bone, and dissected. The mixture was extracted, washed with buffer (0.13M sucrose, 10 mM Tris-HCl, 40ul / L 2-mercaptoethanol), and then put in a buffer to prepare a suspension by a grinder. To separate 5α-reductase from the suspension, centrifugation (105,000g, 1hr, 4 ° C) was taken from the supernatant and stored at -70 ° C.

5α-리덕타아제 활성억제 효과를 평가하기 위해 SUN ZU-YUE et al.(Indian Journal of Pharmacology, 1998)가 사용한 방법을 사용하였다. 상기와 같은 방법으로 얻은 상층액의 일부를 취하여 완충액 내에서 NADPH2 (조효소), 테스토스테론(기질), 추출물(저해제; inhibitor)과 반응시켜 조효소로 작용하는 NADPH2의 감소량의 변화를 UV 분광광도계(spectrophotometer)를 이용하여 340nm에서 측정하였다. 이 데이터를 이용하여 반응속도, 테스토스테론(기질), 추출물(저해제)이 관련된 Lineweaver-Burk Plot, Dixon Plot을 작성하였으며, 이를 토대로 Ki (저해상수)를 계산하였다. Ki는 반응속도 Vmax와 반포화상수 Km을 1/2로 만드는 저해제의 농도로서, 그 값이 작으면 작을수록 저해 효과가 뛰어남을 의미한다. 표 1은 달맞이꽃 종자추출물의 Ki 값과 대조군의 값을 나타낸다.To assess the effect of 5α-reductase activity inhibition, SUN ZU-YUE et al. The method used by (Indian Journal of Pharmacology, 1998) was used. A portion of the supernatant obtained by the above method was taken and reacted with NADPH 2 (coenzyme), testosterone (substrate), and extract (inhibitor; inhibitor) in the buffer to change the amount of reduction of NADPH 2 that acts as a coenzyme. It was measured at 340nm using a spectrophotometer). Using this data, we prepared Lineweaver-Burk Plot and Dixon Plot related to reaction rate, testosterone (substrate), and extract (inhibitor). Based on this, Ki (low resolution constant) was calculated. Ki is the concentration of the inhibitor that makes the reaction rate V max and the half saturation constant K m 1/2, and the smaller the value, the better the inhibitory effect. Table 1 shows Ki value of evening primrose seed extract and the value of the control group.

구분division Ki (% w/v)Ki (% w / v) 아젤라익산(azelaic acid)Azelaic acid 0.00870.0087 달맞이꽃 종자 추출물Evening Primrose Seed Extract 0.0003100.000310

실험 결과, 양성 대조군(Positive control)으로 사용된 아젤라익산(azelaic acid)보다 뛰어난 5α-리덕타아제 저해 효과가 있음을 확인할 수 이었다.As a result, it was confirmed that the 5α-reductase inhibitory effect is superior to the azelaic acid (azelaic acid) used as a positive control (Positive control).

실험예 3 : 인체 피지선 세포 5α - 리덕타아제에 대한 각 추출물의 활성 저해실험Experimental Example 3: Inhibition of activity of each extract against human sebaceous gland cells 5α-reductase

5α-리덕타아제의 종별, 아종별 저해 효과 차이를 알아보기 위해 일차 배양된 인체 피지선 세포에서 위와 같은 방법으로 5α-리덕타아제를 추출하였다. 피부 조직에는 두 가지 형태(타입 I, 타입 II)의 5α-리덕타아제가 존재하며 이들 두 단백질은 서로 약 60% 유사성(homology)을 갖고 있다. 사람의 경우, 정상 피부에는 타입 I가 타입 II에 비하여 더욱 현저히 발현되지만 타입 II는 주로 전립선에 특이적으로 분포하고, 흰쥐 간의 5α-리덕타아제의 분포 양상은 인체 비생식선 피부(nongenital skin)내의 것과 유사하다. 5α-reductase was extracted from primary cultured human sebaceous gland cells in the same manner as above to determine the different inhibitory effects of 5α-reductase. There are two forms (type I, type II) of 5α-reductase in skin tissue, and these two proteins have about 60% homology with each other. In humans, type I is more pronounced in normal skin than type II, but type II is mainly distributed in the prostate, and the distribution of 5α-reductase in rats is characterized by the presence of nongenital skin in humans. Similar to

여드름과 관련하여 피지 생성에 주요한 역할을 하는 타입 I에 대한 활성 저해효과를 알아보기 위해 인체 피지선 세포를 일차 배양하였으며 주지에(Jujie) 등 (Arch. Dermatol. Res., 1996; 288, 703-708)의 방법을 이용하였다. 피지선 세포(sebaceous gland cell)는 진피층 내의 모낭(hair follicle)에서 얻었으며(영동세브란스 병원 협조), 분리된 피지선 세포는 0.25% 트립신(trypsin), 0.05% 에틸렌디아민테트라아세트산(EDTA)으로 30분간 37℃에서 처리하고, 12% 우태혈청(Fetal Bovine Serum: FBS)이 함유된 EMEM(Eagle's minimum essential medium)으로 트립신을 불활성화시킨 후, 원심 분리를 통해 배지를 제거하였으며, 그 후 KGM(keratinocyte growth medium) 배지에 이분산시켜 60mm 배양용 접시(dish)에서 배양하였다. 1～2회 계대배양을 통하여 100mm 배양용 접시에 배양한 후, 70～80% 정도 자랐을 때 5-알파-리덕타아제를 분리하기 위하여 배양을 종료하였다.(약 2X106 cells/dish당). 배양된 세포 약 1X107 cells (100mm dish 6개)을 수집하여 완충액 (10mM potassium phosphate, 150mM KCl, 1mM EDTA)을 첨가하여 분산시킨 후, 분쇄기로 현탁화하였다. 현탁액은 초음파 파쇄기(sonicator)로 총 3회, 각각 1분씩 처리하여 파괴하고, 원심분리하여 (20,000G) 상층액을 취해 5α-리덕타아제를 부분 정제하여 이 효소의 활성 억제 실험을 수행하였다. 효소의 활성 저해 측정 방법은 실험예 2와 같으며, 실험 결과를 표 2에 나타내었다. Human sebaceous gland cells were first cultured to investigate the inhibitory effect on type I, which plays a major role in sebum production in relation to acne.Jujie et al. (Arch. Dermatol. Res., 1996; 288, 703-708 ) Method was used. Sebaceous gland cells were obtained from hair follicles in the dermal layer (cooperative with Youngdong Severance Hospital), and isolated sebaceous gland cells were treated with 0.25% trypsin and 0.05% ethylenediaminetetraacetic acid (EDTA) for 30 minutes. After treatment at ℃, trypsin was inactivated with EMEM (Eagle's minimum essential medium) containing 12% Fetal Bovine Serum (FBS), the medium was removed by centrifugation, and then KGM (keratinocyte growth medium) ) Was cultured in a 60 mm culture dish. After culturing in a 100 mm petri dish through one or two passages, the culture was terminated to separate 5-alpha-reductase when grown at 70-80% (about 2 × 10 6 cells / dish). About 1 × 10 7 cells (6 100 mm dish) were cultured and dispersed by adding buffer (10 mM potassium phosphate, 150 mM KCl, 1 mM EDTA), and suspended in a grinder. The suspension was destroyed by treating with a sonicator three times in total, 1 minute each, centrifuged (20,000G), and the supernatant was taken to partially purify 5α-reductase to carry out the activity inhibition experiment of this enzyme. The enzyme activity inhibition measurement method is the same as in Experimental Example 2, the experimental results are shown in Table 2.

구분division Ki(% w/v)Ki (% w / v) 아젤라익산(azelaic acid)Azelaic acid 0.00180.0018 달맞이꽃 종자 추출물Evening Primrose Seed Extract 0.00110.0011

실험 결과, 양성대조군으로 사용한 아라젤릭산의 효소 활성 저해효과와 대등한 수준의 효과를 나타내었으며, 이는 인체의 타입 Ⅰ 5α-리덕타아제의 활성 억제제로 달맞이꽃 종자 추출물이 사용 가능함을 보여준다.The experimental results showed that the effect of the enzyme activity of azelalic acid used as a positive control was comparable to that of the enzyme, and that Evening Primrose seed extract can be used as an inhibitor of the type I 5α-reductase activity of the human body.

실험예 4 : 인체 피지선 세포에서의 피지 생성 억제 평가 실험Experimental Example 4: Evaluation test for sebum production in human sebaceous gland cells

인체 타입 I 5-알파-리덕타아제의 활성 억제 효과가 확인된 달맞이꽃 종자 추출물 시험관내(in vitro) 상에서 직접적으로 피지 생성을 어느 정도 억제하는지 확인하고자 피지 생성(lipogenesis) 실험을 수행하였다. 인체 피지선 세포의 수득 및 일차 배양 방법은 실험예 3과 같으며, 계대배양시 본 실험을 위해 24 웰-플레이트(24 well-plate)를 이용하여 웰 당 2X105 개 세포를 심었다. 24시간 배양하여 세포가 웰에 고착된 후, 웰에 달맞이꽃 종자 추출물을 디메틸술폭사이드(dimethylsulfoxide: DMSO)에 분산시켜 최종 농도 0.05%(w/v)로 처리하여 72시간을 배양한 후, 생존한 세포의 수를 계산하고, 생성된 피지의 양을 메탄올:클로로포름 추출법을 이용하여 추출 및 측정하여 104 cell당 생성된 총 피지량을 비교하여 피지생성 억제 정도를 상대 비교하였다. 피지의 추출 방법은 조보울리스(Zoboulis) 등(J. Invest. Dermatol., 1991; 24, 69-83)의 방법을 이용하였으며, 메탄올:클로로포름 추출단계, 초음파 파쇄단계, 필터링 단계, 질소 증발(nitrogen evaporation) 단계를 포함한다. 본 실험 결과를 표 3에 나타내었다.Evening evening primrose seed extract confirmed that the inhibitory effect of the human body type I 5-alpha-reductase activity in vitro (lipogenesis) was performed to determine the degree of sebum production. The method of obtaining and primary culture of human sebaceous gland cells is the same as Experimental Example 3, 2X10 5 cells per well were planted using 24 well-plates for this experiment during subculture. After 24 hours of incubation, the cells were fixed in the wells, and the Evening Primrose seed extract was dispersed in dimethylsulfoxide (DMSO), treated at a final concentration of 0.05% (w / v), incubated for 72 hours, and then The number of cells was counted, and the amount of sebum produced was extracted and measured using methanol: chloroform extraction to compare the total amount of sebum produced per 10 4 cells. Sebum extraction was performed by Zoboulis et al. (J. Invest. Dermatol., 1991; 24, 69-83), methanol: chloroform extraction step, ultrasonic crushing step, filtering step, nitrogen evaporation ( nitrogen evaporation). The results of this experiment are shown in Table 3.

최종 농도 0.05% w/v 처리시At a final concentration of 0.05% w / v 구분division 피지생성 저해율 (%)Sebum production inhibition rate (%) 아젤라익산(azelaic acid)Azelaic acid 3131 달맞이꽃 종자 추출물Evening Primrose Seed Extract 3737

실험 결과, 달맞이꽃 종자 추출물은 인체 피지선 세포의 피지생성을 저해하는 효과가 양성대조군에 비하여 높은 것으로 확인되었으며, 피지생성억제 목적으로 사용 가능한 원료로 판단되어진다.As a result of the experiment, Evening Primrose seed extract was found to have a higher effect of inhibiting sebum production of human sebaceous gland cells compared to the positive control group, and is considered a raw material that can be used for the purpose of inhibiting sebum production.

실험예 5 : 피지분비, 여드름 개선 임상 시험Experimental Example 5: Sebum secretion, acne improvement clinical trial

달맞이꽃 종자유를 함유하는 스킨의 피지분비 억제효과와 여드름 개선효과를 알아보기 위하여 임상시험을 시행하였다. 대상자는 얼굴에 여드름이 난 16～19세의 여성으로 하였으며, 실험군 및 대조군 각각 20명으로 하여 1일 2회 아침세안 후와 저녁 세안 후, 4주 동안 사용케 하였다. 실험군은 실시예 1의 스킨을 사용하였으며 달맞이 꽃 종자유 추출물을 3% 첨가하였다. 대조군은 비교예 1의 스킨을 사용하였다. 실험군 및 대조군의 피지 분비량을 피지량측정기(Sebumeter)를 이용하여 측정하여, 그 결과를 표 4에 나타내었고, 여드름 개선효과를 설문과 육안, 사진촬영으 로 관찰하였으며, 그 결과를 표 5에 나타내었다.A clinical trial was conducted to investigate the sebum secretion and acne-improving effects of skin containing evening primrose oil. The subjects were 16-19 year old females with acne on their faces. Twenty patients in each of the experimental and control groups were used twice a day after morning and evening face washes for 4 weeks. The experimental group used the skin of Example 1 and added 3% of Evening Primrose oil seed extract. As a control, the skin of Comparative Example 1 was used. Sebum secretion of the experimental group and the control group was measured using a sebometer, and the results are shown in Table 4, and the acne improvement effect was observed by questionnaire, visual observation, and photographing, and the results are shown in Table 5. .

피지분비 감소효과Sebum secretion reduction effect 구분division 피지 분비량(사용전) ug/cm3 Sebum secretion (before use) ug / cm 3 피지 분비량(사용후) ug/cm3 Sebum secretion (after use) ug / cm 3 피지분비감소율Sebum secretion rate 시험군Test group 210210 165165 21.4 %21.4% 대조군Control group 213213 196196 8.0 %8.0%

여드름 개선효과Acne Improvement 방법Way 구분division 효과 우수Excellent effect 효과 중간Effect medium 효과 없음no effect 악화worse 부작용Side Effect 설문survey 시험군Test group 1111 88 1One 00 00 대조군Control group 00 44 1414 1One 1One 육안 관찰및 사진 촬영Visual observation and photography 시험군Test group 1212 77 1One 00 00 대조군Control group 00 44 1515 00 1One

표 4와 표 5의 결과를 종합하여 볼 때, 달맞이 꽃 종자유 추출물을 함유한 스킨이 피지분비를 감소시키고, 여드름개선효과가 뛰어난 것으로 나타났다.  In combination with the results of Table 4 and Table 5, the skin containing the Evening Primrose oil extract was found to reduce sebum secretion, excellent acne improvement effect.

실험예 5 : 피부에 대한 안전성 시험Experimental Example 5: Safety Test for Skin

피부에 대한 자극 정도를 평가하기 위하여 첩포 시험을 시행하였다. 피부 첩포시험은 20세 이상의 정상인 30명을 대상으로 실시하였으며, 각각의 시료를 3% 수용액으로 만든 다음 헤이 챔버(Haye's chamber)를 이용하여 48시간 동안 부착한 뒤 제거한 후에 첫 판독을 하고, 96시간이 경과한 후에 2차 판독을 시행하여 피부 반응 결과를 판정하였다. 평가 기준과 자극지수의 계산은 다음과 같으며 그 결과를 표 6에 나타내었다.Patch tests were performed to evaluate the degree of irritation to the skin. The skin patch test was conducted on 30 people over 20 years of age. Each sample was made into a 3% aqueous solution, attached for 48 hours using a Haye's chamber, and then removed for the first reading. After this, a second reading was performed to determine the skin reaction results. Evaluation criteria and stimulation index calculations are as follows and the results are shown in Table 6.

반응reaction 가중치weight 평가 기준Evaluation standard -- 0.00.0 무반응No response ±± 0.50.5 의심되는 양성 반응Suspected positive reaction ++ 1.01.0 약양성반응Weak positive reaction ++++ 2.02.0 강양성반응Strong positive reaction ++++++ 3.03.0 극양성반응Polar positive reaction

피부자극지수=∑(가중치 X 반응인 수)/(최고 가중치 X 총 피험자 수) X 100 Skin irritation index = ∑ (weighted number of responders) / (highest weight X total number of subjects) X 100

시료 ( 3% 수용액 )Sample (3% aqueous solution) 피부자극 지수Skin irritation index 자극의 구분Classification of stimuli 증류수Distilled water 00 무자극No stimulation 달맞이꽃 종자 추출물Evening Primrose Seed Extract 0.280.28 무자극No stimulation

실험결과, 달맞이꽃 종자 추출물은 피부에 대하여 무자극인 것으로 나타났으며, 화장료의 원료로 사용가능하고 피부에 자극이 없으며 여드름 치료 및 예방 기능을 가진 원료로 판단되어진다.As a result of the experiment, evening primrose seed extract was found to be non-irritating to the skin. It can be used as a raw material for cosmetics, has no irritation to the skin, and is considered a raw material with acne treatment and prevention functions.

실시예 1 내지 3, 및 비교예 1Examples 1 to 3, and Comparative Example 1

화장료 조성물Cosmetic composition

상기 실험 결과에 따라 달맞이꽃 종자 추출물을 일정 중량 함유하는 화장수, 크림, 로션제형 화장료를 하기 표 5 내지 7과 같은 조성으로 하여 제조하였다. 각 제형의 화장료 조성물은 반드시 아래 표의 실시예 조성에 국한되는 것은 아니다.According to the experimental results, a lotion, cream, lotion-containing cosmetics containing a certain weight of evening primrose seed extract was prepared by the composition shown in Tables 5 to 7. The cosmetic composition of each formulation is not necessarily limited to the example compositions in the table below.

화장수toilet water 성분ingredient 실시예1(함량%)Example 1 (content%) 비교예1(함량%)Comparative Example 1 (content%) 살리실산Salicylic acid 0.50.5 0.50.5 1,3-부틸렌 글리콜1,3-butylene glycol 5.05.0 5.05.0 구연산Citric acid 0.10.1 0.10.1 폴리옥시에틸렌올레일에테르Polyoxyethylene oleyl ether 0.80.8 0.80.8 에탄올ethanol 30.030.0 30.030.0 향료Spices 0.150.15 0.150.15 방부제antiseptic 적량Suitable amount 적량Suitable amount 달맞이꽃 종자유Evening Primrose Oil 0.001 ~ 100.001 to 10 -- 정제수Purified water 잔량Balance 잔량Balance

크림cream 성분ingredient 함량(중량 %)Content (% by weight) 마카데미아 오일Macadamia oil 2.02.0 스쿠알란Squalane 3.03.0 스테아린산Stearic acid 7.07.0 바셀린vaseline 3.03.0 1,3-부틸렌글리콜1,3-butylene glycol 7.07.0 모노스테아린산 폴리옥시 에틸렌소르비탄Monostearic acid polyoxy ethylene sorbitan 1.31.3 자기유화형 모노스테아린산 글리세린Self-emulsifying glycerin monostearate 2.72.7 에틸렌디아민테트라아세트산 나트륨Ethylenediaminetetraacetic acid sodium 0.050.05 글리세린glycerin 4.04.0 향료Spices 0.250.25 방부제antiseptic 적량Suitable amount 산화방지제Antioxidant 적량Suitable amount 달맞이꽃 종자 추출물Evening Primrose Seed Extract 0.001 ~ 100.001 to 10 정제수Purified water 잔량Balance 합계Sum 100100

로션Lotion 성분ingredient 함량( 중량 % )Content (weight%) 스테아린산Stearic acid 0.50.5 에탄올ethanol 10.010.0 바셀린vaseline 1.01.0 스쿠알란Squalane 2.52.5 유동파라핀Liquid paraffin 6.06.0 모노올레인산폴리옥시에틸렌소르비탄Monooleic acid polyoxyethylene sorbitan 1.51.5 1,3-부틸렌글리콜1,3-butylene glycol 8.08.0 에틸렌디아민테트라아세트산나트륨Sodium Ethylenediaminetetraacetic Acid 0.050.05 트리에탄올아민Triethanolamine 0.80.8 향료Spices 0.20.2 방부제antiseptic 적량Suitable amount 산화방지제Antioxidant 적량Suitable amount 달맞이꽃 종자 추출물Evening Primrose Seed Extract 0.001 ~ 100.001 to 10 정제수Purified water 잔량Balance 합계Sum 100100

상기에서 상술한 바와 같이, 본 발명에 따른 달맞이꽃종자추출물 추출물은 5α-리덕타아제의 활성억제 효과가 뛰어나 피지분비를 억제하며 여드름 예방 및 개선에 매우 유용하다.As described above, the Evening Primrose seed extract according to the present invention is excellent in the inhibitory effect of 5α-reductase, inhibits sebum secretion and is very useful for preventing and improving acne.

Claims (3)

달맞이꽃 종자 추출물을 함유하며, 상기 달맞이꽃 종자 추출물은 5알파 리덕타아제 활성 저해 기능을 갖는 것을 특징으로 하는 여드름 예방 및 개선용 화장료 조성물.Evening primrose seed extract, the evening primrose seed extract is a cosmetic composition for preventing and improving acne, characterized in that it has a 5-alpha reductase activity inhibiting function. 제 1 항에 있어서, 상기 달맞이꽃 종자 추출물을 0.001～10 중량% 함유함을 특징으로 하는 여드름 예방 및 개선용 화장료 조성물.The cosmetic composition for preventing and improving acne according to claim 1, wherein the evening primrose seed extract contains 0.001 to 10% by weight. 제 1 항에 있어서, 상기 달맞이꽃 종자 추출물은 달맞이꽃 종자를 분쇄하고, 에탄올로 추출하여 용매를 증발시킨 후 동결건조시킨 것임을 특징으로 하는 여드름 예방 및 개선용 화장료 조성물.The method of claim 1, wherein the evening primrose seed extract is crushed by evening primrose seed, extracted with ethanol evaporated the solvent and then freeze-dried, characterized in that the cosmetic composition for improvement.
KR1020040081300A 2004-10-12 2004-10-12 Cosmetic compositions for improving acne containing Oenothera odorata extract KR101190943B1 (en)

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