KR102501800B1 - A composition with anti-inflammation activity and skin elasticity improvement efficacy comprising mixed ferment extracts of natural products - Google Patents
A composition with anti-inflammation activity and skin elasticity improvement efficacy comprising mixed ferment extracts of natural products Download PDFInfo
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- KR102501800B1 KR102501800B1 KR1020210000628A KR20210000628A KR102501800B1 KR 102501800 B1 KR102501800 B1 KR 102501800B1 KR 1020210000628 A KR1020210000628 A KR 1020210000628A KR 20210000628 A KR20210000628 A KR 20210000628A KR 102501800 B1 KR102501800 B1 KR 102501800B1
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- inflammatory
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Abstract
천연물 복합 발효 추출물을 포함하는 항염증, 피부탄력 증진 효능을 갖는 조성물에 관한 것이다. 일 양상에 따른 조성물은 녹차, 생강 및 편백 혼합 추출물을 발효시킨 복합 발효 추출물을 유효성분으로 포함하며, 염증 관련 단백질의 발현 억제를 통한 항염증 효과가 우수하고, 피부탄력을 유지시키는 단백질인 엘라스틴을 분해하는 엘라스타제 활성을 효과적으로 억제하여 피부탄력 증진 및 피부주름 개선 효과를 제공할 수 있다. It relates to a composition having anti-inflammatory and skin elasticity enhancing effects including a natural product complex fermented extract. The composition according to one aspect includes, as an active ingredient, a complex fermented extract obtained by fermenting a mixed extract of green tea, ginger, and cypress, and has an excellent anti-inflammatory effect by suppressing the expression of inflammation-related proteins and elastin, a protein that maintains skin elasticity. By effectively suppressing the decomposing elastase activity, it is possible to provide skin elasticity enhancement and skin wrinkle improvement effects.
Description
천연물 복합 발효 추출물을 포함하는 항염증, 피부탄력 증진 효능을 갖는 조성물에 관한 것이다.It relates to a composition having anti-inflammatory and skin elasticity enhancing effects including a natural product complex fermented extract.
최근 생활습관의 변화, 화학물질에 대한 빈번한 노출, 환경오염의 증가 및 현대인의 정신적 스트레스 등으로 인하여 아토피 피부염과 같은 피부 염증 질환의 발병이 늘어나고 있다. 피부는 물리적 또는 화학적 자극으로부터 신체를 보호하는 물리적 장벽으로서 항상성을 유지하고, 가장 표면층에 해당하는 표피는 외부 환경적 요인에 따라 사이토카인 및 염증성 인자를 생산하여 염증반응과 면역반응을 조절한다. 이러한 피부 염증 질환의 치료에는 면역계 염증반응 조절을 위해 스테로이드 제제가 주로 사용되지만, 사용에 따라 내성이 생기고 피부장벽을 약화시켜 장기간 치료가 어렵고, 혈관을 확장시켜 이차 감염 등의 심각한 부작용이 있다. 이에, 부작용이나 역효과가 없는 천연물을 이용한, 안전하고 효과적인 피부 상태 개선용 제제에 대한 요구가 어느 때보다도 증가하고 있다.Recently, the occurrence of skin inflammatory diseases such as atopic dermatitis is increasing due to changes in lifestyle, frequent exposure to chemicals, increased environmental pollution, and mental stress of modern people. The skin maintains homeostasis as a physical barrier that protects the body from physical or chemical stimuli, and the epidermis, which is the most superficial layer, produces cytokines and inflammatory factors according to external environmental factors to regulate inflammatory and immune responses. In the treatment of inflammatory skin diseases, steroid preparations are mainly used to control the inflammatory response of the immune system, but resistance develops with use and weakens the skin barrier, making long-term treatment difficult and dilating blood vessels, resulting in serious side effects such as secondary infection. Accordingly, the demand for safe and effective preparations for improving skin conditions using natural products without side effects or adverse effects is increasing more than ever.
한편, 천연물로부터 유용생리활성 성분을 추출하기 위해서는 유산균 등의 유익 미생물들의 대사작용을 이용한 천연물 발효 공정이 효과적일 수 있다. 발효(fermentation)란, 학문적인 원래의 뜻은 미생물이 탄수화물을 에너지원으로 할 때 그 탄소결합을 끊어서 알코올, 유기산, 탄산가스 등이 생성되는 현상을 말한다. 유산균은 탄수화물 발효를 통해 최종 대사산물로 유산을 생산하는 균으로, 유산균 발효를 통한 천연 추출물 함유 활성 성분 및 생리활성의 증가를 목적으로 다양한 연구가 활발하게 진행되고 있다.On the other hand, in order to extract useful physiologically active components from natural products, a natural product fermentation process using the metabolism of beneficial microorganisms such as lactic acid bacteria may be effective. Fermentation, in its original scientific meaning, refers to a phenomenon in which alcohol, organic acid, carbon dioxide, etc. are produced by breaking the carbon bond when microorganisms use carbohydrates as an energy source. Lactic acid bacteria are bacteria that produce lactic acid as a final metabolite through carbohydrate fermentation, and various studies are being actively conducted for the purpose of increasing active ingredients and physiological activities containing natural extracts through lactic acid bacteria fermentation.
본 발명자들은 천연물인 녹차, 생강 및 편백 복합 추출물을 유산균을 이용하여 발효시키고, 상기 천연물 복합 발효 추출물은 비발효 추출물에 비해 항염 효과가 현저히 높을 뿐만 아니라, 피부탄력을 증진시켜 항염증 용도, 피부탄력 증진 용도 또는 피부주름 개선 용도로 사용할 수 있음을 확인함으로써 본 발명을 완성하였다.The present inventors fermented natural product complex extracts of green tea, ginger, and cypress using lactic acid bacteria, and the natural product complex fermented extract not only has a significantly higher anti-inflammatory effect than non-fermented extracts, but also enhances skin elasticity for anti-inflammatory use and skin elasticity. The present invention was completed by confirming that it can be used for enhancement or skin wrinkle improvement.
일 양상은 녹차, 생강 및 편백 혼합 추출물을 발효시킨 복합 발효 추출물을 포함하는 피부 상태 개선용 화장료 조성물을 제공하는 것이다.One aspect is to provide a cosmetic composition for improving skin condition comprising a complex fermented extract obtained by fermenting a mixed extract of green tea, ginger and cypress.
다른 양상은 녹차, 생강 및 편백 혼합 추출물을 발효시킨 복합 발효 추출물을 포함하는 피부 상태 개선용 식품 조성물을 제공하는 것이다.Another aspect is to provide a food composition for improving skin condition comprising a complex fermented extract obtained by fermenting a mixed extract of green tea, ginger and cypress.
또 다른 양상은 녹차, 생강 및 편백 혼합 추출물을 발효시킨 복합 발효 추출물을 포함하는 항염증용 약학적 조성물을 제공하는 것이다.Another aspect is to provide an anti-inflammatory pharmaceutical composition comprising a complex fermented extract obtained by fermenting a mixed extract of green tea, ginger and cypress.
이하, 본 발명에 대해 상세히 설명한다.Hereinafter, the present invention will be described in detail.
일 양상은 녹차, 생강 및 편백 혼합 추출물을 발효시킨 복합 발효 추출물을 포함하는 피부 상태 개선용 화장료 조성물을 제공한다.One aspect provides a cosmetic composition for improving skin condition comprising a complex fermented extract obtained by fermenting a mixed extract of green tea, ginger and cypress.
상기 피부 상태 개선은 항염증, 피부탄력 증진 및 피부주름 개선으로 이루어진 군으로부터 선택되는 하나 이상일 수 있다.The skin condition improvement may be at least one selected from the group consisting of anti-inflammatory, skin elasticity enhancement, and skin wrinkle improvement.
상기 녹차, 생강 및 편백 혼합 추출물을 발효시킨 복합 발효 추출물은 높은 NO 생성 저해능을 보이고, 세포의 염증을 촉진시키는 사이토카인인 TNF-α 및 IL-6의 발현을 감소시켜 항염증 효과가 우수하다. 뿐만 아니라, 엘라스타제(elastase) 활성을 억제하여 피부탄력 및 주름을 효과적으로 예방 또는 개선할 수 있다.The complex fermented extract obtained by fermenting the green tea, ginger, and cypress mixed extract exhibits high NO production inhibition and has excellent anti-inflammatory effect by reducing the expression of TNF-α and IL-6, which are cytokines that promote cell inflammation. In addition, it is possible to effectively prevent or improve skin elasticity and wrinkles by inhibiting elastase activity.
따라서, 상기 복합 발효 추출물을 포함하는 조성물은 항염증, 피부탄력 증진 또는 피부주름 개선용 조성물로 사용될 수 있다.Therefore, the composition containing the complex fermented extract can be used as a composition for anti-inflammatory, skin elasticity enhancement or skin wrinkle improvement.
용어 "항염증"은 염증을 억제하는 작용을 의미하는 것으로, 염증반응의 조절은 대단히 복잡한 것으로 알려져 있는데, 이는 생체 내 복구체계의 증강 및 손상을 감소시키기 위한 것으로 알려져 있다. 그러나 반복되는 조직의 손상이나 재생에 의해 염증반응이 지속되면, 염증관련 세포에서 ROS와 RNS가 과다 생성되고 그 결과로 영구적인 유전자의 변형이 야기된다. 이처럼 ROS와 RNS는 생체 내 여러 가지 세포의 작용을 조절하는 염증 반응과 깊이 관련되어 있다. 주요한 염증성 사이토카인으로는 TNF-α(tumor necrosis factor-α), IL-1(interleukin-1), IL-6 및 IL-8 등이 있다. The term "anti-inflammatory" refers to an action of suppressing inflammation, and the regulation of the inflammatory response is known to be very complex, which is known to enhance and reduce damage to the repair system in vivo. However, if the inflammatory response continues due to repeated tissue damage or regeneration, ROS and RNS are excessively produced in inflammation-related cells, resulting in permanent genetic modification. As such, ROS and RNS are deeply related to the inflammatory response that regulates the actions of various cells in vivo. Major inflammatory cytokines include tumor necrosis factor-α (TNF-α), interleukin-1 (IL-1), IL-6, and IL-8.
일 구체예에서, 일 양상에 따른 조성물은 NO 생성을 억제하고, 염증관련인자인 TNF-α 및 IL-6의 발현을 현저히 억제하여 높은 항염증 활성을 가짐을 확인하였다.In one embodiment, the composition according to one aspect was confirmed to have high anti-inflammatory activity by inhibiting NO production and significantly inhibiting the expression of TNF-α and IL-6, which are inflammation-related factors.
용어 "피부탄력"은 진피층에 존재하는 엘라스틴(elastin)으로 구성된 탄력 섬유에 의해 나타나는 탄성을 말하며, 이러한 탄력 섬유는 고무와 같이 매우 낮은 탄성 계수를 가지고 있어서 작은 힘에 의해서도 쉽게 변형되고, 그 힘이 제거되었을 때는 쉽게 원형으로 되돌아 온다. 또한, 탄력 섬유는 엘라스틴이라는 무정형의 기질에 미원섬유(microfibrils)들이 박혀 있는 형태를 띠고 있으며, 엘라스틴은 라이신에서 유래한 데스모신(desmosine)과 아이소데스모신(isodesmosine)이라는 탄력 섬유에서만 발견되는 아주 독특한 아미노산으로 구성된 단백질이다. 이러한 데스모신과 아이소데스모신 등은 긴 펩타이드 사슬 안에서 가교(cross-links)를 형성하고 있는데, 이러한 구조가 엘라스틴으로 하여금 고무와 같은 성질을 갖게 한다.The term "skin elasticity" refers to the elasticity exhibited by elastic fibers composed of elastin present in the dermal layer, and these elastic fibers have a very low modulus of elasticity like rubber, so they are easily deformed even by a small force, and the force is When removed, it easily returns to its original form. In addition, elastic fibers have a form in which microfibrils are embedded in an amorphous matrix called elastin. It is a protein composed of amino acids. Such desmosin and isodesmosin form cross-links in a long peptide chain, and this structure gives elastin a rubber-like property.
용어 "피부탄력 개선"이란 엘라스틴으로 구성된 탄력 섬유가 콜라겐이라고 하는 교원섬유와 함께 존재하는데, 엘라스틴과 콜라겐이 충분히 존재하는 상태에서 피부 탄력이 유지되는 것을 의미한다.The term "improvement of skin elasticity" means that elastic fibers composed of elastin exist together with collagen fibers called collagen, and skin elasticity is maintained in a state in which elastin and collagen are sufficiently present.
용어 "피부주름"이란 피부가 쇠하여 생긴 잔줄을 의미하는데, 유전자에 의한 원인, 피부 진피에 존재하는 콜라겐과 엘라스틴의 감소, 외부환경 등에 의해 유발될 수 있다.The term "skin wrinkles" refers to fine lines caused by the decline of the skin, which may be caused by genes, reduction of collagen and elastin present in the skin dermis, external environment, and the like.
용어 "피부주름 개선"이란 피부에 주름이 생성되는 것을 억제 또는 저해하거나, 이미 생성된 주름을 완화시키는 것을 의미한다.The term "improvement of skin wrinkles" means suppressing or inhibiting the formation of wrinkles on the skin, or alleviating wrinkles already formed.
녹차용 차나무(학명 Camellia sinensis)는 상록의 활엽관목으로서, 폴리페놀류, 다당류, 비타민 등 다양한 영양 성분들을 함유하고 있을 뿐 아니라 여러 질병의 위험을 경감시키는 효과가 있다고 알려져 있다. 녹차 중에 존재하는 폴리페놀은 녹차의 주요 활성성분으로 주목받고 있으며, 특히 이 중 카테킨류는 체내 활성산소를 제거하는 항산화 효과가 비타민 E의 50배, 비타민 C의 100배나 높은 것으로 알려져 있다.Tea tree for green tea (scientific name Camellia sinensis ) is an evergreen broad-leaved shrub, and is known to contain various nutrients such as polyphenols, polysaccharides, and vitamins, as well as to reduce the risk of various diseases. Polyphenols present in green tea are attracting attention as the main active ingredient of green tea, and among them, catechins are known to have 50 times higher antioxidant effects than vitamin E and 100 times higher than vitamin C.
상기 녹차는 녹차잎, 줄기 및 씨로 이루어진 군으로부터 선택되는 하나 이상, 바람직하게는 녹차잎을 이용하여 추출물을 제조할 수 있으나, 이에 제한되는 것은 아니다. 예를 들어, 녹차잎을 세척 및 건조시킨 것 또는 건조된 녹차잎을 잘게 분쇄하여 분말 형태로 만든 것을 추출에 사용할 수 있다. 상기 녹차잎은 녹차 유효성분의 함량을 증가시킬 수 있도록 100 내지 300℃로 가열하여 덖은 후 건조하고, 잘게 분쇄하여 분말 형태로 만든 것일 수 있다.The green tea extract may be prepared using at least one selected from the group consisting of green tea leaves, stems and seeds, preferably using green tea leaves, but is not limited thereto. For example, green tea leaves washed and dried or dried green tea leaves pulverized into a powder form may be used for extraction. The green tea leaves may be roasted by heating at 100 to 300° C. to increase the content of green tea active ingredients, dried, and finely pulverized to form a powder.
생강(학명 Zingiber officinale)은 생강과에 속하는 아열대 및 열대성 다년생 식물로서, 우리나라뿐만 아니라 세계적으로 널리 이용되고 있는 향신료의 하나이다. 생강은 인체에 유익한 여러 가지 성분이 함유되어 있어 차(tea)나 약재로 널리 사용되어오고 있다. 한방에서는 생강의 뿌리줄기 말린 것을 건강(乾薑)이라는 약재로 사용하고 있는데, 소화불량, 구토설사 등에 효과가 있고, 혈액 순환을 촉진하며, 항염증과 진통 효과를 지니고 있다고 알려져 있다.Ginger (scientific name Zingiber officinale ) is a subtropical and tropical perennial plant belonging to the ginger family, and is one of the spices widely used not only in Korea but also worldwide. Ginger contains various ingredients beneficial to the human body, so it has been widely used as tea or medicine. In oriental medicine, the dried rhizome of ginger is used as a medicine called health, and it is effective for indigestion, vomiting and diarrhea, promotes blood circulation, and is known to have anti-inflammatory and analgesic effects.
상기 생강은 세척하여 절편의 형태가 되도록 소정 크기로 절단한 것을 추출에 사용할 수 있으나, 이에 제한되는 것은 아니다. 추출에는 절단된 원물뿐만 아니라, 상기 절단한 생강을 압착하여 생강 원액으로 추출한 것을 사용할 수 있다. 생강 원액을 사용하는 경우, 절단된 생강의 압착은 공지의 착즙기를 사용하여 수행될 수 있다.The ginger may be washed and cut to a predetermined size to be in the form of slices, but may be used for extraction, but is not limited thereto. For extraction, not only the cut raw material, but also the cut ginger extracted by compressing the ginger juice may be used. In the case of using ginger undiluted solution, pressing of the cut ginger may be performed using a known juicer.
편백(학명 Chamaecyparis obtusa)은 상록 침엽 교목에 속하고 내한성과 내염성에 약하지만 내공해성에 강한 측백나무과에 속하는 식물이다. 편백은 피톤치드라는 천연 항균물질을 다량 함유하여 알레르기를 포함하는 피부 질환 개선, 항균, 항스트레스, 탈취와 정화 기능 및 면역 기능을 가지는 것으로 알려져 있다.Cypress (scientific name: Chamaecyparis obtusa ) belongs to evergreen needle-leaved trees and is weak in cold resistance and salt resistance, but is strong in pollution resistance. Cypress contains a large amount of natural antibacterial substance called phytoncide, and is known to have skin disease improvement including allergy, antibacterial, antistress, deodorizing and purifying functions, and immune functions.
상기 편백은 편백의 잎, 줄기 및 가지로 이루어진 군으로부터 선택되는 하나 이상, 바람직하게는 편백잎을 이용하여 추출물을 제조할 수 있으나, 이에 제한되는 것은 아니다. 예를 들어, 편백잎을 세척 및 건조시킨 것 또는 건조된 편백잎을 잘게 분쇄하여 분말 형태로 만든 것을 추출에 사용할 수 있다.The cypress may prepare an extract using at least one selected from the group consisting of leaves, stems and branches of cypress, preferably cypress leaves, but is not limited thereto. For example, those made by washing and drying cypress leaves or finely pulverizing dried cypress leaves can be used for extraction.
상기 녹차, 생강 및 편백 혼합 추출물을 발효시킨 복합 발효 추출물은 녹차, 생강 및 편백 원물을 특정 비율로 혼합하여 혼합 추출물을 제조한 이후, 상기 혼합 추출물을 발효시켜 제조하는 것일 수 있다.The complex fermented extract obtained by fermenting the mixed extract of green tea, ginger, and cypress may be prepared by preparing a mixed extract by mixing raw green tea, ginger, and raw cypress in a specific ratio, and then fermenting the mixed extract.
용어 "추출물"은 천연물로부터 분리된 활성성분 즉, 목적하는 활성을 보이는 물질을 의미하며, 물, 유기용매 또는 이들의 혼합용매를 이용하는 추출과정으로 제조될 수 있고, 물, 유기용매 또는 이들의 혼합용매의 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 제형을 포함한다. 또한, 상기 추출물은 상기 추출액을 분획한 것도 포함된다.The term "extract" means an active ingredient separated from a natural product, that is, a substance showing a desired activity, and can be prepared by an extraction process using water, an organic solvent, or a mixture thereof, and water, an organic solvent, or a mixture thereof. It includes all formulations that can be formed using the extract itself and the extract, such as a solvent extract, a dilution or concentrate of the extract, a dried product obtained by drying the extract, a crude or purified product of the extract, or a mixture thereof. In addition, the extract includes a fractionated extract of the extract.
상기 추출은 열수 추출, 침지 추출, 초임계 추출, 아임계 추출, 환류냉각 추출, 수증기 증류, 고압효소분해, 초음파 추출, 용출, 압착 등의 방법을 사용할 수 있으며, 바람직하게는 초음파 추출 방법을 사용할 수 있으나, 이에 제한되는 것은 아니다.The extraction may be carried out by hot water extraction, immersion extraction, supercritical extraction, subcritical extraction, reflux cooling extraction, steam distillation, high-pressure enzyme decomposition, ultrasonic extraction, elution, compression, etc., preferably ultrasonic extraction. It may be, but is not limited thereto.
상기 초음파 추출은 녹차, 생강 및 편백 원물을 각 원물의 1-15 중량배의 추출용매(예를 들어, 물, 유기용매 또는 이들의 혼합용매)와 혼합하여 15 내지 25℃에서 30분 내지 3시간 동안 20 내지 30 kHz의 주파수로 저온 초음파 처리하여 수행할 수 있다. 바람직하게는, 상기 저온 초음파 처리는 17 내지 22℃에서 30분 내지 2시간 동안 22 내지 38 kHz의 주파수로 처리하여 수행할 수 있고, 보다 바람직하게는 20℃에서 1시간 동안 25 내지 27 kHz의 주파수로 처리하여 수행할 수 있다. 상기 초음파 추출은 초음파 에너지 변화의 세기를 조절하여 추출시간은 단축하면서도 추출수율을 높일 수 있으며, 원물 변성 및 파괴 가능성을 최소화할 수 있다.The ultrasonic extraction is performed by mixing green tea, ginger, and cypress raw materials with an extraction solvent (eg, water, organic solvent, or a mixed solvent thereof) of 1-15 times the weight of each raw material, for 30 minutes to 3 hours at 15 to 25 ° C. It can be carried out by low-temperature sonication at a frequency of 20 to 30 kHz during the process. Preferably, the low-temperature ultrasonic treatment can be performed by treatment at a frequency of 22 to 38 kHz for 30 minutes to 2 hours at 17 to 22 ° C., more preferably at a frequency of 25 to 27 kHz for 1 hour at 20 ° C. It can be done by processing with The ultrasonic extraction can increase the extraction yield while shortening the extraction time by adjusting the intensity of the ultrasonic energy change, and can minimize the possibility of denaturation and destruction of the original material.
상기 초음파 추출은 바람직하게는 저온 초음파 처리를 1 내지 5회, 바람직하게는 2 내지 4회, 보다 바람직하게는 3회 반복하여 수행하는 것일 수 있다. Preferably, the ultrasonic extraction may be performed by repeating low-temperature
용어 "발효"는 미생물이 탄수화물을 에너지원으로 할 때 탄수화물의 탄소결합을 끊어서 알코올, 유기산, 탄산가스 등이 생성되는 현상을 의미한다.The term "fermentation" refers to a phenomenon in which alcohol, organic acids, carbon dioxide, etc. are produced by breaking carbon bonds of carbohydrates when microorganisms use carbohydrates as an energy source.
용어 "유산균"은 장에 도달해 젖산을 생성하고 유익균이 증가할 수 있도록 산성으로 변화시켜 주는 살아있는 균을 의미한다. 상기 유산균은, 예를 들어 락토바실러스 플랜타룸(Lactobacillus plantarum), 엔테로코커스 페시움(Enterococcus faecium), 락토바실러스 펜토서스(Lactobacillus pentosus), 락토바실러스 에시도필러스(Lactobacillus acidophilus), 락토바실러스 루테리(Lactobacillus reuteri), 락토바실러스 부크네리(Lactobacillus buchneri) 및 바실러스 코아귤란스(Bacillus coagulans)로 이루어진 군으로부터 선택된 하나 이상일 수 있고, 바람직하게는 락토바실러스 플랜타룸(Lactobacillus plantarum)일 수 있고, 가장 바람직하게는 KCTC 3108로 기탁된 락토바실러스 플랜타룸일 수 있으나, 이에 제한되는 것은 아니다.The term "lactic acid bacteria" refers to living bacteria that reach the intestines, produce lactic acid, and change acidity so that beneficial bacteria can increase. The lactic acid bacteria are, for example, Lactobacillus plantarum , Enterococcus faecium, Lactobacillus pentosus , Lactobacillus acidophilus, Lactobacillus reuteri ( Lactobacillus reuteri ), Lactobacillus buchneri, and Bacillus coagulans, may be at least one selected from the group consisting of, preferably Lactobacillus plantarum , and most preferably KCTC. It may be Lactobacillus plantarum deposited as 3108, but is not limited thereto.
상기 유산균은 활성화된 균주를 사용할 수 있으나, 이에 제한되는 것은 아니다. 상기 균주의 활성화는 활성 배지에 균주를 접종하고 배양하여 수행할 수 있으며, 바람직하게는 식물성 배지에 균주를 접종하고, 37℃에서 24시간 동안 배양할 수 있으나, 이에 제한되는 것은 아니다. 일 구체예에서는, 식물성 배지(dextrose 2%, yeast extract 2%)에 접종하여 1.9 x 109 CFU/mL까지 1차 배양 후, 상기 1차 배양액을 식물성 배지(dextrose 2%, yeast extract 2%)접종하여 2차 배양한 락토바실러스 플랜타룸을 발효에 사용하였다. The lactic acid bacteria may use an activated strain, but is not limited thereto. Activation of the strain may be performed by inoculating and culturing the strain in an active medium, preferably inoculating the strain in a vegetable medium and incubating at 37° C. for 24 hours, but is not limited thereto. In one embodiment, after primary culture up to 1.9 x 10 9 CFU/mL by inoculation in a vegetable medium (dextrose 2%, yeast extract 2%), the primary culture medium was inoculated into a vegetable medium (dextrose 2%, yeast extract 2%) The inoculated and secondary cultured Lactobacillus plantarum was used for fermentation.
녹차, 생강 및 편백의 단독 발효 추출물 제조 시에는, 녹차 추출물, 생강 추출물 또는 편백 추출물 총 중량에 대하여 락토바실러스 플랜타룸을 1 중량% 내지 10 중량%, 바람직하게는 2 중량% 내지 6 중량%, 보다 바람직하게는 2.5 중량% 내지 5.5 중량%, 가장 바람직하게는 3 중량% 내지 5 중량%로 접종하여 녹차 단독 발효 추출물, 생강 단독 발효 추출물 또는 편백 단독 발효 추출물을 제조할 수 있으나, 이에 제한되는 것은 아니다.When preparing the single fermented extract of green tea, ginger and cypress, 1% to 10% by weight of Lactobacillus plantarum, preferably 2% to 6% by weight, based on the total weight of the green tea extract, ginger extract or cypress extract. It is preferably inoculated at 2.5% by weight to 5.5% by weight, most preferably 3% by weight to 5% by weight to prepare a green tea alone fermented extract, a ginger only fermented extract or a cypress alone fermented extract, but is not limited thereto. .
발효 공정은 예를 들어 온도 5℃이상, 6℃이상 또는 7℃이상, 8℃이상, 9℃이상, 또는 10℃이상이고, 55℃이하, 54℃이하, 53℃이하, 52℃이하, 51℃이하, 50℃이하, 49℃이하, 48℃이하, 47℃이하, 46℃이하, 45℃이하, 44℃이하, 43℃이하, 42℃이하, 41℃이하, 40℃이하, 39℃이하, 38℃이하, 37℃이하, 36℃이하 또는 35℃이하, 예컨대, 5℃, 6℃, 7℃, 8℃, 9℃, 10℃, 11℃, 12℃, 13℃, 14℃, 15℃, 16℃, 17℃, 18℃, 19℃, 20℃, 21℃, 22℃, 23℃, 24℃, 25℃, 26℃, 27℃, 28℃, 29℃, 30℃, 31℃, 32℃, 33℃, 34℃, 35℃, 36℃, 37℃, 38℃, 39℃, 40℃, 41℃, 42℃, 43℃, 44℃, 45℃, 46℃, 47℃, 48℃, 49℃, 50℃, 51℃, 52℃, 53℃, 54℃ 또는 55℃, 예컨대 7℃~55℃에서 수행될 수 있고, 바람직하게는 20℃~40℃에서 수행될 수 있고, 보다 바람직하게는 25℃~35℃에서 수행될 수 있다. 온도가 상기 범위를 초과하여 고온이면 유효성분이 파괴될 수 있고, 온도가 상기 범위 미만의 저온이면 반응 시간이 길어지거나 발효가 일어나지 않을 수 있다.The fermentation process is, for example, a temperature of 5°C or higher, 6°C or higher, or 7°C or higher, 8°C or higher, 9°C or higher, or 10°C or higher, and 55°C or lower, 54°C or lower, 53°C or lower, 52°C or lower, 51°C or lower. °C or less, 50 °C or less, 49 °C or less, 48 °C or less, 47 °C or less, 46 °C or less, 45 °C or less, 44 °C or less, 43 °C or less, 42 °C or less, 41 °C or less, 40 °C or less, 39 °C or less . ℃, 16 ℃, 17 ℃, 18 ℃, 19 ℃, 20 ℃, 21 ℃, 22 ℃, 23 ℃, 24 ℃, 25 ℃, 26 ℃, 27 ℃, 28 ℃, 29 ℃, 30 ℃, 31 ℃, 32℃, 33℃, 34℃, 35℃, 36℃, 37℃, 38℃, 39℃, 40℃, 41℃, 42℃, 43℃, 44℃, 45℃, 46℃, 47℃, 48℃ , 49 ° C, 50 ° C, 51 ° C, 52 ° C, 53 ° C, 54 ° C or 55 ° C, such as 7 ° C to 55 ° C, preferably 20 ° C to 40 ° C, more preferably Preferably it may be carried out at 25 ℃ ~ 35 ℃. If the temperature is higher than the above range, the active ingredient may be destroyed, and if the temperature is lower than the above range, the reaction time may be prolonged or fermentation may not occur.
또한, 발효 공정은 예를 들어 15시간 이상, 16시간 이상, 17시간 이상, 18시간 이상, 19시간 이상, 20시간 이상, 21시간 이상, 22시간 이상, 23시간 이상 또는 24시간 이상, 1000시간 이하, 990시간 이하 980시간 이하, 970시간 이하, 960시간 이하, 950시간 이하, 940시간 이하, 930시간 이하, 920시간 이하, 910시간 이하 또는 900시간 이하, 예컨대 15시간 내지 15일 동안 수행할 수 있고, 바람직하게는 20시간 내지 2일 동안 수행할 수 있고, 보다 바람직하게는 24시간 동안 수행할 수 있다.In addition, the fermentation process is, for example, 15 hours or more, 16 hours or more, 17 hours or more, 18 hours or more, 19 hours or more, 20 hours or more, 21 hours or more, 22 hours or more, 23 hours or more or 24 hours or more, 1000 hours 990 hours or less, 980 hours or less, 970 hours or less, 960 hours or less, 950 hours or less, 940 hours or less, 930 hours or less, 920 hours or less, 910 hours or less, or 900 hours or less, such as 15 hours to 15 days. It can be carried out, preferably for 20 hours to 2 days, more preferably for 24 hours.
일 구체예에서는, 락토바실러스 플랜타룸을 녹차 추출물에 대하여 3 중량% 내지 5 중량%, 생강 추출물에 대하여 3 중량% 내지 5 중량%, 편백 추출물에 대하여 3 중량% 내지 5 중량%로 접종하고 30℃에서 24시간 동안 발효시켜 단독 발효 추출물을 제조하였다.In one embodiment, the Lactobacillus plantarum is inoculated at 3% to 5% by weight relative to green tea extract, 3% to 5% by weight relative to ginger extract, and 3% to 5% by weight relative to cypress extract and 30 ° C Fermented for 24 hours to prepare a single fermented extract.
본 명세서에서, 용어 “복합 발효 추출물”은 녹차, 생강 및 편백 원물을 특정 비율로 혼합하여 혼합 추출물을 제조한 이후, 상기 혼합 추출물을 발효시켜 제조한 것을 의미한다.In the present specification, the term "complex fermented extract" means prepared by mixing green tea, ginger, and cypress raw materials in a specific ratio to prepare a mixed extract, and then fermenting the mixed extract.
상기 복합 발효 추출물은 녹차, 생강 및 편백 혼합 추출물 총 중량에 대하여 락토바실러스 플랜타룸을 1 중량% 내지 10 중량%, 바람직하게는 2 중량% 내지 6 중량%, 보다 바람직하게는 2.5 중량% 내지 5.5 중량%, 가장 바람직하게는 3 중량% 내지 5 중량%로 접종하여 제조할 수 있으나, 이에 제한되는 것은 아니다.The complex fermented extract contains 1% to 10% by weight of Lactobacillus plantarum, preferably 2% to 6% by weight, more preferably 2.5% to 5.5% by weight of Lactobacillus plantarum based on the total weight of the mixed extract of green tea, ginger and cypress. %, most preferably, it may be prepared by inoculation at 3% to 5% by weight, but is not limited thereto.
상기 녹차, 생강 및 편백 원물은 2 내지 4:1.5 내지 2.5:1.5 내지 2.5의 중량비로 혼합되는 것일 수 있고, 바람직하게는 2.5 내지 3.5: 1.5 내지 2.5:1.5 내지 2.5의 중량비로 혼합되는 것일 수 있고, 보다 바람직하게는 3:1:1의 중량비로 혼합되는 것일 수 있다. 일 구체예에서는, 녹차, 생강 및 편백 원물을 3:1:1의 중량비로 혼합하여 복합물(복합물 3)을 제조하고, 상기 복합물을 초음파 추출한 혼합 추출물(복합 추출물 3)에 락토바실러스 플랜타룸을 접종하여 복합 발효 추출물(복합 발효 추출물 3)을 제조하였다. 또한, 단독 발효 추출물과 비교할 때, 상기 복합 발효 추출물의 우수한 항염증 활성, 피부탄력 증진 및 피부주름 개선 효과를 확인하였다. The green tea, ginger and cypress raw materials may be mixed in a weight ratio of 2 to 4: 1.5 to 2.5: 1.5 to 2.5, preferably 2.5 to 3.5: 1.5 to 2.5: may be mixed in a weight ratio of 1.5 to 2.5, , More preferably, it may be mixed in a weight ratio of 3:1:1. In one embodiment, green tea, ginger and cypress raw materials are mixed in a weight ratio of 3: 1: 1 to prepare a complex (complex 3), and inoculate Lactobacillus plantarum into a mixed extract obtained by ultrasonically extracting the complex (composite extract 3). Thus, a complex fermentation extract (complex fermentation extract 3) was prepared. In addition, when compared with the single fermented extract, the excellent anti-inflammatory activity, skin elasticity enhancement and skin wrinkle improvement effects of the complex fermented extract were confirmed.
각 원물의 혼합비가 상기 기재된 범위를 벗어날 경우 각 원물의 혼합에 따른 시너지 효과가 저하되어 항염증 활성, 피부탄력 증진 또는 피부주름 개선 효과가 저하될 수 있다.If the mixing ratio of each raw material is out of the above-described range, the synergistic effect of mixing each raw material may be reduced, and thus the anti-inflammatory activity, skin elasticity enhancement, or skin wrinkle improvement effect may be reduced.
상기 녹차, 생강 및 편백 혼합 추출물을 발효시킨 복합 발효 추출물은 전체로서, 화장료 조성물 전체 중량에 대하여 0.001 내지 20중량%으로 포함될 수 있다. 만약 상기 화장료 조성물 총 중량에 대하여 상기 복합 발효 추출물을 0.001중량% 미만으로 포함시킬 경우에는 함량이 적기 때문에 충분한 피부상태 개선 효과를 얻지 못할 수 있고, 20중량%를 초과하는 경우에는 그 이상의 함량을 첨가하지 않아도 충분한 효과를 구현할 수 있기 때문에 비경제적이다.The complex fermented extract obtained by fermenting the green tea, ginger and cypress mixed extract may be included in an amount of 0.001 to 20% by weight based on the total weight of the cosmetic composition as a whole. If the complex fermented extract is included in less than 0.001% by weight with respect to the total weight of the cosmetic composition, sufficient skin condition improvement effect may not be obtained because the content is small, and if it exceeds 20% by weight, more than 0.001% by weight is added. It is uneconomical because sufficient effects can be achieved without having to do it.
상기 녹차, 생강 및 편백 혼합 추출물을 발효시킨 복합 발효 추출물을 화장료로 사용하는 경우, 상기 발효 추출물 이외에 통상적인 화장료 베이스 성분들을 포함할 수 있고, 예를 들면, 안정화제, 용해화제, 안료 및 향료와 같은 통상적인 함유제, 그리고 담체를 포함할 수 있다. 또한, 화장료 당업계에서 통상적으로 제조되는 어떠한 제형으로도 제조될 수 있으며, 특별히 한정하는 것은 아니지만, 예를 들어, 화장수, 에센스, 로션, 페이스트, 계면활성제 함유 클렌징, 크림, 팩, 젤, 연고, 파우더, 패취 또는 분무제(스프레이)의 제형을 가질 수 있다.When the composite fermented extract obtained by fermenting the green tea, ginger, and cypress mixture extract is used as a cosmetic, it may include conventional cosmetic base ingredients in addition to the fermented extract, for example, a stabilizer, a solubilizing agent, a pigment and a flavoring agent. The same conventional containing agent, and may include a carrier. In addition, it may be prepared in any formulation commonly prepared in the cosmetic industry, but is not particularly limited, for example, lotion, essence, lotion, paste, surfactant-containing cleansing, cream, pack, gel, ointment, It may have the formulation of a powder, patch or aerosol (spray).
상기 화장료 조성물의 제형이 페이스트, 크림 또는 젤인 경우에는 담체 성분으로서 동물성유, 식물성유, 왁스, 파라핀, 전분, 트라칸트, 셀룰로오스 유도체, 폴리에틸렌글리콜, 실리콘, 벤토나이트, 실리카, 탈크 및 산화아연 등으로 이루어진 군에서 선택되는 하나 이상이 이용될 수 있고, 제형이 화장수, 에센스인 경우에는 담체 성분으로서 용매, 용해화제 또는 유탁화제가 이용되며, 예를 들어, 물, 에탄올, 이소프로판올, 에틸 카보네이트, 에틸 아세테이트, 벤질 알코올, 벤질 벤조에이트, 프로필렌글리콜, 1,3-부틸렌글리콜, 오일, 글리세롤 지방족 에스테르, 폴리에틸렌글리콜, 및 소르비탄의 지방산 에스테르 등으로 이루어진 군에서 선택되는 하나 이상이 이용될 수 있다.When the formulation of the cosmetic composition is a paste, cream or gel, animal oil, vegetable oil, wax, paraffin, starch, tracanth, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc, zinc oxide, etc. At least one selected from the group may be used, and when the formulation is lotion or essence, a solvent, solubilizing agent or emulsifying agent is used as a carrier component, for example, water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, At least one selected from the group consisting of benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, oil, glycerol aliphatic ester, polyethylene glycol, and sorbitan fatty acid ester may be used.
또한, 상기 화장료 조성물의 제형이 파우더 또는 스프레인 경우에는 담체 성분으로서 락토오스, 탈크, 실리카, 알루미늄 히드록시드, 칼슘 실리케이트 및 폴리아미드 파우더 등으로 이루어진 군에서 선택된 하나 이상이 이용될 수 있고, 특히 스프레이인 경우에는 추가적으로 클로로플루오로히드로카본, 프로판/부탄 또는 디메틸 에테르와 같은 추진제를 포함할 수 있다. 또한, 상기 화장료 조성물의 제형이 계면활성제 함유 클렌징인 경우에는 담체 성분으로서 지방족 알코올 설페이트, 지방족 알코올 에테르 설페이트, 설포숙신산 모노에스테르, 이세티오네이트, 이미다졸리늄 유도체, 메틸타우레이트, 사르코시네이트, 지방산 아미드 에테르 설페이트, 알킬아미도베타인, 지방족 알코올, 지방산 글리세리드, 지방산 디에탄올아미드, 식물성유, 라놀린 유도체 및 에톡실화글리세롤지방산 에스테르 등으로 이루어진 군에서 선택된 하나 이상이 이용될 수 있다.In addition, when the formulation of the cosmetic composition is a powder or spray, one or more selected from the group consisting of lactose, talc, silica, aluminum hydroxide, calcium silicate, and polyamide powder may be used as a carrier component, and in particular, spray In the case of , it may additionally contain a propellant such as chlorofluorohydrocarbon, propane/butane or dimethyl ether. In addition, when the formulation of the cosmetic composition is surfactant-containing cleansing, as carrier components, aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyl taurate, sarcosinate, At least one selected from the group consisting of fatty acid amide ether sulfate, alkylamidobetaine, fatty alcohol, fatty acid glyceride, fatty acid diethanolamide, vegetable oil, lanolin derivative, and ethoxylated glycerol fatty acid ester may be used.
다른 양상은 녹차, 생강 및 편백 혼합 추출물을 발효시킨 복합 발효 추출물을 포함하는 피부 상태 개선용 식품 조성물을 제공한다.Another aspect provides a food composition for improving skin conditions comprising a fermented complex fermented extract of green tea, ginger and cypress mixed extract.
상기 녹차, 생강 및 편백 혼합 추출물을 발효시킨 복합 발효 추출물을 식품 첨가물로 사용할 경우, 상기 복합 발효 추출물을 그대로 첨가하거나 다른 식품 또는 식품 성분과 함께 사용될 수 있고, 통상적인 방법에 따라 적절하게 사용될 수 있다. 유효성분의 혼합양은 사용 목적(예방, 건강 또는 치료적 처치)에 따라 적합하게 결정될 수 있다. 일반적으로, 식품 또는 음료의 제조 시에 본 발명의 복합 발효 추출물은 원료에 대하여 15 중량% 이하, 바람직하게는 10 중량% 이하의 양으로 첨가된다. 그러나, 건강 및 위생을 목적으로 하거나 또는 건강 조절을 목적으로 하는 장기간의 섭취의 경우에는 상기 범위 이하일 수 있으며, 안전성 면에서 아무런 문제가 없기 때문에 유효성분은 상기 범위 이상의 양으로도 사용될 수 있다.When the complex fermented extract obtained by fermenting the green tea, ginger and cypress mixed extract is used as a food additive, the complex fermented extract may be added as it is or used together with other foods or food ingredients, and may be appropriately used according to a conventional method. . The mixing amount of the active ingredient may be appropriately determined according to the purpose of use (prevention, health or therapeutic treatment). In general, when producing food or beverage, the complex fermented extract of the present invention is added in an amount of 15% by weight or less, preferably 10% by weight or less, based on the raw material. However, in the case of long-term intake for the purpose of health and hygiene or health control, it may be below the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount above the above range.
상기 식품의 종류에는 특별한 제한은 없다. 상기 물질을 첨가할 수 있는 식품의 예로는 육류, 소시지, 빵, 초콜릿, 캔디류, 스낵류, 과자류, 피자, 라면, 기타 면류, 껌류, 아이스크림류를 포함한 낙농제품, 각종 수프, 음료수, 차, 드링크제, 알코올 음료 및 비타민 복합제 등이 있으며, 통상적인 의미에서의 건강식품을 모두 포함한다.There is no particular limitation on the type of food. Examples of foods to which the substance can be added include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, chewing gum, dairy products including ice cream, various soups, beverages, tea, drinks, There are alcoholic beverages and vitamin complexes, and includes all health foods in a conventional sense.
본 발명의 건강음료 조성물은 통상의 음료와 같이 여러 가지 향미제 또는 천연 탄수화물 등을 추가 성분으로서 포함할 수 있다. 상술한 천연 탄수화물은 포도당, 과당과 같은 모노사카라이드, 말토오스, 수크로오스와 같은 디사카라이드, 및 덱스트린, 사이클로덱스트린과 같은 천연 감미제나, 사카린, 아스파르탐과 같은 합성 감미제 등을 사용할 수 있다. 상기 천연 탄수화물의 비율은 본 발명의 조성물 100 ml 당 일반적으로 약 0.01 내지 10 g, 바람직하게는 약 0.01 내지 0.1 g 이다.The health beverage composition of the present invention may include various flavoring agents or natural carbohydrates as additional components, like conventional beverages. The aforementioned natural carbohydrates may include monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, natural sweeteners such as dextrin and cyclodextrin, and synthetic sweeteners such as saccharin and aspartame. The proportion of the natural carbohydrate is generally about 0.01 to 10 g, preferably about 0.01 to 0.1 g per 100 ml of the composition of the present invention.
상기 외에 본 발명의 조성물은 여러 가지 영양제, 비타민, 전해질, 풍미제, 착색제, 펙트산 및 그의 염, 알긴산 및 그의 염, 유기산, 보호성 콜로이드 증점제, pH 조절제, 안정화제, 방부제, 글리세린, 알코올, 탄산 음료에 사용되는 탄산화제 등을 포함할 수 있다. 그 밖에 본 발명의 조성물은 천연 과일주스, 과일주스 음료 및 야채 음료의 제조를 위한 과육을 포함할 수 있다. 이러한 성분은 독립적으로 또는 조합하여 사용할 수 있다. 이러한 첨가제의 비율은 크게 중요하진 않지만 본 발명의 조성물 100 중량부 당 0.01 내지 0.1 중량부의 범위에서 선택되는 것이 일반적이다.In addition to the above, the composition of the present invention contains various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, carbonation agents used in carbonated beverages; and the like. In addition, the composition of the present invention may include fruit flesh for preparing natural fruit juice, fruit juice beverages, and vegetable beverages. These components may be used independently or in combination. The ratio of these additives is not critical, but is generally selected in the range of 0.01 to 0.1 part by weight per 100 parts by weight of the composition of the present invention.
또 다른 양상은 녹차, 생강 및 편백 혼합 추출물을 발효시킨 복합 발효 추출물을 포함하는 항염증용 약학적 조성물을 제공한다.Another aspect provides an anti-inflammatory pharmaceutical composition comprising a complex fermented extract obtained by fermenting a mixed extract of green tea, ginger and cypress.
상기 약학적 조성물은 염증과 관련된 피부 질환을 예방 또는 치료하기 위한 것일 수 있다. 상기 피부 질환은 피부 염증으로 인한 질환일 수 있고, 예를 들어, 피부염, 아토피 피부염, 알레르기성 피부염, 지루성 피부염, 건선, 여드름 등을 포함할 수 있으나, 이에 제한되는 것은 아니다.The pharmaceutical composition may be for preventing or treating skin diseases associated with inflammation. The skin disease may be a disease caused by skin inflammation, and may include, for example, dermatitis, atopic dermatitis, allergic dermatitis, seborrheic dermatitis, psoriasis, acne, etc., but is not limited thereto.
용어 “예방”은 질병의 발생을 억제하는 것을 포함한다.The term "prevention" includes inhibiting the occurrence of a disease.
용어 “치료”는 질병의 발전의 억제, 경감 또는 제거를 포함한다.The term “treatment” includes the inhibition, alleviation or elimination of the development of a disease.
일 구체예에 따른 약학적 조성물은 NO의 생성을 억제하고, 염증관련인자인 TNF-α 및 IL-6의 발현을 억제하여 피부 염증 질환을 예방 또는 치료할 수 있다. The pharmaceutical composition according to one embodiment can prevent or treat inflammatory skin diseases by inhibiting the production of NO and inhibiting the expression of inflammation-related factors TNF-α and IL-6.
상기 약학적 조성물은, 추가로 약학적으로 허용 가능한 담체를 1종 이상 포함하여 제조할 수 있다. 약학적으로 허용 가능한 담체는 식염수, 멸균수, 링거액, 완충 식염수, 덱스트로오스 용액, 말토덱스트린 용액, 글리세롤, 에탄올 및 이들 성분 중 1 성분 이상을 혼합하여 사용할 수 있으며, 필요에 따라 항산화제, 완충액, 정균제 등 다른 통상의 첨가제를 첨가할 수 있다. 또한, 희석제, 분산제, 계면활성제, 결합제 및 윤활제를 부가적으로 첨가하여 수용액, 현탁액, 유탁액 등과 같은 주사용 제형, 환약, 캡슐, 과립 또는 정제로 제제화할 수 있다. 더 나아가 당 분야의 적정한 방법으로, 각 질환에 따라 또는 성분에 따라 바람직하게 제제화할 수 있다.The pharmaceutical composition may be prepared by further including one or more pharmaceutically acceptable carriers. A pharmaceutically acceptable carrier may be a mixture of saline, sterile water, Ringer's solution, buffered saline, dextrose solution, maltodextrin solution, glycerol, ethanol, and one or more of these components, and, if necessary, antioxidants and buffers. , bacteriostatic agents and other conventional additives may be added. In addition, diluents, dispersants, surfactants, binders, and lubricants may be additionally added to prepare formulations for injections such as aqueous solutions, suspensions, and emulsions, pills, capsules, granules, or tablets. Furthermore, it can be preferably formulated according to each disease or component by an appropriate method in the art.
상기 약학적 조성물은 녹차, 생강 및 편백 혼합 추출물을 발효시킨 복합 발효 추출물과 함께 항염증 효과를 갖는 공지의 유효성분을 1종 이상 함유할 수 있다.The pharmaceutical composition may contain at least one known active ingredient having an anti-inflammatory effect together with a complex fermented extract obtained by fermenting a mixed extract of green tea, ginger and cypress.
또 다른 양상은 상기 항염증용 약학적 조성물을 개체에 투여하는 단계를 포함하는 염증을 치료하는 방법을 제공한다.Another aspect provides a method for treating inflammation comprising administering the anti-inflammatory pharmaceutical composition to a subject.
용어 “투여”는 임의의 적절한 방법으로 개체에게 약학적 조성물을 제공하는 것을 의미한다.The term “administering” means providing a pharmaceutical composition to a subject by any suitable method.
상기 개체는 인간 또는 비-인간을 포함하는 포유류이며, 비-인간 포유류는 마우스, 랫트, 개, 고양이, 말, 소, 양, 염소, 돼지, 토끼 등을 포함하나 이에 제한되는 것은 아니다.The subject is a mammal including a human or a non-human, and the non-human mammal includes, but is not limited to, mouse, rat, dog, cat, horse, cow, sheep, goat, pig, rabbit, and the like.
상기 약학적 조성물은 목적하는 방법에 따라 경구 투여하거나 비경구 투여(예를 들어, 정맥 내, 피하, 복강 내 또는 국소에 적용)할 수 있으며, 투여량은 환자의 체중, 연령, 성별, 건강상태, 식이, 투여시간, 투여방법, 배설율 및 질환의 중증도 등에 따라 그 범위가 다양하다. 상기 조성물의 일일 투여량은 약 0.0001~500 mg/kg, 바람직하게는 약 0.001~300 mg/kg이며, 하루 일회 내지 수회에 나누어 투여하는 것이 바람직하다.The pharmaceutical composition may be administered orally or parenterally (for example, intravenously, subcutaneously, intraperitoneally or topically applied) according to the desired method, and the dosage is determined according to the patient's weight, age, sex, and health condition. , the range varies depending on the diet, administration time, administration method, excretion rate, and severity of the disease. The daily dose of the composition is about 0.0001 to 500 mg/kg, preferably about 0.001 to 300 mg/kg, and it is preferable to divide the administration once or several times a day.
일 양상에 따른 조성물은 녹차, 생강 및 편백 혼합 추출물을 발효시킨 복합 발효 추출물을 유효성분으로 포함하며, 염증 관련 단백질의 발현 억제를 통한 항염증 효과가 우수하고, 피부탄력을 유지시키는 단백질인 엘라스틴을 분해하는 엘라스타제 활성을 효과적으로 억제하여 피부탄력 증진 및 피부주름 개선 효과를 제공할 수 있다. 상기 조성물은 천연물을 이용하여 안전하고, 기존 의약품에 비해 장기간 사용이 용이하여 화장품, 식품 등 다양한 분야에서 유용하게 활용될 수 있다.The composition according to one aspect includes, as an active ingredient, a complex fermented extract obtained by fermenting a mixed extract of green tea, ginger, and cypress, and has an excellent anti-inflammatory effect by suppressing the expression of inflammation-related proteins and elastin, a protein that maintains skin elasticity. By effectively suppressing the decomposing elastase activity, it is possible to provide skin elasticity enhancement and skin wrinkle improvement effects. The composition is safe by using natural products, and can be usefully used in various fields such as cosmetics and food because it is easy to use for a long period of time compared to existing pharmaceuticals.
도 1 및 2는 단독 추출물의 항염 및 피부탄력 증진 효과를 확인한 결과를 나타낸 도이다.
도 3 및 4는 복합 추출물의 항염 및 피부탄력 증진 효과를 확인한 결과를 나타낸 도이다.
도 5 및 6은 단독 발효 추출물의 항염 및 피부탄력 증진 효과를 확인한 결과를 나타낸 도이다.
도 7 및 8은 복합 발효 추출물의 항염 및 피부탄력 증진 효과를 확인한 결과를 나타낸 도이다.1 and 2 are diagrams showing the results of confirming the anti-inflammatory and skin elasticity enhancing effects of a single extract.
3 and 4 are diagrams showing the results of confirming the anti-inflammatory and skin elasticity enhancing effects of the complex extract.
5 and 6 are diagrams showing the results of confirming the anti-inflammatory and skin elasticity enhancing effects of the single fermented extract.
7 and 8 are diagrams showing the results of confirming the anti-inflammatory and skin elasticity enhancing effects of the complex fermented extract.
이하, 실시예를 통하여 본 발명을 더욱 상세히 설명하고자 한다. 이들 실시예는 오로지 본 발명을 예시하기 위한 것으로서, 본 발명의 범위가 이들 실시예에 의해 제한되는 것으로 해석되지는 않는 것은 당업계에서 통상의 지식을 가진 자에게 있어서 자명할 것이다.Hereinafter, the present invention will be described in more detail through examples. These examples are only for illustrating the present invention, and it will be apparent to those skilled in the art that the scope of the present invention is not to be construed as being limited by these examples.
제조예 1. 단독 추출물의 제조Preparation Example 1. Preparation of single extract
녹차잎(제주자원식물연구소) 및 편백잎(자연초)은 물로 세척하여 이물질을 제거한 후 건조하였다. 생강(동광한방몰)은 뿌리줄기 말린 것을 세척 후 건조한 것을 절편 모양으로 잘라 준비하였다. 편백잎, 녹차잎 및 생강 절편을 각각 50g으로 준비하고, 각 원물을 30%(v/v) 부틸렌글리콜 수용액과 1:15의 중량 비율로 혼합하여 소니케이터(sonicator)에 넣은 다음, 25 KHz, 20℃ 조건에서 1시간 동안 3회 초음파 추출하였다. 추출 후 여과지(0.2 um, Millipore membrane filter, 미국)로 여과하여 얻은 단독 추출물을 이후 실험에 사용하였다. Green tea leaves (Jeju Plant Resources Research Institute) and cypress leaves (natural herb) were washed with water to remove foreign substances and then dried. Ginger (Donggwang Oriental Medicine Mall) was prepared by washing dried rhizomes and cutting the dried ones into slices. Cypress leaves, green tea leaves, and ginger slices were prepared in 50 g each, and each raw material was mixed with 30% (v / v) butylene glycol aqueous solution in a weight ratio of 1: 15, put in a sonicator, and then 25 Ultrasonic extraction was performed three times for 1 hour at KHz and 20°C. After extraction, the single extract obtained by filtering with filter paper (0.2 um, Millipore membrane filter, USA) was used in subsequent experiments.
제조예 2. 복합 추출물의 제조Preparation Example 2. Preparation of composite extract
각 원물을 다음과 같은 중량비로 혼합하여 준비하였다:Each raw material was prepared by mixing in the following weight ratio:
- 녹차:생강:편백 = 2:1.5:1.5 (복합물 1)- Green tea: ginger: cypress = 2:1.5:1.5 (complex 1)
- 녹차:생강:편백 = 2:1:2 (복합물 2)- Green tea: ginger: cypress = 2:1:2 (complex 2)
- 녹차:생강:편백 = 3:1:1 (복합물 3)- Green tea: ginger: cypress = 3:1:1 (complex 3)
상기 원물 혼합물을 30%(v/v) 부틸렌글리콜 수용액과 1:15의 중량 비율로 혼합하여 소니케이터(sonicator)에 넣은 다음, 25 KHz, 20℃ 조건에서 1시간 동안 3회 초음파 추출하였다. 추출 후 여과지(0.2 um, Millipore membrane filter)로 여과하여 얻은 복합 추출물 1 내지 3을 이후 실험에 사용하였다.The original mixture was mixed with a 30% (v / v) butylene glycol aqueous solution at a weight ratio of 1:15, put into a sonicator, and then ultrasonically extracted three times for 1 hour at 25 KHz and 20 ° C. . After extraction, the
제조예 3. 발효 추출물의 제조Preparation Example 3. Preparation of fermented extract
3-1. 사용 균주3-1. Strains used
한국생명공학연구원 미생물자원센터(KCTC)에서 분양 받은 락토바실러스 플랜타룸(Lactobacillus plantarum) KCTC 3108 균주를 사용하였다. 냉장고에 있는 스탁(stock) 균주를 식물성 액체배지(dextrose 2%, yeast extract 2%) 또는 고체배지(dextrose 2%, yeast extract 2%, agar 1.5%)에 접종하여 배양하였다. Lactobacillus plantarum KCTC 3108 strain obtained from the Microbial Resource Center (KCTC) of the Korea Research Institute of Bioscience and Biotechnology was used. The stock strain in the refrigerator was inoculated into vegetable liquid medium (dextrose 2%, yeast extract 2%) or solid medium (dextrose 2%, yeast extract 2%, agar 1.5%) and cultured.
고체배지에서 배양한 단일 콜로니를 취하여 식물성 액체배지에 접종하여 37℃에서 24시간 동안 1.9 x 108 CFU/ml까지 1차 배양하였다(pH 3.80). 상기 1차 배양액(100ml)을 식물성 배지에 3%(v/v)로 접종하여 37℃에서 24시간 동안 2차 배양하였다(pH 3.82). 2차 배양이 끝난 후 121℃에서 15분간 멸균시킨 뒤, 여과지(0.2 um, Millipore membrane filter)로 여과하여 이후 실험에 사용하였다.A single colony cultured on a solid medium was inoculated into a vegetable liquid medium and primary cultured up to 1.9 x 10 8 CFU/ml at 37° C. for 24 hours (pH 3.80). The primary culture medium (100ml) was inoculated in a vegetable medium at 3% (v/v) and secondary culture was performed at 37° C. for 24 hours (pH 3.82). After the secondary culture was completed, the samples were sterilized at 121° C. for 15 minutes, and then filtered with a filter paper (0.2 um, Millipore membrane filter) to be used in subsequent experiments.
3-2. 발효 추출물의 제조3-2. Preparation of fermented extract
상기 제조한 락토바실러스 플랜타룸 배양 여과액을 이용하여, 제조예 1에서 제조한 단독 추출물의 유산균 발효물을 제조하였다. 구체적으로, 상기 락토바실러스 플랜타룸 배양 여과액을 녹차 단독 추출물에 대하여 3 - 5 중량%, 생강 단독 추출물에 대하여 3 - 5 중량%, 편백 단독 추출물에 대하여 3 - 5 중량%로 접종하고 30℃에서 24시간 동안 발효시켰다. 만들어진 각 발효 추출물을 여과지(0.2 um, Millipore membrane filter)로 여과하고, 잡균의 생장을 억제하기 위해 121℃ 이상에서 15분 내지 20분간 동안 가열하여 멸균하였다. 동일한 방법으로, 상기 락토바실러스 플랜타룸 배양 여과액을 제조예 2에서 제조한 복합 추출물 3에 대하여 3-5 중량%로 접종하여 복합 발효 추출물 3을 제조하였다.A lactic acid bacteria fermentation product of the single extract prepared in Preparation Example 1 was prepared using the prepared Lactobacillus plantarum culture filtrate. Specifically, the Lactobacillus plantarum culture filtrate was inoculated at 3 - 5% by weight relative to green tea alone extract, 3 - 5% by weight relative to ginger extract alone, and 3 - 5% by weight relative to cypress alone extract, and at 30 ° C. Fermented for 24 hours. Each prepared fermented extract was filtered with a filter paper (0.2 um, Millipore membrane filter), and sterilized by heating at 121 ° C. or higher for 15 to 20 minutes to inhibit the growth of germs. In the same way, the complex fermented extract 3 was prepared by inoculating the Lactobacillus plantarum culture filtrate at 3-5% by weight with respect to the complex extract 3 prepared in Preparation Example 2.
실험예 1. 단독 추출물의 항염, 피부상태 개선 효과 확인Experimental Example 1. Confirmation of anti-inflammatory and skin condition improvement effect of single extract
상기 제조예 1에서 제조한 단독 추출물의 항염 및 피부상태 개선(피부탄력 증진, 피부주름 개선) 효과를 평가하기 위해, 산화질소(NO) 생성 억제 효과 및 엘라스타제(elastase) 저해 활성을 측정하였다. NO의 양은 안정한 산화 산물인 아질산염의 양을 측정함으로써 확인하였다. 구체적으로, RAW 264.7 세포(1x104 cells/웰)를 96-웰 플레이트에서 12-18 시간 동안 배양한 후, 무혈청 상태에서 1 ㎍/㎖의 LPS를 처리하였다. 이후, 1%(w/v), 5%(w/v) 및 10%(w/v)의 단독 추출물을 함유하는 배지에서 24시간 동안 배양하였고, 세포 배양 상등액 100 ㎕를 새로운 96-웰 플레이트에서 그리스 시약(Sigma-Aldrich Co., St. Louis, MO, USA) 100 ㎕와 혼합한 후, 550 nm에서 흡광도를 측정하였다. 아질산염 농도는 아질산 나트륨 표준 곡선과 비교하여 결정하였다. 양성 대조군으로는 아스코르브산(ascorbic acid)을 사용하였다.In order to evaluate the anti-inflammatory and skin condition improvement (skin elasticity enhancement, skin wrinkle improvement) effect of the single extract prepared in Preparation Example 1, the nitric oxide (NO) production inhibitory effect and elastase inhibitory activity were measured. . The amount of NO was confirmed by measuring the amount of nitrite, a stable oxidation product. Specifically, after culturing RAW 264.7 cells (1x10 4 cells/well) in a 96-well plate for 12-18 hours, they were treated with 1 μg/ml of LPS in a serum-free state. Thereafter, culture was performed for 24 hours in a medium containing 1% (w/v), 5% (w/v), and 10% (w/v) of the extract alone, and 100 μl of the cell culture supernatant was added to a new 96-well plate. After mixing with 100 μl of grease reagent (Sigma-Aldrich Co., St. Louis, MO, USA), the absorbance was measured at 550 nm. Nitrite concentration was determined by comparison to a sodium nitrite standard curve. As a positive control, ascorbic acid was used.
다음으로, 엘라스타제 저해 활성을 측정하기 위해 피부 섬유아세포인 HDF-N 세포에 0.1% triton X-100 이 함유된 0.2M Tris-HCl(pH 8.0)을 첨가한 후 초음파 분쇄기로 균질화시켰다. 이를 3,000rpm에서 20분 동안 원심분리하여 얻은 상층액을 엘라스타제 저해 활성 측정을 위한 시료로 사용하였다. 상기 시료 200 ㎍/ml, 0.2M Tris-HCl 완충액 및 1%(w/v), 5%(w/v) 및 10%(w/v)의 단독 추출물을 농도 별로 각각 혼합한 후, 50 mM N-STANA를 넣고 37℃에서 배양한 후 405 nm에서 흡광도를 측정하였다. 양성 대조군으로는 EGCG(epigallocatechin gallate)를 사용하였다. 그 결과를 도 1 내지 2 및 하기 표 1 내지 2에 나타내었다.Next, in order to measure the elastase inhibitory activity, 0.2M Tris-HCl (pH 8.0) containing 0.1% triton X-100 was added to HDF-N cells, which are dermal fibroblasts, and homogenized using an ultrasonicator. The supernatant obtained by centrifugation at 3,000 rpm for 20 minutes was used as a sample for measuring elastase inhibitory activity. 200 μg/ml of the sample, 0.2M Tris-HCl buffer, and 1% (w/v), 5% (w/v), and 10% (w/v) single extract were mixed by concentration, respectively, and 50 mM N-STANA was added and incubated at 37° C., and absorbance was measured at 405 nm. EGCG (epigallocatechin gallate) was used as a positive control. The results are shown in Figures 1 and 2 and Tables 1 and 2 below.
도 1 내지 2, 표 1 내지 2에 나타낸 바와 같이, 녹차, 생강 및 편백 단독 추출물은 농도의존적인 항염 활성을 나타내었고, 녹차 및 편백 단독 추출물은 농도의존적인 엘라스타제 저해 활성을 나타냄을 확인하였다.As shown in Figures 1 and 2 and Tables 1 and 2, it was confirmed that the extracts of green tea, ginger and cypress alone showed concentration-dependent anti-inflammatory activity, and the extracts of green tea and cypress alone showed concentration-dependent elastase inhibitory activity. .
실험예 2. 복합 추출물의 항염, 피부상태 개선 효과 확인Experimental Example 2. Confirmation of anti-inflammatory and skin condition improvement effects of complex extracts
상기 제조예 2에서 제조한 복합 추출물의 항염 및 피부상태 개선 효과를 평가하기 위해, 산화질소(NO) 생성 억제 효과 및 엘라스타제(elastase) 저해 활성을 측정하였다. 실험은 상기 실험예 1에 기재한 방법과 동일한 방법으로 수행하였고, 그 결과를 도 3 내지 4 및 하기 표 3 내지 4에 나타내었다.In order to evaluate the anti-inflammatory and skin condition improving effects of the composite extract prepared in Preparation Example 2, nitric oxide (NO) production inhibitory effect and elastase inhibitory activity were measured. The experiment was performed in the same manner as described in Experimental Example 1, and the results are shown in FIGS. 3 to 4 and Tables 3 to 4 below.
도 3 내지 4, 표 3 내지 4에 나타낸 바와 같이, 복합 추출물 1 내지 3 모두 농도의존적인 항염 및 엘라스타제 저해 활성을 나타냄을 확인하였다. 특히, 녹차:생강:편백=3:1:1의 중량비로 혼합하여 제조한 복합 추출물 3의 항염 및 엘라스타제 저해 활성이 복합 추출물 1 및 2와 비교할 때 2배 이상의 높은 활성을 보임을 확인하여, 추후 복합 발효 추출물 제조에 사용하였다.As shown in Figures 3 to 4 and Tables 3 to 4, it was confirmed that all of the
실험예 3. 단독 발효 추출물의 항염, 피부상태 개선 효과 확인Experimental Example 3. Confirmation of anti-inflammatory and skin condition improvement effects of single fermented extract
상기 제조예 3-2에서 제조한 단독 발효 추출물의 항염 및 피부상태 개선 효과를 평가하였다. 실험은 상기 실험예 1에 기재한 방법과 동일한 방법으로 수행하였고, 그 결과를 도 5 내지 6 및 하기 표 5 내지 6에 나타내었다.The anti-inflammatory and skin condition improvement effects of the single fermented extract prepared in Preparation Example 3-2 were evaluated. The experiment was performed in the same manner as described in Experimental Example 1, and the results are shown in FIGS. 5 to 6 and Tables 5 to 6 below.
도 5 내지 6, 표 5 내지 6에 나타낸 바와 같이, 녹차, 생강 및 편백 단독 발효 추출물은 모두 농도의존적인 항염 활성을 나타내었고, 녹차 및 편백 단독 발효 추출물은 농도의존적인 엘라스타제 저해 활성을 나타냄을 확인하였다. 한편, 생강 단독 발효 추출물의 경우 엘라스타제 저해 활성 정도가 미미하였다.As shown in Figures 5 to 6 and Tables 5 to 6, green tea, ginger and cypress alone fermented extracts all showed concentration-dependent anti-inflammatory activity, and green tea and cypress alone fermented extracts showed concentration-dependent elastase inhibitory activity confirmed. On the other hand, in the case of the fermented extract of ginger alone, the degree of elastase inhibitory activity was insignificant.
실험예 4. 복합 발효 추출물의 항염, 피부상태 개선 효과 확인Experimental Example 4. Confirmation of anti-inflammatory and skin condition improvement effects of complex fermented extracts
상기 제조예 3-2에서 제조한 복합 발효 추출물 3의 항염 및 피부상태 개선 효과를 평가하였다. 실험은 상기 실험예 1에 기재한 방법과 동일한 방법으로 수행하였고, 그 결과를 도 7 내지 8 및 하기 표 7 내지 8에 나타내었다.The anti-inflammatory and skin condition improvement effects of the complex fermented extract 3 prepared in Preparation Example 3-2 were evaluated. The experiment was performed in the same manner as described in Experimental Example 1, and the results are shown in FIGS. 7 to 8 and Tables 7 to 8 below.
도 7 내지 8, 표 7 내지 8에 나타낸 바와 같이, 복합 발효 추출물 3은 농도의존적인 항염 및 엘라스타제 저해 활성을 나타내며, 단독 발효 추출물에 비해 항염 뿐만 아니라 엘라스타제 저해 활성도 현저히 우수함을 확인하였다.As shown in Figures 7 to 8 and Tables 7 to 8, the complex fermented extract 3 exhibited concentration-dependent anti-inflammatory and elastase inhibitory activities, and compared to the single fermented extract, it was confirmed that the anti-inflammatory as well as the elastase inhibitory activity were significantly superior. .
상기 결과를 통해, 복합 발효 추출물 3은 단독 발효 추출물 중 항염 및 엘라스타제 저해 활성이 가장 우수한 녹차 원물을 가장 높은 비율로 포함하고, 생강 및 편백 원물을 최적 비율로 포함하는(녹차:생강:편백 = 3:1:1) 녹차, 생강 및 편백 혼합 추출물을 발효시켜 제조함으로써, 단독 발효 추출물에 비해 항염 및 엘라스타제 저해 활성이 현저히 증가되어 항염 용도, 피부탄력 증진 및 피부주름 개선 용도로 유용하게 사용할 수 있음을 확인하였다. 뿐만 아니라, 상기 복합 발효 추출물 3은 1%(w/v)의 낮은 농도에서도 현저히 높은 엘라스타제 저해 활성을 나타냄을 확인하였다. Through the above results, the complex fermented extract 3 contains the highest ratio of green tea raw material having the highest anti-inflammatory and elastase inhibitory activity among the single fermented extracts, and contains ginger and cypress raw material at an optimal ratio (green tea: ginger: cypress). = 3:1:1) Made by fermenting a mixed extract of green tea, ginger, and cypress, the anti-inflammatory and elastase inhibitory activities are significantly increased compared to the single fermented extract, making it useful for anti-inflammatory use, skin elasticity enhancement, and skin wrinkle improvement. Confirmed that it can be used. In addition, it was confirmed that the complex fermented extract 3 exhibits remarkably high elastase inhibitory activity even at a low concentration of 1% (w/v).
실험예 5. 세포 독성 확인Experimental Example 5. Confirmation of cytotoxicity
가장 우수한 항염 및 엘라스타제 저해 활성을 나타낸 복합 발효 추출물 3(이하, 복합 발효 추출물이라 함)에 대하여 추가 실험을 수행하였다. 먼저, 세포 독성을 확인하기 위해, 섬유아세포(HS86)를 10% 소혈청을 첨가한 DMEM(Dulbecco's Modified Eagle's Medium) 배지에 1 x 104개로 접종하고 하루 동안 5% CO2, 37℃에서 배양하였다. 0.1%(w/v), 0.5%(w/v) 및 1%(w/v)의 복합 발효 추출물을 무혈청 배지에 희석하여 처리하였다. 1일간 배양 후 MTT(3-(4,5-dimethylthiazol-2-yl)-2,5- diphenyltetrazolium bromide) 시약을 처리하고 4시간 동안 더 배양하였다. 형성된 포마르잔 결정(formazan crystal)을 DMSO에 녹인 후 96-웰 플레이트에 200㎕씩 넣고 540 ㎚에서 ELISA plate reader로 흡광도를 측정하였다. 대조군으로는 복합 발효 추출물을 처리하지 않은 배양액을 사용하였다.Further experiments were performed on the complex fermented extract 3 (hereinafter referred to as the complexed fermented extract) that showed the most excellent anti-inflammatory and elastase inhibitory activities. First, to confirm cytotoxicity, fibroblasts (HS86) were inoculated at 1 x 10 4 in DMEM (Dulbecco's Modified Eagle's Medium) medium supplemented with 10% bovine serum and cultured at 5% CO 2 and 37°C for one day. . 0.1% (w/v), 0.5% (w/v) and 1% (w/v) of the complex fermentation extract were diluted in serum-free medium and treated. After culturing for 1 day, the cells were treated with MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) reagent and further incubated for 4 hours. After dissolving the formed formazan crystals in DMSO, 200 μl of each was placed in a 96-well plate, and absorbance was measured at 540 nm with an ELISA plate reader. As a control, a culture medium not treated with the complex fermentation extract was used.
[식 1][Equation 1]
세포생존율(%) = 100 - (복합 발효 추출물을 처리한 실험군의 흡광도)/(대조군의 흡광도) X 100Cell viability (%) = 100 - (absorbance of the experimental group treated with complex fermented extract) / (absorbance of the control group)
상기 식 1을 사용하여 세포생존율(%)을 계산하고, 대조군의 세포생존율(%)을 100이라고 할 때의 상대적인 세포증식효과를 하기 표 9에 나타내었다.The cell viability (%) was calculated using
상기 표 9에 나타낸 바와 같이, 일 양상에 따른 복합 발효 추출물은 세포 독성을 보이지 않아, 피부에 안정적으로 사용할 수 있음을 확인하였다.As shown in Table 9, it was confirmed that the complex fermented extract according to one aspect did not show cytotoxicity and could be stably used on the skin.
실험예 6. 염증인자 발현 확인Experimental Example 6. Confirmation of expression of inflammatory factors
다음으로, 일 양상에 따른 복합 발효 추출물의 항염 활성을 추가적으로 검증하기 위해 염증인자인 TNF-α 및 IL-6의 발현 억제능을 평가하였다. 구체적으로, 섬유아세포(HS86)를 48-웰 플레이트에 접종하여 24시간 동안 배양하였다. 이후 배지를 교체하고 LPS를 처리한 후, 복합 추출물 3과 복합 발효 추출물을 각각 0.1%(w/v), 0.5%(w/v) 및 1%(w/v)의 농도로 처리한 후 48시간 동안 추가로 배양하였다. 양성대조군으로는 덱사메타손(dexamethasone)을 동일 농도로 처리하였다. 아무 처리도 하지 않은 배양액과 비교하여, 상기 복합 추출물 또는 복합 발효 추출물을 처리한 배양액의 TNF-α 및 IL-6 농도의 상대값을 계산하였다. 상기 TNF-α 및 IL-6의 농도는 ELISA 분석법으로 측정하였다. TNF-α 및 IL-6의 억제율(%)에 대한 결과값을 각각 하기 표 10 및 표 11에 나타내었다.Next, in order to further verify the anti-inflammatory activity of the complex fermented extract according to one aspect, the ability to inhibit the expression of TNF-α and IL-6, which are inflammatory factors, was evaluated. Specifically, fibroblasts (HS86) were inoculated into a 48-well plate and cultured for 24 hours. Then, after replacing the medium and treating with LPS, complex extract 3 and complex fermented extract were treated at concentrations of 0.1% (w/v), 0.5% (w/v), and 1% (w/v), respectively, and then 48 Incubated for an additional hour. As a positive control group, dexamethasone was treated at the same concentration. Compared to the culture medium without any treatment, the relative values of TNF-α and IL-6 concentrations of the culture medium treated with the composite extract or the composite ferment extract were calculated. The concentrations of TNF-α and IL-6 were measured by ELISA assay. Results for the inhibition rate (%) of TNF-α and IL-6 are shown in Table 10 and Table 11, respectively.
상기 표 10 및 표 11에 나타낸 바와 같이, 락토바실러스 플랜타룸으로 발효하지 않은 복합 추출물 3에 비해 복합 발효 추출물이 염증인자인 TNF-α 및 IL-6의 발현을 현저히 감소시키는 것을 확인하였다. 특히, 0.5%(w/v) 이상의 농도에서는 양성대조군인 덱사메타손보다 발현 억제율이 높음을 확인하였다. 상기 결과를 통해, 일 양상에 따른 복합 발효 추출물은 강한 항염증 활성을 가짐을 알 수 있었다.As shown in Tables 10 and 11, it was confirmed that the complex fermented extract significantly reduced the expression of the inflammatory factors TNF-α and IL-6 compared to the complex extract 3 that was not fermented with Lactobacillus plantarum. In particular, at a concentration of 0.5% (w / v) or more, it was confirmed that the expression inhibition rate was higher than that of dexamethasone, a positive control group. Through the above results, it was found that the complex fermented extract according to one aspect had strong anti-inflammatory activity.
상기 결과를 통해, 일 양상에 따른 복합 발효 추출물은 NO 생성을 억제하고, TNF-α 및 IL-6의 발현을 현저히 감소시켜 강력한 항염증 효과를 발휘하며, 높은 엘라스타제 저해 활성으로 피부탄력 및 피부주름을 개선할 수 있음을 확인하였다. 또한, 단독 발효 추출물과 복합 발효 추출물 간의 항염 활성, 피부탄력 증진 활성 비교를 통해 상기 효과는 녹차, 생강 및 편백의 최적 혼합비 및 유산균 발효에 따른 상승적 효과에 따른 것임을 알 수 있었다. 따라서, 일 양상에 따른 복합 발효 추출물은 피부 염증을 비롯한 염증성 질환의 예방, 개선 또는 치료를 위한 식품, 화장품 및 의약품 등에 다양하게 활용될 수 있고, 우수한 피부 보습 효과를 제공할 수 있다.Through the above results, the complex fermented extract according to one aspect inhibits NO production, significantly reduces the expression of TNF-α and IL-6, exerts a strong anti-inflammatory effect, and has high elastase inhibitory activity to improve skin elasticity and It was confirmed that skin wrinkles can be improved. In addition, through comparison of anti-inflammatory activity and skin elasticity enhancing activity between the single fermented extract and the complex fermented extract, it was found that the above effects were due to the synergistic effect of the optimal mixing ratio of green tea, ginger and cypress and the fermentation of lactic acid bacteria. Therefore, the complex fermented extract according to one aspect can be used in various ways such as foods, cosmetics, and pharmaceuticals for the prevention, improvement, or treatment of inflammatory diseases including skin inflammation, and can provide excellent skin moisturizing effects.
이하, 제형예를 통하여 본 발명을 더욱 상세히 설명하고자 한다. 제형예는 오로지 본 발명을 예시하기 위한 것으로서, 본 발명의 범위가 제형예에 의해 제한되는 것으로 해석되지 않는다.Hereinafter, the present invention will be described in more detail through formulation examples. Formulation examples are only for exemplifying the present invention, and the scope of the present invention is not construed as being limited by the formulation examples.
제형예 1. 보습 크림의 제조Formulation Example 1. Preparation of Moisturizing Cream
일 양상에 따른 복합 발효 추출물을 포함하는 보습 크림을 하기 표 12에 나타낸 조성 성분 및 조성비에 따라 통상적인 방법으로 제조하였다. Moisturizing cream containing the complex fermented extract according to one aspect was prepared by a conventional method according to the ingredients and composition ratios shown in Table 12 below.
제형예 2. 영양 세럼의 제조Formulation Example 2. Preparation of nutritional serum
일 양상에 따른 복합 발효 추출물을 포함하는 영양 세럼을 하기 표 13에 나타낸 조성 성분 및 조성비에 따라 통상적인 방법으로 제조하였다.Nutrient serum containing the complex fermented extract according to one aspect was prepared by a conventional method according to the ingredients and composition ratios shown in Table 13 below.
실시예 1. 보습감 및 피부상태 개선 효과 평가Example 1. Moisturizing sensation and skin condition improvement effect evaluation
일 양상에 따른 복합 발효 추출물을 포함하는 화장료 조성물의 보습감 및 피부상태 개선 효과를 평가하기 위하여, 상기 제형예에서 제조한 보습 크림 및 영양 세럼을 이용하여 관능시험을 실시하였다.In order to evaluate the moisturizing effect and skin condition improvement effect of the cosmetic composition containing the complex fermented extract according to one aspect, a sensory test was conducted using the moisturizing cream and nutrient serum prepared in the formulation example.
먼저, 발효 과정을 거치지 않은 혼합 추출물을 이용하여, 제형예 1 및 2와 동일한 방법으로 보습 크림 및 영양 세럼을 제조하였다(비교제형예 1 및 2). 25~35세의 여성 40명을 두 그룹으로 나누어, 그룹 1은 제형예 1 및 2에 따른 보습 크림 및 영양 세럼을, 또는 그룹 2는 비교제형예 1 및 2에 따른 보습 크림 및 영양 세럼을 1일 1회 6주간 지속적으로 사용하게 하였다. 관능평가는 피부 도포 후 느껴지는 보습감, 피부탄력 증진 및 피부주름 개선 등의 피부상태 개선 효과에 대하여 매우 우수(5점), 우수(4점), 보통(3점), 나쁨(2점), 매우 나쁨(1점)의 기준으로 수행하였고, 그 결과를 하기 표 14 및 15에 각각 나타내었다.First, moisturizing cream and nutrient serum were prepared in the same manner as Formulation Examples 1 and 2 using mixed extracts that had not undergone fermentation (Comparative Formulation Examples 1 and 2). 40 women aged 25 to 35 were divided into two groups,
(혼합 추출물)Comparative Formulation Example 1
(mixed extract)
(복합 발효 추출물)Formulation example 1
(complex fermented extract)
(혼합 추출물)Comparative Formulation Example 2
(mixed extract)
(복합 발효 추출물)Formulation Example 2
(complex fermented extract)
(혼합 추출물)Comparative Formulation Example 1
(mixed extract)
(복합 발효 추출물)Formulation example 1
(complex fermented extract)
(혼합 추출물)Comparative Formulation Example 2
(mixed extract)
(복합 발효 추출물)Formulation Example 2
(complex fermented extract)
상기 표 14 및 15에 나타낸 바와 같이, 혼합 추출물로 제조한 비교제형예 1 및 2에 비해 상기 혼합 추출물을 락토바실러스 플랜타룸으로 발효시킨 복합 발효 추출물로 제조한 제형예 1 및 2의 보습감, 피부상태 개선 효과에 대한 관능평가 점수가 현저히 높아, 일 양상에 따른 복합 발효 추출물로 제조한 화장료 조성물의 피부탄력 증진 및 피부주름 개선 정도가 매우 우수함을 알 수 있었다.As shown in Tables 14 and 15, compared to Comparative Formulation Examples 1 and 2 prepared with the mixed extract, the moisturizing feeling and skin condition of Formulation Examples 1 and 2 prepared with the complex fermented extract obtained by fermenting the mixed extract with Lactobacillus plantarum The sensory evaluation score for the improvement effect was remarkably high, and it was found that the degree of skin elasticity enhancement and skin wrinkle improvement of the cosmetic composition prepared with the complex fermented extract according to one aspect was very excellent.
전술한 설명은 예시를 위한 것이며, 본 발명이 속하는 기술분야의 통상의 지식을 가진 자는 본 발명의 기술적 사상이나 필수적인 특징을 변경하지 않고서 다른 구체적인 형태로 쉽게 변형이 가능하다는 것을 이해할 수 있을 것이다. 그러므로 이상에서 기술한 실시예들은 모든 면에서 예시적인 것이며 한정적이 아닌 것으로 이해해야만 한다.The above description is for illustrative purposes, and those skilled in the art will understand that it can be easily modified into other specific forms without changing the technical spirit or essential features of the present invention. Therefore, the embodiments described above should be understood as illustrative in all respects and not limiting.
Claims (10)
상기 피부 상태 개선은 항염증, 피부탄력 증진 및 피부주름 개선으로 이루어진 군으로부터 선택되는 하나 이상인 것인 화장료 조성물.A cosmetic composition for improving skin condition comprising a complex fermented extract obtained by fermenting a mixed extract of green tea, ginger and cypress,
The cosmetic composition of claim 1, wherein the skin condition improvement is at least one selected from the group consisting of anti-inflammatory, skin elasticity enhancement, and skin wrinkle improvement.
상기 피부 상태 개선은 항염증, 피부탄력 증진 및 피부주름 개선으로 이루어진 군으로부터 선택되는 하나 이상인 것인 식품 조성물.A food composition for improving skin condition comprising a complex fermented extract obtained by fermenting a mixed extract of green tea, ginger and cypress,
The improvement of the skin condition is one or more selected from the group consisting of anti-inflammatory, skin elasticity promotion and skin wrinkle improvement food composition.
Anti-inflammatory pharmaceutical composition comprising a complex fermented extract obtained by fermenting a mixed extract of green tea, ginger and cypress.
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| KR102103021B1 (en) | 2019-06-19 | 2020-04-22 | 김영자 | Antiatopyic and antivaginitis tissues containing retinispora extracts and a process for the preparation thereof |
| KR102119825B1 (en) | 2019-10-15 | 2020-06-05 | 주식회사 아미코스메틱 | A cosmetic composition for maintaining the balance of microbiom in the skin comprising brown rice, grean tea and dandelion fermentation extracts and methode thereof |
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| KR102103021B1 (en) | 2019-06-19 | 2020-04-22 | 김영자 | Antiatopyic and antivaginitis tissues containing retinispora extracts and a process for the preparation thereof |
| KR102119825B1 (en) | 2019-10-15 | 2020-06-05 | 주식회사 아미코스메틱 | A cosmetic composition for maintaining the balance of microbiom in the skin comprising brown rice, grean tea and dandelion fermentation extracts and methode thereof |
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