KR102418703B1 - Composition for Improving for Preventing or Treating in Skin Wrinkle and Atopic Dermatitis Comprising Extract of Thymus vulgaris Fermented by Lactic Acid Bacteria as Active Ingredient - Google Patents
Composition for Improving for Preventing or Treating in Skin Wrinkle and Atopic Dermatitis Comprising Extract of Thymus vulgaris Fermented by Lactic Acid Bacteria as Active Ingredient Download PDFInfo
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- KR102418703B1 KR102418703B1 KR1020200009199A KR20200009199A KR102418703B1 KR 102418703 B1 KR102418703 B1 KR 102418703B1 KR 1020200009199 A KR1020200009199 A KR 1020200009199A KR 20200009199 A KR20200009199 A KR 20200009199A KR 102418703 B1 KR102418703 B1 KR 102418703B1
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- lactic acid
- atopic dermatitis
- production
- acid bacteria
- extract
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Abstract
본 발명은 유산균 발효 타임 추출물을 유효성분으로 함유하는 아토피 피부염 및 피부주름 개선용 조성물에 관한 것이다.
본 발명의 유산균 발효 타임 추출물은 아토피 인자인 TARC 및 MDC 단백질의 생성을 효과적으로 억제하고, IL-6와 같은 염증성 사이토카인 생성을 억제하여 아토피성 면역 또는 염증 반응의 억제에 매우 효과적이며, MMP-1 단백질 생성 억제 및 타입 1 프로콜라겐 단백질 생성 증진을 통해 피부 주름의 예방 및 개선에도 효과적이므로, 아토피 피부염 및 피부주름 개선을 위한 화장료, 식품, 및 약학적 조성물로 유용하게 이용될 수 있다.
또한, 유산균 발효 타임 추출물은 천연물인 타임 추출물을 유산균으로 발효한 발효추출물로 인체에 매우 안전할 뿐만 아니라, 안정성도 매우 탁월하다. The present invention relates to a composition for improving atopic dermatitis and skin wrinkles, comprising a lactic acid bacteria fermented thyme extract as an active ingredient.
The lactic acid bacteria fermented thyme extract of the present invention effectively inhibits the production of TARC and MDC proteins, which are atopic factors, and is very effective in inhibiting atopic immunity or inflammatory response by inhibiting the production of inflammatory cytokines such as IL-6, MMP-1 Since it is also effective in preventing and improving skin wrinkles by inhibiting protein production and enhancing the production of type 1 procollagen protein, it can be usefully used as a cosmetic, food, and pharmaceutical composition for improving atopic dermatitis and skin wrinkles.
In addition, lactic acid bacteria fermented thyme extract is a fermented extract obtained by fermenting thyme extract, a natural product, with lactic acid bacteria, which is not only very safe for the human body, but also has excellent stability.
Description
본 발명은 타임 추출물을 유효성분으로 하는 아토피 피부염 및 피부주름 개선용 조성물에 관한 것으로, 더욱 구체적으로 유산균 발효 타임 추출물을 유효성분으로 함유하는 아토피 피부염 및 피부주름 개선용 조성물에 관한 것이다.The present invention relates to a composition for improving atopic dermatitis and skin wrinkles comprising a thyme extract as an active ingredient, and more particularly, to a composition for improving atopic dermatitis and skin wrinkles containing a lactic acid bacterium fermented thyme extract as an active ingredient.
아토피성 피부염(atopic dermatitis)은 만성적으로 재발하는 가려움성 피부염으로, 영유아기에 흔히 발생하고 환자나 가족 중에 가려움증, 피부건조증 또는 특징적인 습진을 동반하는 질환이다. 아토피성 피부염의 전형적인 증상은 손, 두피, 얼굴, 목, 팔꿈치, 무릎 등에서 나타나며, 피부가 매우 건조해지고 가려움증과 염증이 생기며 비늘처럼 벗겨지다가 심하게 긁으면 피부가 두꺼워지면서 깊게 주름이 잡히는 태선화 현상이다.Atopic dermatitis (atopic dermatitis) is a chronic, recurrent itchy dermatitis, a disease that occurs frequently in infancy and is accompanied by itching, dry skin, or characteristic eczema in patients or their families. Typical symptoms of atopic dermatitis appear on the hands, scalp, face, neck, elbows, and knees, and the skin becomes very dry, itchy and inflamed, peels off like scales, and when severely scratched, the skin thickens and wrinkles are formed.
아토피 피부염의 원인은 아직까지 명확히 규명되어 있지 않으나, IgE의 비정상적 증가, 세포 면역의 중추적 역할을 담당하는 T 세포의 수적 감소와 기능 저하, 단핵구와 대식세포의 침윤, 비만세포와 호산구의 수적 증가, CD4+ T 림프구 등의 수적 증가 등의 면역학적 요인이 보고되어 있으며(J Invest Dermatol., 96:523-526, 1991; J Invest Dermatol., 97:389-394, 1991; Immunol., 11:81-88, 1999; Curr Drug Targets Inflamm Allergy., 2:199-120, 2003; J Allergy Clin Immunol., 107:871-877, 2001; Adv Immunol., 78:57, 2001; International Immunology, 14(7):767-773, 2002; Pediatr Allergy Immunol., 19:605-613, 2008), 특히 Th1 세포에 비해 Th2 세포의 수적 증가로 인한 Th1/Th2 불균형이 중요한 요인으로 알려져 있다(Kor J Pharmacogn, 43:59-65, 2012). The cause of atopic dermatitis is not yet clearly defined, but an abnormal increase in IgE, a decrease in the number and function of T cells that play a central role in cellular immunity, infiltration of monocytes and macrophages, an increase in the number of mast cells and eosinophils, Immunological factors such as an increase in the number of CD4 + T lymphocytes have been reported (J Invest Dermatol., 96:523-526, 1991; J Invest Dermatol., 97:389-394, 1991; Immunol., 11:81). -88, 1999; Curr Drug Targets Inflamm Allergy., 2:199-120, 2003; J Allergy Clin Immunol., 107:871-877, 2001; Adv Immunol., 78:57, 2001; International Immunology, 14(7 ):767-773, 2002; Pediatr Allergy Immunol., 19:605-613, 2008), in particular, Th1/Th2 imbalance due to an increase in the number of Th2 cells compared to Th1 cells is known to be an important factor (Kor J Pharmacogn, 43 :59-65, 2012).
한편, 케모카인(chemokine)은 저분자 단백질로 화학주성(chemoattractants)을 나타내며 그 구조와 기능에 따라 C, CC, CXC 및 CX3C 등 4가지로 나누어지는데, 최근 이러한 케모카인들이 면역세포의 발생과 성숙, T 세포의 분화와 이주, Th1/Th2 균형 조절 등에 중요한 역할을 하는 것으로 보고 있다. On the other hand, chemokines are small-molecular proteins that exhibit chemoattractants and are divided into four types, C, CC, CXC, and CX3C, depending on their structure and function. It is believed to play an important role in the differentiation and migration of Th1/Th2 balance.
다양한 케모카인들 중 TARC(Thymus and activation-regulated chemokine)와 MDC(macrophage-derived chemokine)는 Th2 세포에 작용하는 대표적인 케모카인으로, Th2 세포의 CCR4(CC chemokine receptor 4)에 작용하여 염증 병변으로 Th2 세포의 이동과 침윤을 유도한다(J Clin Invest. 113(5):651-7, 2004; Curr Allergy Asthma Rep. 5(4):284-90, 2005)Among various chemokines, TARC (Thymus and activation-regulated chemokine) and MDC (macrophage-derived chemokine) are representative chemokines that act on Th2 cells. induce migration and invasion (J Clin Invest. 113(5):651-7, 2004; Curr Allergy Asthma Rep. 5(4):284-90, 2005)
최근 아토피 피부염 환자에서 TARC 및 MDC 인자의 혈청 농도가 현저히 증가한다는 보고가 있고(J Allergy Clin Immunol., 113(2):334-340), 아토피 피부염 동물모델에서 TARC와 MDC의 피부 발현이 증가된다는 보고가 있으며, 아토피 피부염의 치료 물질로 사용되는 사이크로스포린 A나 코르티코스테로이드를 아토피 피부염 환자에게 투여하였을 때는 MDC 및 TARC의 혈청 농도가 감소한다는 보고가 있다(J Dermatol Sci., 34:201-208, 2004). Recently, it has been reported that serum concentrations of TARC and MDC factors significantly increase in patients with atopic dermatitis (J Allergy Clin Immunol., 113(2):334-340), and that the skin expression of TARC and MDC is increased in atopic dermatitis animal models. There is a report, and there is a report that the serum concentration of MDC and TARC decreases when cyclosporin A or corticosteroid, which is used as a therapeutic substance for atopic dermatitis, is administered to atopic dermatitis patients (J Dermatol Sci., 34:201-208) , 2004).
또한, In-vitro 실험에서 인간각질형성세포주인 HaCaT 세포에 INF-γ나 TNF-α를 처리하였을 때 MDC 및 TARC가 다량 발현되는데 이러한 발현을 억제할 수 있는 물질은 아토피 피부염 치료제로 사용될 수 있음이 제시된 바 있다(Int Immunol., 14(7):767-773, 2002).In addition, in an in-vitro experiment, when INF-γ or TNF-α was treated in HaCaT cells, a human keratinocyte line, a large amount of MDC and TARC were expressed. It was found that substances capable of inhibiting these expressions can be used as a treatment for atopic dermatitis. has been suggested (Int Immunol., 14(7):767-773, 2002).
현재 아토피성 피부염 치료제로 염증 반응과 사이토카인 생산을 억제하는 스테로이드제가 주로 이용되고 있으나, 장기간 투여할 경우 피부의 위축이나 성장 지연 가능성 등 여러 가지 부작용을 초래하여 최근에는 비스테로이드제의 사용이 증가하고 있다. 그러나, 비스테로이드제 역시 홍반, 가려움, 부종, 짓무름 및 태선화 등의 증상과 면역력 약화 등의 다양한 부작용을 가지고 있으므로, 근본적인 아토피성 피부염의 치료가 어려운 실정이다(Arellano FM et al., J Invest Dermatol. 2007 Apr;127(4):808-16.). Currently, steroids that suppress the inflammatory reaction and cytokine production are mainly used as a treatment for atopic dermatitis, but when administered for a long period of time, it causes various side effects such as skin atrophy and growth retardation. have. However, since non-steroids also have various side effects such as symptoms such as erythema, itchiness, edema, erosion and lichenification, and weakened immunity, it is difficult to treat fundamental atopic dermatitis (Arellano FM et al., J Invest Dermatol. 2007 Apr;127(4):808-16.).
따라서, 아토피 치료 효과가 뛰어난 천연 물질을 발굴하고자 하는 많은 노력이 기울여졌으나, 아직까지 단일 소재로 아토피 피부염의 치료 효과가 높은 물질은 개발된 바 없으며, 새로운 시도로서 종래 아토피 치료 효과가 있다고 알려진 안전성이 높은 천연물들의 아토피 피부염 치료 효과를 높이는 방향의 연구와 같은 발상의 전환이 필요한 실정이다.Therefore, many efforts have been made to discover natural substances with excellent therapeutic effects for atopic dermatitis, but a single material with high therapeutic effects for atopic dermatitis has not yet been developed. It is a situation that requires a change of thinking, such as research in the direction of increasing the therapeutic effect of atopic dermatitis of high natural products.
이에, 본 발명자들은 아토피 피부염의 치료에 효과가 있다고 알려진 천연물질의 아토피 피부염 치료 효과를 더욱 개선시켜 스테로이드제 등의 화합물과 동등 이상의 효과를 나타낼 수 있도록 그 효과를 더욱 개선시키고자 예의 연구노력한 결과, 타임 추출물을 유산균인 락토바실러스 플란타룸 균주로 발효시킨 천연물질이 아토피 인자인 MDC 및 TARC 단백질의 생성을 효과적으로 억제하고, IL-6와 같은 염증성 사이토카인 생성을 억제하여 아토피성 면역 또는 염증반응의 억제에 매우 효과적이며, 부가적으로 MMP-1 단백질 생성 억제 및 타입 1 프로콜라겐 단백질 생성 증진을 통해 피부 주름의 예방 및 개선에도 효과적임을 확인하여 종래 부작용이 많아 사용이 어려운 아토피 피부염 치료용 화합물을 대체하여 사용할 수 있음을 확인하고, 본 발명을 완성하게 되었다.Accordingly, the present inventors further improved the therapeutic effect of atopic dermatitis of natural substances known to be effective in the treatment of atopic dermatitis to further improve the effect so that it can exhibit an effect equal to or greater than that of a compound such as a steroid. A natural substance fermented with thyme extract with a Lactobacillus plantarum strain, a lactic acid bacterium, effectively inhibits the production of MDC and TARC proteins, which are atopic factors, and inhibits the production of inflammatory cytokines such as IL-6 to improve atopic immunity or inflammatory response. It is very effective in suppression, and additionally, it is effective in preventing and improving skin wrinkles through inhibition of MMP-1 protein production and promotion of
따라서, 본 발명의 주된 목적은 아토피 피부염 및 피부주름 개선에 우수한 효과를 갖는 유산균 발효 타임 추출물을 유효성분으로 함유하는 아토피 피부염 및 피부주름 개선용 조성물을 제공하는 데 있다.Therefore, the main object of the present invention is to provide a composition for improving atopic dermatitis and skin wrinkles containing as an active ingredient a lactic acid bacterium fermented thyme extract having an excellent effect on improving atopic dermatitis and skin wrinkles.
본 발명의 다른 목적 및 이점은 하기의 발명의 상세한 설명, 청구범위 및 도면에 의해 보다 명확하게 된다.Other objects and advantages of the present invention will become more apparent from the following detailed description of the invention, claims and drawings.
본 발명의 한 양태에 따르면, 본 발명은 락토바실러스 플란타룸(기탁번호: KCTC 14013BP) 발효 타임 추출물을 유효성분으로 함유하는 아토피 피부염 및 피부주름 개선용 화장료 조성물을 제공한다.According to one aspect of the present invention, the present invention provides a cosmetic composition for improving atopic dermatitis and skin wrinkles containing Lactobacillus plantarum (Accession No.: KCTC 14013BP) fermented thyme extract as an active ingredient.
본 발명자들은 아토피 피부염의 치료에 효과가 있다고 알려진 천연물질의 아토피 피부염 치료 효과를 더욱 개선시켜 스테로이드제 등의 화합물과 동등 이상의 효과를 나타낼 수 있도록 천연물 발효를 통해 그 효과를 더욱 개선하고자 예의 연구노력하였고, 유산균으로 발효한 타임 발효추출물의 아토피 피부염 개선 효과가 우수함을 확인하였다. The present inventors have worked diligently to further improve the effect of natural substances known to be effective in the treatment of atopic dermatitis through fermentation of natural products so as to further improve the therapeutic effect of atopic dermatitis, thereby exhibiting an effect equal to or greater than that of compounds such as steroids. , it was confirmed that the thyme fermented extract fermented with lactic acid bacteria was excellent in the improvement of atopic dermatitis.
본 발명자들은 상기 타임 발효에 최적화된 유산균의 특성을 종합하여 조사한 결과, 타임 발효에 가장 최적화된 효율을 내는 균주가 락토바실러스 플란타룸 균주임을 확인하였고, 이를 “락토바실러스 플란타룸 아종 플란타룸(Lactobacillus plantarum subsp. plantarum) SDCM1002”로 명명하고, 한국생명공학연구원 생물자원센터에 2019년 11월 4일자로 기탁번호 KCTC 14013BP로 기탁하였다. 상기 본 발명의 락토바실러스 플란타룸 아종 플란타룸 SDCM1002(기탁번호: KCTC 14013BP) 균주는 락토바실러스 플란타룸(기탁번호: KCTC 14013BP)와 동일한 의미로 사용된다. As a result of investigating the characteristics of lactic acid bacteria optimized for the thyme fermentation, the present inventors confirmed that the strain with the most optimal efficiency for thyme fermentation was the Lactobacillus plantarum strain, which ( Lactobacillus plantarum subsp. plantarum ) SDCM1002”, and deposited with the Korea Research Institute of Bioscience and Biotechnology Biological Resources Center as of November 4, 2019 with deposit number KCTC 14013BP. The Lactobacillus plantarum subspecies plantarum SDCM1002 (accession number: KCTC 14013BP) strain of the present invention is used in the same meaning as the Lactobacillus plantarum (accession number: KCTC 14013BP).
본 발명의 "타임(Thymus vulgaris)"은 쌍떡잎식물 통화식물목 꿀풀과의 여러해살이풀을 의미한다. "Thyme (Thymus vulgaris)" of the present invention means a dicotyledonous perennial plant of the order Lamiaceae.
본 발명에서 용어, "아토피 피부염"은 만성적이고 재발성의 염증성 피부질환으로 가려움증(가려움증)과 피부건조증, 특징적인 홍반 또는 습진을 동반하는 질병을 의미한다. 아토피성 피부염의 급성 병소에서는 혈청 면역글로불린 E(IgE)의 유의한 증가 현상이 특징으로 나타나며, 이에 부가하여 병소의 조직병리학적인 변화 및 피부염 병변에 대한 평가 등을 행하여 아토피성 피부염의 진단 및 심각도를 판정하기도 한다. 아토피성 피부염의 정확한 원인은 아직까지 완전히 이해되고 있지 않지만 유전적 소인과 함께 면역학적, 비면역학적 기전이 관여한다고 보고 있다.As used herein, the term "atopic dermatitis" refers to a chronic and recurrent inflammatory skin disease accompanied by itching (itchiness), dry skin, and characteristic erythema or eczema. Acute lesions of atopic dermatitis are characterized by a significant increase in serum immunoglobulin E (IgE). also judge. Although the exact cause of atopic dermatitis is not yet fully understood, it is believed that immunological and non-immunological mechanisms are involved along with genetic predisposition.
본 발명에서 용어, "피부 주름"이란, 피부가 쇠하여 생긴 잔줄을 의미하는데, 유전자에 의한 원인, 피부 진피에 존재하는 콜라겐의 감소, 외부 환경 등에 의해 유발될 수 있다.As used herein, the term "skin wrinkle" refers to fine lines caused by skin aging, and may be caused by genetic causes, a decrease in collagen present in the dermis of the skin, external environment, and the like.
또한, 본 명세서에서 "유효성분"이란 단독으로 목적하는 활성을 나타내거나, 또는 그 자체는 활성이 없는 담체와 함께 활성을 나타낼 수 있는 성분을 의미한다.In addition, as used herein, the term "active ingredient" refers to a component that alone exhibits a desired activity, or can exhibit activity together with a carrier that has no activity by itself.
본 발명에서 사용되는 용어, "추출물"은 식물의 추출 처리로 얻어지는 추출액, 상기 추출액의 희석액이나 농축액, 상기 추출액을 건조하여 얻어지는 건조물, 상기 추출액의 조정제물이나 정제물, 또는 이들의 혼합물 등, 추출액 자체 및 추출액을 이용하여 형성 가능한 모든 제형의 추출물을 포함한다. 본 발명의 상기 추출물은 바람직하게 추출 후 건조 분말 형태로 제조되어 사용될 수 있다. As used in the present invention, the term "extract" refers to an extract obtained by extraction of plants, a diluted or concentrated liquid of the extract, a dried product obtained by drying the extract, a prepared or purified product of the extract, or a mixture thereof, such as an extract It includes extracts of all formulations that can be formed by themselves and using extracts. The extract of the present invention may be prepared and used in the form of a dry powder after extraction.
본 발명의 상기 타임 추출물의 추출에 있어서, 상기 추출물을 추출하는 방법은 특별히 제한되지 아니하며, 당해 기술 분야에서 통상적으로 사용하는 방법에 따라 추출할 수 있다. 상기 추출방법의 비제한적인 예로는, 열수 추출법, 초음파 추출법, 여과법, 환류 추출법 등을 들 수 있으며, 이들은 단독으로 수행되거나 2종 이상의 추출방법을 병용하여 수행될 수 있다. In the extraction of the thyme extract of the present invention, the method of extracting the extract is not particularly limited, and may be extracted according to a method commonly used in the art. Non-limiting examples of the extraction method include hot water extraction, ultrasonic extraction, filtration, reflux extraction, and the like, and these may be performed alone or in combination of two or more extraction methods.
본 발명에서 상기 타임을 추출하는데 사용되는 추출 용매의 종류는 특별히 제한되지 아니하며, 당해 기술 분야에서 공지된 임의의 용매를 사용할 수 있다. 상기 추출 용매의 비제한적인 예로는 물; 메탄올, 에탄올, 프로필알코올, 부틸알코올 등의 C1 내지 C4의 저급 알코올; 글리세린, 부틸렌글리콜, 프로필렌글리콜 등의 다가 알코올; 및 메틸아세테이트, 에틸아세테이트, 아세톤, 벤젠, 헥산, 디에틸에테르, 디클로로메탄 등의 탄화수소계 용매; 또는 이들의 혼합물을 사용할 수 있으며, 바람직하게, 물, 저급알코올, 1,3-부틸렌글리콜, 에틸아세테이트를 단독으로 사용하거나 2종 이상 혼합하여 사용할 수 있다. In the present invention, the type of the extraction solvent used for extracting the thyme is not particularly limited, and any solvent known in the art may be used. Non-limiting examples of the extraction solvent include water; C1-C4 lower alcohols, such as methanol, ethanol, propyl alcohol, and butyl alcohol; polyhydric alcohols such as glycerin, butylene glycol and propylene glycol; and hydrocarbon solvents such as methyl acetate, ethyl acetate, acetone, benzene, hexane, diethyl ether, and dichloromethane; Alternatively, a mixture thereof may be used, and preferably, water, lower alcohol, 1,3-butylene glycol, and ethyl acetate may be used alone or in a mixture of two or more.
본 발명에서 열수 추출 또는 냉침 추출한 추출물은 부유하는 고체 입자를 제거하기 위해 여과, 예를 들어 나일론 등을 이용해 입자를 걸러내거나 냉동여과법 등을 이용해 여과한 후, 그대로 사용하거나 이를 동결건조, 열풍건조, 분무건조 등을 이용해 건조하여 사용할 수 있다.In the present invention, the extract extracted with hot water or chilled extract is filtered to remove floating solid particles, for example, using nylon or the like to filter the particles, or after filtration using a freeze filtration method, etc. It can be used by drying it using spray drying or the like.
본 발명의 화장료 조성물은 TARC(Thymus and activation-regulated chemokine) 및 MDC(macrophage-derived chemokine) 생성을 억제하는 것을 특징으로 하며(도 2 및 도 3 참조), IL-6와 같은 염증성 사이토카인 생성을 억제하여(도 4 참조), 아토피성 면역 또는 염증반응의 억제에 매우 효과적이다. The cosmetic composition of the present invention is characterized in that it inhibits TARC (Thymus and activation-regulated chemokine) and MDC (macrophage-derived chemokine) production (see FIGS. 2 and 3 ), and inhibits the production of inflammatory cytokines such as IL-6 By suppressing (see FIG. 4), it is very effective in suppressing atopic immunity or inflammatory response.
또한, 본 발명의 화장료 조성물은 MMP-1 단백질 생성을 억제하고, 타입 1 프로콜라겐 단백질 생성을 증가시키는 것을 특징으로 한다(도 5 및 도 6 참조). In addition, the cosmetic composition of the present invention is characterized in that it inhibits the production of MMP-1 protein and increases the production of
본 발명의 아토피 피부염 및 피부주름 개선용 화장료 조성물에서, 상기 조성물은 용액, 외용연고, 크림, 폼, 영양화장수, 유연화장수, 마스크 팩, 유연수, 유액, 메이크업베이스, 에센스, 비누, 액체 세정료, 입욕제, 선 스크린크림, 선오일, 현탁액, 유탁액, 페이스트, 겔, 로션, 파우더, 비누, 계면활성제-함유 클린싱, 오일, 분말 파운데이션, 유탁액 파운데이션, 왁스 파운데이션, 패치 및 스프레이로 구성된 군으로부터 선택되는 제형으로 제조할 수 있으나, 이에 제한되는 것은 아니다.In the cosmetic composition for improving atopic dermatitis and skin wrinkles of the present invention, the composition comprises a solution, an external ointment, a cream, a foam, a nourishing lotion, a softening lotion, a mask pack, a softening water, an emulsion, a makeup base, an essence, a soap, a liquid detergent, Select from the group consisting of bath products, sunscreen creams, sun oils, suspensions, emulsions, pastes, gels, lotions, powders, soaps, surfactant-containing cleansers, oils, powder foundations, emulsion foundations, wax foundations, patches and sprays It can be prepared in a dosage form, but is not limited thereto.
또한, 본 발명의 화장료 조성물은 일반 피부 화장료에 배합되는 화장품학적으로 허용 가능한 담체를 1종 이상 추가로 포함할 수 있으며, 통상의 성분으로 예를 들면 유분, 물, 계면활성제, 보습제, 저급 알콜, 증점제, 킬레이트제, 색소, 방부제, 향료 등을 적절히 배합할 수 있으나, 이에 제한되는 것은 아니다.In addition, the cosmetic composition of the present invention may further include one or more cosmetically acceptable carriers to be formulated in general skin cosmetics, and conventional ingredients include, for example, oil, water, surfactant, humectant, lower alcohol, A thickener, a chelating agent, a colorant, a preservative, a fragrance, etc. may be appropriately mixed, but the present invention is not limited thereto.
본 발명의 화장료 조성물에 포함되는 화장품학적으로 허용 가능한 담체는 제형에 따라 다양하다.The cosmetically acceptable carrier included in the cosmetic composition of the present invention varies depending on the formulation.
본 발명의 제형이 연고, 페이스트, 크림 또는 젤인 경우에는, 담체 성분으로서 동물성 유, 식물성 유, 왁스, 파라핀, 전분, 트라칸트, 셀룰로오스 유도체, 폴리에틸렌 글리콜, 실리콘, 벤토나이트, 실리카, 탈크, 산화아연 또는 이들의 혼합물이 이용될 수 있다.When the formulation of the present invention is an ointment, paste, cream or gel, animal oil, vegetable oil, wax, paraffin, starch, tracanth, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc, zinc oxide or Mixtures thereof may be used.
본 발명의 제형이 파우더 또는 스프레이인 경우에는, 담체 성분으로서 락토스, 탈크, 실리카, 알루미늄 히드록사이드, 칼슘 실케이트, 폴리아미드 파우더 또는 이들의 혼합물이 이용될 수 있고, 특히 스프레이인 경우에는 추가적으로 클로로플루오로히드로카본, 프로판/부탄 또는 디메틸에테르와 같은 추진제를 포함할 수 있다.When the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate, polyamide powder, or mixtures thereof may be used as a carrier component. In particular, in the case of a spray, additional chlorophyll It may contain propellants such as fluorohydrocarbons, propane/butane or dimethyl ether.
본 발명의 제형이 용액 또는 유탁액인 경우에는, 담체 성분으로서 용매, 용해화제 또는 유탁화제가 이용되며, 예컨대 물, 에탄올, 이소프로판올, 에틸 카보네이트, 에틸 아세테이트, 벤질 알콜, 벤질 벤조에이트, 프로필렌글리콜, 1,3-부틸글리콜 오일이 이용될 수 있으며, 특히, 목화씨 오일, 땅콩 오일, 옥수수 배종 오일, 올리브오일, 피마자 오일 및 참깨 오일, 글리세롤 지방족 에스테르, 폴리에틸렌 글리콜 또는 소르비탄의 지방산 에스테르가 이용될 수 있다.When the formulation of the present invention is a solution or emulsion, a solvent, solubilizer or emulsifier is used as a carrier component, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylglycol oil may be used, and in particular, cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil, glycerol fatty ester, fatty acid ester of polyethylene glycol or sorbitan may be used. have.
본 발명의 제형이 현탁액인 경우에는, 담체 성분으로서 물, 에탄올 또는 프로필렌 글리콜과 같은 액상의 희석제, 에톡실화 이소스테아릴 알콜, 폴리옥시에틸렌 소르비톨 에스테르 및 폴리옥시에틸렌 소르비탄 에스테르와 같은 현탁제, 미소결정성 셀룰로오스, 알루미늄 메타히드록시드, 벤토나이트, 아가 또는 트라칸트 등이 이용될 수 있다.When the formulation of the present invention is a suspension, as a carrier component a liquid diluent such as water, ethanol or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol esters and polyoxyethylene sorbitan esters; Crystalline cellulose, aluminum metahydroxide, bentonite, agar or tracanth may be used.
본 발명의 제형이 비누인 경우에는 담체 성분으로서 지방산의 알칼리 금속 염, 지방산 헤미에스테르 염, 지방산 단백질 히드롤리제이트, 이세티오네이트, 라놀린 유도체, 지방족 알콜, 식물성 유, 글리세롤, 당 등이 이용될 수 있다.When the formulation of the present invention is soap, alkali metal salts of fatty acids, fatty acid hemiester salts, fatty acid protein hydrolysates, isethionate, lanolin derivatives, aliphatic alcohols, vegetable oils, glycerol, sugar, etc. may be used as carrier components. can
본 발명의 다른 양태에 따르면, 본 발명은 락토바실러스 플란타룸(기탁번호: KCTC 14013BP) 발효 타임 추출물을 유효성분으로 함유하는 아토피 피부염 및 피부주름 개선용 식품 조성물을 제공한다. 상기 락토바실러스 플란타룸(기탁번호: KCTC 14013BP) 균주와 아토피 피부염 및 피부주름 개선은 상기 화장료 조성물에서 설명한 바와 같다. According to another aspect of the present invention, the present invention provides a food composition for improving atopic dermatitis and skin wrinkles containing Lactobacillus plantarum (Accession No.: KCTC 14013BP) fermented thyme extract as an active ingredient. The Lactobacillus plantarum (Accession No.: KCTC 14013BP) strain and the improvement of atopic dermatitis and skin wrinkles are the same as those described in the cosmetic composition.
본 발명의 식품 조성물은 TARC(Thymus and activation-regulated chemokine) 및 MDC(macrophage-derived chemokine) 생성을 억제하고, MMP-1 단백질 생성을 억제하며, 타입 1 프로콜라겐 단백질 생성을 증가시키는 우수한 효과가 있어 아토피 피부염 및 피부주름 개선용 식품 조성물로 유용하게 사용될 수 있다. 상기 식품 조성물은 건강기능식품의 형태로 사용될 수 있으나, 이에 제한되는 것은 아니다. The food composition of the present invention has an excellent effect of inhibiting TARC (Thymus and activation-regulated chemokine) and MDC (macrophage-derived chemokine) production, inhibiting MMP-1 protein production, and increasing
본 발명의 식품 조성물은 락토바실러스 플란타룸(기탁번호: KCTC 14013BP) 발효 타임 추출물의 분획물 또는 이의 가공물의 형태로 포함될 수 있다. 또한, 상기 조성물은 유효성분 이외에 식품학적으로 허용 가능한 식품보조첨가제를 포함할 수 있다.The food composition of the present invention may be included in the form of a fraction of Lactobacillus plantarum (Accession No.: KCTC 14013BP) fermentation thyme extract or a processed product thereof. In addition, the composition may include a food pharmaceutically acceptable food supplement additive in addition to the active ingredient.
본 발명에 있어서, "식품보조첨가제"란 식품에 보조적으로 첨가될 수 있는 구성요소를 의미하며, 각 제형의 건강기능식품을 제조하는데 첨가되는 것으로서 당업자가 적절히 선택하여 사용할 수 있다. 식품보조첨가제의 예로는 여러 가지 영양제, 비타민, 광물(전해질), 합성 풍미제 및 천연 풍미제 등의 풍미제, 착색제 및 충진제, 펙트산 및 그의 염, 알긴산 및 그의 염, 유기산, 보호성 콜로이드 증점제, pH 조절제, 안정화제, 방부제, 글리세린, 알콜, 탄산음료에 사용되는 탄산화제 등이 포함되지만, 상기 예들에 의해 본 발명의 식품보조첨가제의 종류가 제한되는 것은 아니다.In the present invention, "food supplement additive" refers to a component that can be added to food as an auxiliary, added to the manufacture of health functional food of each formulation, and can be appropriately selected and used by those skilled in the art. Examples of food supplement additives include various nutrients, vitamins, minerals (electrolytes), synthetic flavoring agents and flavoring agents such as natural flavoring agents, coloring agents and fillers, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners , pH adjuster, stabilizer, preservative, glycerin, alcohol, carbonation agent used in carbonated beverages, etc., but the above examples are not limited to the type of food supplement additive of the present invention.
본 발명의 식품 조성물에는 건강기능식품이 포함될 수 있다. 본 발명에 있어서, "건강기능식품"이란 인체에 유용한 기능성을 가진 원료나 성분을 사용하여 정제, 캅셀, 분말, 과립, 액상 및 환 등의 형태로 제조 및 가공한 식품을 말한다. 여기서 “기능성”이라 함은 인체의 구조 및 기능에 대하여 영양소를 조절하거나 생리학적 작용 등과 같은 보건 용도에 유용한 효과를 얻는 것을 의미한다. 본 발명의 건강기능식품은 당업계에서 통상적으로 사용되는 방법으로 제조 가능하며, 상기 제조시에는 당업계에서 통상적으로 첨가하는 원료 및 성분을 첨가하여 제조할 수 있다. The food composition of the present invention may include a health functional food. In the present invention, "health functional food" refers to food manufactured and processed in the form of tablets, capsules, powders, granules, liquids, pills, etc. using raw materials or ingredients useful for the human body. Here, the term “functionality” refers to obtaining useful effects for health purposes, such as regulating nutrients or physiological actions with respect to the structure and function of the human body. The health functional food of the present invention can be prepared by a method commonly used in the art, and at the time of manufacture, it can be prepared by adding raw materials and components commonly added in the art.
또한, 상기 건강기능식품의 제형 또한 건강기능식품으로 인정되는 제형이면 제한 없이 제조될 수 있다. 본 발명의 식품 조성물은 다양한 형태의 제형으로 제조될 수 있으며, 일반 약품과는 달리 식품을 원료로 하여 약품의 장기 복용 시 발생할 수 있는 부작용 등이 없는 장점이 있고, 휴대성이 뛰어나, 아토피 피부염 및 피부 주름을 개선하는 효과를 증진시키기 위한 보조제로 섭취가 가능하다.In addition, if the formulation of the health functional food is also recognized as a health functional food, it may be manufactured without limitation. The food composition of the present invention can be prepared in various types of dosage forms, and unlike general drugs, it uses food as a raw material and does not have side effects that may occur when taking the drug for a long period of time, and has excellent portability, so that atopic dermatitis and It can be taken as a supplement to enhance the effect of improving skin wrinkles.
본 발명의 건강기능식품이 취할 수 있는 형태에는 제한이 없으며, 통상적인 의미의 식품을 모두 포함할 수 있고, 기능성 식품 등 당업계에 알려진 용어와 혼용이 가능하다. 아울러 본 발명의 건강기능식품은 당업자의 선택에 따라 식품에 포함될 수 있는 적절한 기타 보조 성분과 공지의 첨가제를 혼합하여 제조할 수 있다. 첨가할 수 있는 식품의 예로는 육류, 소세지, 빵, 초콜릿, 캔디류, 스낵류, 과자류, 피자, 라면, 기타 면류, 껌류, 아이스크림류를 포함한 낙농제품, 각종 스프, 음료수, 차, 드링크제, 알코올 음료 및 비타민 복합제 등이 있으며, 본 발명에 따른 화학식 1로 표시되는 화합물을 주성분으로 하여 제조한 즙, 차, 젤리 및 주스 등에 첨가하여 제조할 수 있다. 또한, 동물을 위한 사료로 이용되는 식품도 포함된다.There is no limitation in the form that the health functional food of the present invention can take, and it may include all foods in a conventional sense, and may be used interchangeably with terms known in the art, such as functional food. In addition, the health functional food of the present invention can be prepared by mixing known additives with other suitable auxiliary ingredients that may be included in food according to the selection of those skilled in the art. Examples of foods that can be added include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, dairy products including ice cream, various soups, beverages, tea, drinks, alcoholic beverages, and There are vitamin complexes, and the like, and it can be prepared by adding the compound represented by
본 발명의 다른 양태에 따르면, 본 발명은 락토바실러스 플란타룸(기탁번호: KCTC 14013BP) 발효 타임 추출물을 유효성분으로 함유하는 아토피 피부염 치료 또는 예방용 약학적 조성물을 제공한다. 상기 락토바실러스 플란타룸(기탁번호: KCTC 14013BP) 균주와 아토피 피부염 및 피부주름 개선은 상기 화장료 조성물에서 설명한 바와 같다. According to another aspect of the present invention, the present invention provides a pharmaceutical composition for treating or preventing atopic dermatitis containing Lactobacillus plantarum (Accession No.: KCTC 14013BP) fermented thyme extract as an active ingredient. The Lactobacillus plantarum (Accession No.: KCTC 14013BP) strain and the improvement of atopic dermatitis and skin wrinkles are the same as those described in the cosmetic composition.
본 발명의 약학적 조성물은 TARC(Thymus and activation-regulated chemokine) 및 MDC(macrophage-derived chemokine) 생성을 억제하고, IL-6 단백질 생성을 억제하는 우수한 효과가 있어 아토피 피부염 치료 또는 예방용 약학적 조성물로 유용하게 사용될 수 있다. The pharmaceutical composition of the present invention has an excellent effect of inhibiting TARC (Thymus and activation-regulated chemokine) and MDC (macrophage-derived chemokine) production, and inhibiting IL-6 protein production, so a pharmaceutical composition for treating or preventing atopic dermatitis can be usefully used as
본 발명의 용어 "치료"는 본 발명에 따른 조성물의 투여로 상기 아토피 피부염 질환의 증세가 호전되거나 이롭게 변경되는 모든 행위를 의미하고, 용어 "예방"은 본 발명에 따른 조성물의 투여로 상기 아토피 피부염 관련 질환의 발병을 억제 또는 지연시키는 모든 행위를 의미한다.The term "treatment" of the present invention refers to any action in which the symptoms of the atopic dermatitis disease are improved or beneficially changed by administration of the composition according to the present invention, and the term "prevention" is the atopic dermatitis by administration of the composition according to the present invention It means any action that suppresses or delays the onset of related diseases.
본 발명의 조성물이 약제학적 조성물로 제조되는 경우, 본 발명의 약제학적 조성물은 약제학적으로 허용되는 담체를 포함한다. 본 발명의 약제학적 조성물에 포함되는 약제학적으로 허용되는 담체는 제제시에 통상적으로 이용되는 것으로서, 락토스, 덱스트로스, 수크로스, 솔비톨, 만니톨, 전분, 아카시아 고무, 인산 칼슘, 알기네이트, 젤라틴, 규산 칼슘, 미세결정성 셀룰로스, 폴리비닐피롤리돈, 셀룰로스, 물, 시럽, 메틸 셀룰로스, 메틸히드록시벤조에이트, 프로필히드록시벤조에이트, 활석, 스테아르산 마그네슘 및 미네랄 오일 등을 포함하나, 이에 한정되는 것은 아니다. 본 발명의 약제학적 조성물은 상기 성분들 이외에 윤활제, 습윤제, 감미제, 향미제, 유화제, 현탁제, 보존제 등을 추가로 포함할 수 있다. 적합한 약제학적으로 허용되는 담체 및 제제는 Remington's Pharmaceutical Sciences(19th ed., 1995)에 상세히 기재되어 있다.When the composition of the present invention is prepared as a pharmaceutical composition, the pharmaceutical composition of the present invention includes a pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers included in the pharmaceutical composition of the present invention are commonly used in formulation, and include lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia gum, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil; it is not going to be The pharmaceutical composition of the present invention may further include a lubricant, a wetting agent, a sweetening agent, a flavoring agent, an emulsifying agent, a suspending agent, a preservative, and the like, in addition to the above components. Suitable pharmaceutically acceptable carriers and agents are described in detail in Remington's Pharmaceutical Sciences (19th ed., 1995).
본 발명의 약제학적 조성물은 경구 또는 비경구 투여할 수 있으며, 본 발명의 일 구현예에 따르면 비경구 방식으로 투여되고, 본 발명의 다른 구현예에 따르면 경피투여방식으로 투여된다. The pharmaceutical composition of the present invention may be administered orally or parenterally. According to one embodiment of the present invention, it is administered by a parenteral method, and according to another embodiment of the present invention, it is administered by a transdermal administration method.
본 발명의 약제학적 조성물의 적합한 투여량은 제제화 방법, 투여 방식, 환자의 연령, 체중, 성, 병적 상태, 음식, 투여 시간, 투여 경로, 배설 속도 및 반응 감응성과 같은 요인들에 의해 다양하게 처방될 수 있다. 본 발명의 약제학적 조성물의 바람직한 투여량은 성인 기준으로 0.0001-100㎎/kg 범위 내이다.A suitable dosage of the pharmaceutical composition of the present invention is variously prescribed depending on factors such as formulation method, administration method, age, weight, sex, pathological condition, food, administration time, administration route, excretion rate, and reaction sensitivity of the patient. can be A preferred dosage of the pharmaceutical composition of the present invention is within the range of 0.0001-100 mg/kg for adults.
본 발명의 약제학적 조성물은 당해 발명이 속하는 기술분야에서 통상의 지식을 가진 자가 용이하게 실시할 수 있는 방법에 따라, 약제학적으로 허용되는 담체 및/또는 부형제를 이용하여 제제화함으로써 단위 용량 형태로 제조되거나 또는 다용량 용기 내에 내입시켜 제조될 수 있다. 이때 제형은 오일 또는 수성 매질중의 용액, 현탁액, 시럽제 또는 유화액 형태이거나 엑스제, 산제, 분말제, 과립제, 정제 또는 캅셀제 형태일 수도 있으며, 분산제 또는 안정화제를 추가적으로 포함할 수 있다.The pharmaceutical composition of the present invention is prepared in unit dosage form by formulating using a pharmaceutically acceptable carrier and/or excipient according to a method that can be easily performed by a person of ordinary skill in the art to which the present invention pertains. or it may be prepared by incorporation into a multi-dose container. In this case, the formulation may be in the form of a solution, suspension, syrup, or emulsion in oil or an aqueous medium, or may be in the form of an extract, powder, powder, granule, tablet or capsule, and may additionally include a dispersant or stabilizer.
본 발명의 특징 및 이점을 요약하면 다음과 같다:The features and advantages of the present invention are summarized as follows:
(a) 본 발명은 락토바실러스 플란타룸(기탁번호: KCTC 14013BP) 발효 타임 추출물을 유효성분으로 함유하는 아토피 피부염 및 피부주름 개선용 조성물을 제공한다.(a) The present invention provides a composition for improving atopic dermatitis and skin wrinkles containing Lactobacillus plantarum (Accession No.: KCTC 14013BP) fermented thyme extract as an active ingredient.
(b) 본 발명의 유산균 발효 타임 추출물은 아토피 인자인 TARC 및 MDC 단백질의 생성을 효과적으로 억제하고, IL-6와 같은 염증성 사이토카인 생성을 억제하여 아토피성 면역 또는 염증 반응의 억제에 매우 효과적이며, MMP-1 단백질 생성 억제 및 타입 1 프로콜라겐 단백질 생성 증진을 통해 피부 주름의 예방 및 개선에도 효과적이므로, 아토피 피부염 및 피부주름 개선을 위한 화장료, 식품, 및 약학적 조성물로 유용하게 이용될 수 있다. (b) the lactic acid bacteria fermented thyme extract of the present invention effectively inhibits the production of TARC and MDC proteins, which are atopic factors, and inhibits the production of inflammatory cytokines such as IL-6. Since it is effective in preventing and improving skin wrinkles through inhibition of MMP-1 protein production and promotion of
(c) 또한, 유산균 발효 타임 추출물은 천연물인 타임 추출물을 유산균으로 발효한 발효추출물로 인체에 매우 안전할 뿐만 아니라, 안정성도 매우 탁월하다.(c) In addition, lactic acid bacteria fermented thyme extract is a fermented extract obtained by fermenting thyme extract, a natural product, with lactic acid bacteria, which is not only very safe for the human body, but also has excellent stability.
도 1은 본 발명의 유산균 발효 타임 추출물의 세포독성을 측정하여 나타낸 그래프이다.
도 2는 유산균 발효 타임 추출물이 처리된 인간각질형성세포(HaCaT, Human immortalized keratinocyte)에서 TARC의 발현 변화를 측정한 나타내는 그래프이다.
도 3은 유산균 발효 타임 추출물이 처리된인간각질형성세포(HaCaT, Human immortalized keratinocyte)에서 MDC의 발현 변화를 측정한 나타내는 그래프이다.
도 4는 유산균 발효 타임 추출물이 처리된 인간각질형성세포(HaCaT, Human immortalized keratinocyte)에서 IL-6의 발현 변화를 측정한 나타내는 그래프이다.
도 5는 유산균 발효 타임 추출물이 처리된 인간섬유아세포에서 MMP-1 단백질의 발현 변화를 측정한 나타내는 그래프이다.
도 6은 유산균 발효 타임 추출물이 처리된 인간섬유아세포에서 타입 1 프로콜라겐 단백질의 발현 변화를 측정한 나타내는 그래프이다. 1 is a graph showing the measurement of the cytotoxicity of the lactic acid bacteria fermentation thyme extract of the present invention.
Figure 2 is a graph showing the measurement of the expression change of TARC in human keratinocytes (HaCaT, Human immortalized keratinocytes) treated with lactic acid bacteria fermentation thyme extract.
3 is a graph showing the measurement of the expression change of MDC in human keratinocytes (HaCaT, Human immortalized keratinocyte) treated with lactic acid bacteria fermentation thyme extract.
Figure 4 is a graph showing the measurement of the expression change of IL-6 in human keratinocytes (HaCaT, Human immortalized keratinocyte) treated with lactic acid bacteria fermentation thyme extract.
5 is a graph showing the measurement of the expression change of MMP-1 protein in human fibroblasts treated with lactic acid bacteria fermentation thyme extract.
6 is a graph showing changes in the expression of
이하, 실시예를 통하여 본 발명을 더욱 상세히 설명하기로 한다. 이들 실시예는 단지 본 발명을 예시하기 위한 것이므로, 본 발명의 범위가 이들 실시예에 의해 제한되는 것으로 해석되지는 않는다.Hereinafter, the present invention will be described in more detail through examples. These examples are only for illustrating the present invention, and therefore, the scope of the present invention is not to be construed as being limited by these examples.
실시예 1. 유산균 발효 타임 추출물의 제조Example 1. Preparation of lactic acid bacteria fermentation thyme extract
타임을 흐르는 물에 세척하여 불순물을 제거하였다. 타임 600g에 정제수 6L를 첨가한 후, 3일간 실온에서 교반하여 추출하였다. 상기 과정은 3-Batch로 진행하였다. 얻어진 추출액을 모은 후, 40℃ 이하에서 감압 농축하여 엑기스 형태의 농축 추출물을 얻었다. 농축한 타임 추출물을 정제수 10mL에 1mg 비율로 녹인 후, 락토바실러스 플란타룸(Lactobacilus plantarum) 균주를 접종하여 55℃ 배양기에서 24시간 배양하였다. The thyme was washed with running water to remove impurities. After adding 6 L of purified water to 600 g of thyme, the mixture was extracted by stirring at room temperature for 3 days. The process was carried out in 3-Batch. After collecting the obtained extract, it was concentrated under reduced pressure at 40° C. or less to obtain a concentrated extract in the form of an extract. After the concentrated thyme extract was dissolved in 10 mL of purified water at a ratio of 1 mg, the Lactobacillus plantarum strain was inoculated and cultured for 24 hours in an incubator at 55°C.
배양물을 121℃에서 15분간 멸균하여 균주를 불활성화 시킨 뒤 원심분리하여 상층액만을 모았다. 이렇게 얻은 유산균 발효 타임 추출물을 감압농축기(Rotary evaporator N-1000, EYELA, Japan)를 이용하여 55℃에서 감압 농축한 후, 동결건조하여 분말형태로 제조하였다.The culture was sterilized at 121° C. for 15 minutes to inactivate the strain, and then centrifuged to collect only the supernatant. The lactic acid bacteria fermentation thyme extract thus obtained was concentrated under reduced pressure at 55° C. using a vacuum concentrator (Rotary evaporator N-1000, EYELA, Japan), and then freeze-dried to prepare a powder.
비교실험을 위해서, 유산균을 처리하여 발효하는 공정을 제외하고 상기와 같은 방법으로 유산균 발효처리 하지 않은 타임 추출물을 비교예로 사용하였다. For the comparative experiment, a thyme extract that was not fermented by lactic acid bacteria in the same manner as above except for the process of fermenting by treating lactic acid bacteria was used as a comparative example.
실험예 1. 세포독성 시험Experimental Example 1. Cytotoxicity test
상기 실시예 1에서 제조한 유산균 발효 타임 추출물이 인간각질형성세포(HaCaT, Human immortalized keratinocyte)의 생장에 미치는 효과를 MTT 비색정량분석법(colorimetric assay)을 사용하여 확인하였다. The effect of the lactic acid bacteria fermented thyme extract prepared in Example 1 on the growth of human immortalized keratinocytes (HaCaT) was confirmed using MTT colorimetric assay.
우선, 10% FBS(fetal bovine serum, Cambrex)가 포함된 전용 배지인 DMEM(Dulbeccos modified Eagles medium) 배지를 사용하여 24 well 세포 배양 접시에 인간각질형성세포를 1×105 농도로 접종한 후, 24시간 동안 37℃, 5% CO2 습윤 조건에서 배양하였다. 이후, 배지를 제거하고 무혈청 DMEM 배지로 희석한 실시예 1에서 제조한 유산균 발효 타임 추출물을 농도별(10, 및 100㎍/mL)로 처리하여 24시간 동안 배양하였다. 24시간 후, 1mg/mL의 MTT 시약을 처리하고 다시 2시간 후에 MTT 처리로 인해 세포 내에 생성된 포마잔(formazan)을 DMSO로 녹여 570nm에서 흡광도를 측정하였다. 양성대조군으로 타크롤리무스(tacrolimus)(1 및 10㎍/mL)를 처리하였다.First, using DMEM (Dulbeccos modified Eagles medium) medium, which is a dedicated medium containing 10% FBS (fetal bovine serum, Cambrex), inoculated human keratinocytes at a concentration of 1×10 5 in a 24-well cell culture dish, Incubated for 24 hours at 37° C., 5% CO 2 humid conditions. Thereafter, the medium was removed and the lactic acid bacteria fermentation thyme extract prepared in Example 1 diluted with serum-free DMEM medium was treated with each concentration (10, and 100 μg/mL) and cultured for 24 hours. 24 hours later, 1 mg/mL of MTT reagent was treated, and 2 hours later, formazan generated in cells due to MTT treatment was dissolved in DMSO and absorbance was measured at 570 nm. As a positive control, tacrolimus (1 and 10 μg/mL) was treated.
그 결과, 유산균 발효 타임 추출물과 타임 추출물이 인간각질형성세포의 세포 생존율에 크게 영향을 미치지 않는 것을 확인하였으며, 양성대조군으로 타크롤리무스 대비 다소 인간각질형성세포의 생존을 촉진하는 결과를 나타냈다. 상기 결과를 통해 유산균 발효 타임 추출물이 인체에 독성을 나타내지 않음을 확인할 수 있었다(도 1 참조).As a result, it was confirmed that the lactic acid bacteria fermented thyme extract and the thyme extract did not significantly affect the cell viability of human keratinocytes, and as a positive control, it showed a result of promoting the survival of human keratinocytes somewhat compared to tacrolimus. Through the above results, it was confirmed that the lactic acid bacteria fermented thyme extract was not toxic to the human body (see FIG. 1).
실험예 2. 아토피 인자인 TARC 발현 억제Experimental Example 2. Inhibition of expression of TARC, an atopic factor
인간각질형성세포(HaCaT 5×105/well)를 FBS DMEM 배지에서 24시간 배양 후, FBS 없는 DMEM 배지에 실시예 1에서 제조한 유산균 발효 타임 추출물(10 및 100㎍/㎖)과 TNF-a 및 IFN-r(10ng/mL)을 희석하여 동시 처리하고, 추가 배양하였다. 24시간 경과 후, 각 well의 상층액을 human TARC ELISA kit를 사용하여 제조사 프로토콜에 따라 정량하였다.After culturing human keratinocytes (HaCaT 5×10 5 /well) in FBS DMEM medium for 24 hours, the lactic acid bacteria fermentation thyme extract (10 and 100 μg/ml) and TNF-a prepared in Example 1 in DMEM medium without FBS. and IFN-r (10 ng/mL) diluted and co-treated, and further cultured. After 24 hours, the supernatant of each well was quantified according to the manufacturer's protocol using a human TARC ELISA kit.
그 결과, 도 2에 나타낸 바와 같이, TNF-a 및 IFN-r에 의해 과발현된 TARC 생성량이 본 발명의 유산균 발효 타임 추출물 처리로 유의적으로 억제됨을 확인할 수 있었다. As a result, as shown in FIG. 2 , it was confirmed that the amount of TARC overexpressed by TNF-a and IFN-r was significantly inhibited by the treatment with the lactic acid bacteria fermented thyme extract of the present invention.
구체적으로 양성대조군으로 사용된 타크롤리무스(tacrolimus)(10㎍/mL) 처리군 대비 유산균 발효 타임 추출물 처리군(10㎍/mL)에서 63% 감소된 TARC 생성량이 측정되었으며, 유산균 발효 처리되지 않은 타임추출물 대비 각각 52%(10㎍/mL) 및 49%(100㎍/mL) 감소된 생성량이 측정되어 본 발명의 유산균 발효 타임 추출물이 아토피에 매우 효과적이고, 종래 아토피에 사용되는 면역억제 화합물인 타크롤리무스(tacrolimus) 보다 우수한 효과가 있어 이를 대체할 수 있는 천연 약제로 사용할 수 있음을 확인할 수 있었다. Specifically, a 63% reduction in TARC production was measured in the lactobacillus fermentation thyme extract treated group (10㎍/mL) compared to the tacrolimus (10㎍/mL) treated group used as a positive control. Compared to the thyme extract, the production amount reduced by 52% (10㎍/mL) and 49% (100㎍/mL), respectively, was measured, so that the lactic acid bacteria fermented thyme extract of the present invention is very effective for atopy, and is an immunosuppressive compound used for conventional atopy. It was confirmed that it can be used as a natural drug that can replace it because it has a superior effect than tacrolimus.
실험예 3. 아토피 인자인 MDC 발현 억제Experimental Example 3. Inhibition of MDC expression, atopic factor
인간각질형성세포(HaCaT 5×105/well)를 FBS DMEM 배지에서 24시간 배양 후, FBS 없는 DMEM 배지에 실시예 1에서 제조한 유산균 발효 타임 추출물(10 및 100㎍/㎖)과 TNF-a 및 IFN-r(10ng/mL)을 희석하여 동시 처리하고, 추가 배양하였다. 24시간 경과 후, 각 well의 상층액을 human MDC ELISA kit를 사용하여 제조사 프로토콜에 따라 정량하였다.After culturing human keratinocytes (HaCaT 5×10 5 /well) in FBS DMEM medium for 24 hours, the lactic acid bacteria fermentation thyme extract (10 and 100 μg/ml) and TNF-a prepared in Example 1 in DMEM medium without FBS. and IFN-r (10 ng/mL) diluted and co-treated, and further cultured. After 24 hours, the supernatant of each well was quantified according to the manufacturer's protocol using a human MDC ELISA kit.
그 결과, 도 3에 나타낸 바와 같이, TNF-a 및 IFN-r에 의해 과발현된 MDC 생성량이 본 발명의 유산균 발효 타임 추출물 처리로 유의적으로 억제됨을 확인할 수 있었다. As a result, as shown in FIG. 3, it was confirmed that the amount of MDC overexpressed by TNF-a and IFN-r was significantly inhibited by the treatment with the lactic acid bacteria fermented thyme extract of the present invention.
구체적으로 양성대조군으로 사용된 타크롤리무스(tacrolimus)(10㎍/mL) 처리군 대비 유산균 발효 타임 추출물 처리군(10㎍/mL)에서 67% 감소된 MDC 생성량이 측정되었으며, 유산균 발효 처리되지 않은 타임추출물 대비 각각 27%(10㎍/mL) 및 56%(100㎍/mL) 감소된 MDC 생성량이 측정되어 본 발명의 유산균 발효 타임 추출물이 아토피에 매우 효과적이고, 종래 아토피에 사용되는 면역억제제인 타크롤리무스 화합물에 비해 우수한 효과가 있어 이를 대체할 수 있는 천연 약제로 사용할 수 있음을 확인할 수 있었다. Specifically, a 67% reduction in MDC production was measured in the lactic acid bacteria fermentation thyme extract treated group (10㎍/mL) compared to the tacrolimus (10㎍/mL) treated group used as a positive control, The amount of MDC production reduced by 27% (10㎍/mL) and 56% (100㎍/mL) compared to the thyme extract, respectively, was measured, so that the lactic acid bacteria fermented thyme extract of the present invention is very effective for atopy, and is an immunosuppressant used for conventional atopy. It was confirmed that it can be used as a natural drug that can replace it because it has an excellent effect compared to the tacrolimus compound.
실험예 4. 염증성 사이토카인 생성 억제Experimental Example 4. Inhibition of inflammatory cytokine production
아토피성 피부염은 염증반응을 통해 더욱 확대되며, 피부 발적, 피부 과민반응, 또는 광과민 반응 등을 통해 임상적으로 아토피성 피부염 증상이 더욱 악화되는 경향을 보인다. 따라서, 아토피성 피부염이 발생한 피부 상태의 효과적인 개선을 위해서 염증반응의 조기 차단은 여드름 증상의 치료에 있어 매우 중요한 요인이다. Atopic dermatitis is further expanded through an inflammatory reaction, and symptoms of atopic dermatitis clinically tend to worsen through skin redness, skin hypersensitivity reaction, or photosensitivity reaction. Therefore, in order to effectively improve the skin condition in which atopic dermatitis has occurred, early blocking of the inflammatory reaction is a very important factor in the treatment of acne symptoms.
유산균 발효 타임 추출물이 염증반응과 면역 활성을 억제하는 효과를 아래와 같은 방법으로 검증하였다. The effect of lactic acid bacteria fermented thyme extract in suppressing inflammatory response and immune activity was verified in the following way.
아토피성 피부염에 의해 발생되는 염증반응 메커니즘에서 IL-6 등의 염증매개 물질들이 관여하게 되며, 염증매개 물질들의 작용에 의해 염증반응이 증폭되게 되며, IL-6의 발현을 억제하는 경우 염증반응을 효과적으로 차단할 수 있게 된다.In the inflammatory reaction mechanism caused by atopic dermatitis, inflammatory mediators such as IL-6 are involved, and the inflammatory response is amplified by the action of inflammatory mediators. can be effectively blocked.
아토피성 피부염이 IL-6 단백질의 발현에 미치는 영향을 검증하기 위해 인간각질형성세포(HaCaT)를 대상을 실험을 수행하였다. HaCaT 세포를 96 well plate에 5.0×105 농도로 접종한 후, UVB를 144mJ/cm2 조건으로 조사하고, FBS 없는 DMEM에 유산균 발효 타임 추출물(각각 1, 10, 및 100㎍/㎖)을 처리하고, 24 시간동안 추가 배양하였다. 배양이 끝나고, 상등액을 취하여 IL-6 단백질 생성량을 효소결합 면역흡수 분석법(ELISA, Enzyme-Linked Immunosorbent Assay) 키트를 사용하여 분석하였다.In order to verify the effect of atopic dermatitis on the expression of IL-6 protein, an experiment was performed on human keratinocytes (HaCaT). HaCaT cells were inoculated in a 96 well plate at a concentration of 5.0 × 10 5 , UVB was irradiated at 144 mJ/cm 2 conditions, and lactic acid bacteria fermentation thyme extract (1, 10, and 100 μg/ml, respectively) was treated in DMEM without FBS. and further incubated for 24 hours. After incubation, the supernatant was taken and the amount of IL-6 protein produced was analyzed using an enzyme-linked immunosorbent assay (ELISA, Enzyme-Linked Immunosorbent Assay) kit.
도 4를 참조하면, 유산균 발효 타임 추출물 처리시 UVB로 유도된 IL-6 단백질의 생성이 억제되는 것으로 나타났으며, 이러한 IL-6 단백질 생성 억제능은 농도 의존적인 것으로 나타났다.Referring to FIG. 4 , it was found that UVB-induced IL-6 protein production was inhibited during treatment with lactic acid bacteria fermented thyme extract, and this ability to inhibit IL-6 protein production was found to be concentration-dependent.
구체적으로 유산균 발효 처리되지 않은 타임추출물은 자외선 조사군 대비 각각 1, 10, 및 100㎍/㎖ 용량에서 44%, 61%, 및 77% 감소된 IL-6 생성량이 측정되었고, 본 발명의 유산균 발효 타임 추출물은 자외선 조사군 대비 각각 1, 10, 및 100㎍/㎖ 용량에서 51%, 70%, 및 84%로 더욱 감소된감소된 IL-6 생성량이 측정되었다. 상기와 같은 실험결과를 통해 본 발명의 유산균 발효 타임 추출물이 아토피성 면역 또는 염증 반응의 억제에 매우 효과적이고, 유산균 발효를 통해 타임 추출물이 종래 갖고 있는 면역 또는 염증 반응을 획기적으로 개선하여 사용할 수 있음을 확인할 수 있었다.Specifically, in the thyme extract not subjected to lactic acid fermentation, 44%, 61%, and 77% reduced IL-6 production was measured at 1, 10, and 100 μg/ml doses, respectively, compared to the UV irradiation group, and the lactic acid bacteria fermentation of the present invention The thyme extract was measured for reduced IL-6 production, which was further reduced to 51%, 70%, and 84% at doses of 1, 10, and 100 μg/ml, respectively, compared to the UV-irradiated group. Through the above experimental results, the lactic acid bacteria fermented thyme extract of the present invention is very effective in suppressing atopic immunity or inflammatory response, and through lactic acid bacterium fermentation, the thyme extract can dramatically improve and use the conventional immune or inflammatory response. was able to confirm
실험예 5. MMP-1 생성 억제Experimental Example 5. Inhibition of MMP-1 production
40mm 세포 배양 접시에 2㎖의 DMEM 배양액을 넣고, 인간섬유아세포를 약 1.2x105의 농도로 접종한 후, 37℃, 5% CO2 환경에서 24시간 동안 배양하였다. 그 후, UVB를 144mJ/cm2 조건으로 조사한 후, 실시예 1에서 제조한 유산균 발효 타임 추출물(각각 1, 10, 및 100㎍/㎖)을 포함한 배지로 교체하여 3일간 배양하였다. 이후 배양액을 회수(harvest)하여 4℃, 7,500rpm 환경에서 5분간 원심분리(centrifuge)하여 ELISA 방법을 이용하여 MMP-1(Human Total MMP-1 kit, R&D Systems, Inc., Minneapolis, MN, USA) 단백질의 발현량 변화를 확인하였다.2ml of DMEM culture solution was placed in a 40mm cell culture dish, and human fibroblasts were inoculated at a concentration of about 1.2x10 5 , and then cultured at 37° C., 5% CO 2 environment for 24 hours. Thereafter, after irradiating UVB with 144mJ/cm 2 conditions, the medium containing the lactic acid bacteria fermentation thyme extract prepared in Example 1 (1, 10, and 100 μg/ml, respectively) was replaced and cultured for 3 days. After that, the culture medium was harvested and centrifuged for 5 minutes at 4° C., 7,500 rpm, and MMP-1 (Human Total MMP-1 kit, R&D Systems, Inc., Minneapolis, MN, USA) using an ELISA method. ) to confirm the change in the expression level of the protein.
도 5를 참조하면, 144mJ/cm2 용량으로 자외선 조사 처리한 인간섬유아세포는 자외선 무처리 군에 비해 MMP-1 단백질의 생성이 3배 정도 증가하였다. 이렇게 자외선 조사 처리로 인한 MMP-1 단백질의 생성 증가는 타임 추출물의 처리에 의해서 감소하는 현상을 나타내었는데, 자외선 조사군 대비 각각 1, 10, 및 100㎍/㎖ 용량에서 2%, 14%, 및 25% 감소된 MMP-1 단백질의 생성 저해 효과를 나타내었다. Referring to FIG. 5 , human fibroblasts treated with UV irradiation at a dose of 144mJ/cm 2 increased the production of MMP-1 protein by about 3 times compared to the UV-untreated group. Thus, the increase in the production of MMP-1 protein due to the UV irradiation treatment showed a phenomenon that decreased by the treatment of the thyme extract, 2%, 14%, and It exhibited an inhibitory effect on the production of MMP-1 protein reduced by 25%.
한편, 상기와 같은 타임의 MMP-1 단백질의 생성 억제 효과는 유산균 발효 처리에 의해 더욱 증가되는 양상을 보였는데, 본 발명의 유산균 발효 타임 추출물 처리군에서는 자외선 조사군 대비 각각 1, 10, 및 100㎍/㎖ 용량에서 9%, 37%, 및 54%로 더욱 감소된 MMP-1 단백질의 생성 저해 효과를 나타내었다.On the other hand, the effect of inhibiting the production of MMP-1 protein of thyme as described above was further increased by the lactic acid bacterium fermentation treatment. In the μg/ml dose, the inhibitory effect on the production of MMP-1 protein was further reduced to 9%, 37%, and 54%.
상기와 같은 결과를 통해, 본 발명의 유산균 발효 타임 추출물은 주름생성의 원인이 되는 MMP-1 단백질의 생성을 효과적으로 억제하여 피부 주름의 예방 및 개선에 효과적으로 사용될 수 있음을 알 수 있었다. Through the above results, it was found that the lactic acid bacteria fermented thyme extract of the present invention can be effectively used to prevent and improve skin wrinkles by effectively inhibiting the production of MMP-1 protein, which is the cause of wrinkles.
실험예 6. 타입 1 프로콜라겐 단백질의 합성 촉진Experimental Example 6. Promotion of synthesis of
40mm 세포 배양 접시에 2㎖의 DMEM 배양액을 넣고, 인간섬유아세포를 약 1.2x105의 농도로 접종한 후, 37℃, 5% CO2 환경에서 24시간 동안 배양하였다. 그 후, UVB를 144mJ/cm2 조건으로 조사한 후, 실시예 1에서 제조한 유산균 발효 타임 추출물(각각 10, 및 100㎍/㎖)을 포함한 배지로 교체하여 배양하였다. 이후 배양액을 회수(harvest)하여 4℃, 7500rpm 환경에서 5분간 원심분리(centrifuge)하고 ELISA 방법을 이용하여 타입 1 프로콜라겐(Procollagen Type I C Peptide EIA Kit, Takara, Shiga, Japan)의 단백질 발현량 변화를 확인하였다.2ml of DMEM culture solution was placed in a 40mm cell culture dish, and human fibroblasts were inoculated at a concentration of about 1.2x10 5 , and then cultured at 37° C., 5% CO 2 environment for 24 hours. Thereafter, after irradiating UVB with 144mJ/cm 2 conditions, it was cultured by replacing the medium containing the lactic acid bacteria fermentation thyme extract prepared in Example 1 (10, and 100 μg/ml, respectively). Thereafter, the culture medium was harvested, centrifuged for 5 minutes at 4°C and 7500 rpm, and the protein expression level of
도 6을 참조하면, 144mJ/cm2 용량으로 자외선 조사 처리한 인간섬유아세포는 자외선 무처리군에 비해 타입 1 프로콜라겐 단백질의 생성이 약 2.3배 정도 감소하였다. 이렇게 자외선 조사 처리로 인한 타입 1 프로콜라겐 단백질의 생성 감소는 타임 추출물의 처리에 의해서 다시 증가하는 현상을 나타내었다. 이 때, 타임 추출물 처리군은 무처리군과 비교할 때 발현량 증가 효과를 나타내는데, 각각 10, 및 100㎍/㎖ 용량에서 109% 및 138% 개선된 타입 1 프로콜라겐 단백질의 생성 증가 효과를 나타내었다.Referring to FIG. 6 , human fibroblasts treated with UV irradiation at a dose of 144mJ/cm 2 reduced the production of
한편, 상기와 같은 타임의 타입 1 프로콜라겐 단백질의 생성 증가 효과는 유산균 발효 처리에 의해 더욱 증가되는 양상을 보였는데, 본 발명의 유산균 발효 타임 추출물 처리군에서는 각각 10, 및 100㎍/㎖ 용량에서 무처리군 대비 153% 및 234%로 더욱 증가된 타입 1 프로콜라겐 단백질의 생성 증가 효과를 나타내었다.On the other hand, the effect of increasing the production of
상기와 같은 결과를 통해, 본 발명의 유산균 발효 타임 추출물은 주름생성을 억제하는 타입 1 프로콜라겐 단백질의 생성을 효과적으로 증진하여 피부 주름의 예방 및 개선에 효과적으로 사용될 수 있음을 알 수 있었다. Through the above results, it was found that the lactic acid bacteria fermented thyme extract of the present invention can be effectively used to prevent and improve skin wrinkles by effectively promoting the production of
상기와 같은 결과를 통해서 본 발명의 유산균 발효 타임 추출물을 유효성분으로 함유하는 조성물은 아토피 인자인 TARC 및 MDC 단백질의 생성을 효과적으로 억제하고, IL-6와 같은 염증성 사이토카인 생성을 억제하여 아토피성 면역 또는 염증 반응의 억제에 매우 효과적이며, MMP-1 단백질 생성 억제 및 타입 1 프로콜라겐 단백질 생성 증진을 통해 피부 주름의 예방 및 개선에도 효과적임을 확인할 수 있음을 알 수 있었다. Through the above results, the composition containing the lactic acid bacteria fermented thyme extract of the present invention as an active ingredient effectively inhibits the production of TARC and MDC proteins, which are atopic factors, and inhibits the production of inflammatory cytokines such as IL-6 to prevent atopic immunity. Alternatively, it was found that it can be confirmed that it is very effective in inhibiting the inflammatory response, and is also effective in preventing and improving skin wrinkles through inhibition of MMP-1 protein production and promotion of
이상으로 본 발명의 특정한 부분을 상세히 기술하였는 바, 당업계의 통상의 지식을 가진 자에게 있어서 이러한 구체적인 기술은 단지 바람직한 구현예일 뿐이며, 이에 본 발명의 범위가 제한되는 것이 아닌 점은 명백하다. 따라서, 본 발명의 실질적인 범위는 첨부된 청구항과 그의 등가물에 의하여 정의된다고 할 것이다.As described above in detail a specific part of the present invention, for those of ordinary skill in the art, this specific description is only a preferred embodiment, and it is clear that the scope of the present invention is not limited thereto. Accordingly, the substantial scope of the present invention will be defined by the appended claims and their equivalents.
Claims (7)
상기 조성물은 TARC(Thymus and activation-regulated chemokine) 및 MDC(macrophage-derived chemokine) 생성을 억제하는 것을 특징으로 하는,
아토피 피부염 및 피부주름 개선용 화장료 조성물.Contains Lactobacillus plantarum (Accession No.: KCTC 14013BP) fermented thyme extract as an active ingredient,
The composition is characterized in that it inhibits TARC (Thymus and activation-regulated chemokine) and MDC (macrophage-derived chemokine) production,
A cosmetic composition for improving atopic dermatitis and skin wrinkles.
상기 조성물은 MMP-1 단백질 생성을 억제하고, 타입 1 프로콜라겐 단백질 생성을 증가시키는 것을 특징으로 하는 아토피 피부염 및 피부주름 개선용 화장료 조성물.The method of claim 1,
The composition inhibits MMP-1 protein production and a cosmetic composition for improving atopic dermatitis and skin wrinkles, characterized in that it increases the production of type 1 procollagen protein.
상기 조성물은 TARC(Thymus and activation-regulated chemokine) 및 MDC(macrophage-derived chemokine) 생성을 억제하는 것을 특징으로 하는,
아토피 피부염 및 피부주름 개선용 식품 조성물.Contains Lactobacillus plantarum (Accession No.: KCTC 14013BP) fermented thyme extract as an active ingredient,
The composition is characterized in that it inhibits TARC (Thymus and activation-regulated chemokine) and MDC (macrophage-derived chemokine) production,
A food composition for improving atopic dermatitis and skin wrinkles.
상기 조성물은 TARC(Thymus and activation-regulated chemokine) 및 MDC(macrophage-derived chemokine) 생성을 억제하는 것을 특징으로 하는,
아토피 피부염 치료 또는 예방용 약학적 조성물.Contains Lactobacillus plantarum (Accession No.: KCTC 14013BP) fermented thyme extract as an active ingredient,
The composition is characterized in that it inhibits TARC (Thymus and activation-regulated chemokine) and MDC (macrophage-derived chemokine) production,
A pharmaceutical composition for treating or preventing atopic dermatitis.
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| SEO, SEULA 외 6명. Thymus vulgaris Suppresses Atopic Dermatitis-Like Symptoms in Human Keratinocyte. KMB 2019 46th Annual Meeting & International Symposium, 2019.06.23* |
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