KR20200088665A - Compoisition for skin-whitening comprising green tea extract which has modified amounts of ingredients - Google Patents
Compoisition for skin-whitening comprising green tea extract which has modified amounts of ingredients Download PDFInfo
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- KR20200088665A KR20200088665A KR1020190005248A KR20190005248A KR20200088665A KR 20200088665 A KR20200088665 A KR 20200088665A KR 1020190005248 A KR1020190005248 A KR 1020190005248A KR 20190005248 A KR20190005248 A KR 20190005248A KR 20200088665 A KR20200088665 A KR 20200088665A
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Abstract
Description
본 발명은 성분 함량이 변화된 녹차 추출물을 포함하는 피부 미백용 조성물에 관한 것이다.The present invention relates to a composition for skin whitening comprising a green tea extract with a varying component content.
피부는 물리적 및 화학적 자외선 방어 인자를 갖추고, 다양한 광화학 반응에 의한 피부장애를 최소한으로 방지하기 위한 작용 기작을 가진다. 피부의 각질층은 자외선을 반사 및 확산시킴에 의하여 그 에너지를 감쇄시키며, 또한 멜라닌 색소, SOD(Superoxide Dismutase), 그 외의 항산화 성분 등은 피부 내부에 침투한 자외선을 흡수하고, 그 에너지를 감쇄시키거나 자외선이 이차적으로 발생시키는 활성산소를 소거함에 의하여 피부에 대한 장애를 방어한다.The skin has physical and chemical UV protection factors and has a mechanism of action to minimize skin disorders caused by various photochemical reactions. The stratum corneum of the skin attenuates its energy by reflecting and diffusing ultraviolet rays. In addition, melanin pigments, superoxide dismutase (SOD), and other antioxidant components absorb ultraviolet rays penetrating into the skin and attenuate its energy. It protects against skin disorders by eliminating free radicals, which are secondarily generated by ultraviolet rays.
그러나, 상기와 같은 방어인자의 능력을 초과하는 다량의 자외선을 생체가 받게 되거나, 나이가 들어감에 따라 그 능력이 저하되는 경우 여러 가지 피부장애가 일어난다.However, various skin disorders occur when a living body receives a large amount of ultraviolet rays exceeding the ability of the protective factor as described above, or when the ability decreases with age.
피부에는 대식 세포(Macrophage), 랑게르한스 세포(Langerhans cells) 등 생체계에서 면역을 담당하고 있는 여러 가지 세포가 존재한다. 자외선의 조사에 의하여 이러한 세포는 수적인 감소만이 아니고, 기능적으로도 장애를 받게 된다. 피부색 결정 요인 중에서 가장 큰 부분을 차지하는 것이 피부 중 멜라닌의 분포 상태 및 양이다. 멜라닌(Melanin)은 멜라노사이트(Melanocyte)라는 세포에서 생성되는데 이 멜라노사이트에는 티로시나제(Tyrosinase) 등의 효소가 존재하며, 이들이 함께 작용하여, 생체 내에 항상 존재하는 티로신(Tyrosine)이라는 아미노산을 기질로 중합화 산화반응을 함으로 흑갈색의 색소인 멜라닌을 형성하게 된다. 이렇게 형성된 멜라닌은 멜라노사이트의 수지상 돌기를 통하여 케라티노사이트(Keratinocyte)라는 표피 세포로 이동한다. 여기서 멜라닌은 핵 주변에 모자와 같은 구조를 형성하여 자외선으로부터 유전자를 보호하고 자유 라디칼(Free radical)을 제거하여 세포 내 단백질을 보호하는 등 중요한 역할을 하게 된다.There are many cells in the skin that are responsible for immunity in the biological system, such as macrophages and Langerhans cells. These cells are not only reduced numerically, but also functionally impaired by irradiation with ultraviolet rays. The largest part of the skin color deciding factor is the distribution and amount of melanin in the skin. Melanin is produced in cells called melanocytes, and enzymes such as tyrosinase are present in the melanocytes, and they work together to polymerize the amino acid called tyrosine, which is always present in the body, into a substrate. By oxidation oxidation reaction, melanin, a dark brown pigment, is formed. The melanin thus formed moves to the epidermal cells called keratinocytes through the dendritic projections of melanocytes. Here, melanin forms a hat-like structure around the nucleus, which plays an important role in protecting genes from ultraviolet rays and protecting proteins in cells by removing free radicals.
이러한 멜라닌을 분해하는 효소는 생체 내에 존재하지 않으며, 다만 케라티노사이트가 노화되어 제 기능을 다하고 표피에서 떨어져 나갈 때 함께 피부로부터 제거된다. 하지만 멜라닌이 필요 이상으로 많이 생기게 되면 기미나 주근깨, 점 등과 같은 과색소침착증을 유발하여 미용상으로 좋지 않은 결과를 초래한다.The enzyme that breaks down melanin does not exist in the body, but when keratinocytes age and perform their functions, they are removed from the skin together. However, if more melanin is produced than necessary, hyperpigmentation such as freckles, freckles, and spots may be caused, which results in a bad cosmetic effect.
멜라닌 생산에 영향을 미치는 인자는 여러 가지가 알려져 있는데, 자외선에 의하여 일어나는 멜라닌 생산의 항진 그리고 이에 따른 색소 침착이 화장품 분야에서 아주 중요하다. 색소 침착의 예방 목적으로 미백화장품에 배합되는 약제의 기본적인 메커니즘은 티로시나아제(Tyrosinase) 작용 억제, 티로시나아제 생성 억제, 멜라닌 생성 중간체의 억제, 기존 멜라닌의 환원 및 광산화 억제, 멜라닌 배설의 촉진, 및 자외선 커트 등이다.A number of factors affecting melanin production are known, and the improvement of melanin production caused by ultraviolet rays and thus pigmentation are very important in the cosmetic field. For the purpose of preventing pigmentation, the basic mechanisms of drugs formulated in whitening cosmetics are tyrosinase action suppression, tyrosinase production suppression, melanin production intermediate suppression, reduction of existing melanin and photooxidation suppression, promotion of melanin excretion, And UV cuts.
자외선 노출 환경 속에서도 하얀 피부를 갖고자 하는 욕구에 따라서 피부색소 이상 증상과 과색소 침착 등의 예방과 개선에 대한 니즈가 더욱 늘어나고 있으며 이에 과도한 멜라닌 생성을 억제하는 미백 제품 개발이 필요하게 되었고, 그동안 많은 노력들이 이루어졌다. 그 구체적인 예를 들면, 코지산(Kojic acid), 알부틴(Arbutin) 등과 같은 티로시나제 활성을 저해하는 억제제들, 하이드로퀴논(Hydroquinone), 비타민 A, 비타민 C 및 이들의 유도체 등이 있다. 그러나, 이들은 미백 효과의 불충분, 제형 내에서의 안정성 및 피부에 대한 안전성 등의 문제로 인해 그 사용이 제한되고 있다. 이에 대해 피부 미백에 효능이 높으면서도 제형화가 용이하며 피부에 안전한 천연물 유래의 원료들에 대한 연구가 활발히 진행되고 있다.In accordance with the desire to have white skin even in an environment exposed to UV rays, the need for prevention and improvement of skin pigment abnormalities and hyperpigmentation is increasing. Accordingly, it has become necessary to develop a whitening product that suppresses excessive melanin production. Efforts have been made. Specific examples thereof include inhibitors that inhibit tyrosinase activity, such as kojic acid and arbutin, hydroquinone, vitamin A, vitamin C, and derivatives thereof. However, their use is limited due to insufficient whitening effects, stability in the formulation, and safety to the skin. On the other hand, studies on raw materials derived from natural products that have high efficacy in skin whitening and are easy to formulate and are safe for skin are being actively conducted.
본 발명은 독성이 없고 피부 미백 효능이 뛰어난 천연식물 유래의 조성물 및 이의 제조방법을 제공하는 것을 목적으로 한다.An object of the present invention is to provide a composition derived from a natural plant having no toxicity and excellent skin whitening efficacy and a method for manufacturing the same.
상기한 목적을 달성하기 위하여, 본 발명은 일 측면에서, 조성물 총 중량을 기준으로, 4 내지 15 중량%의 (-)-갈로카테킨 갈레이트((-)-gallocatechin gallate, GCG)및 4 내지 15 중량%의 (-)-에피갈로카테킨 갈레이트((-)-epigallocatechin gallate, EGCG)를 포함하는 녹차 추출물을 유효성분으로 포함하는 피부 미백용 조성물을 제공한다.In order to achieve the above object, the present invention, in one aspect, based on the total weight of the composition, 4 to 15% by weight of (-)-gallocatechin gallate ((-)-gallocatechin gallate, GCG) and 4 to 15 Provided is a composition for whitening skin comprising green tea extract as an active ingredient, containing (%)-epigallocatechin gallate (EGCG) by weight (-)-epigallocatechin gallate.
본 발명의 일 측면에 따른 추출물 및 조성물은 피부에 자극이 적고 친환경적인 천연식물 유래의 안전한 것이며, 이를 통해 멜라닌 생성 및 티로시나아제의 활성을 효과적으로 억제하고 우수한 피부 미백 효능을 나타낼 수 있다.The extracts and compositions according to one aspect of the present invention are safe from natural plants with little irritation to the skin and can effectively suppress melanin production and tyrosinase activity and exhibit excellent skin whitening efficacy.
도 1은 실시예 1의 고시형 녹차 추출물(시료 1)에 대한 크로마토그램을 나타낸 것이다.
도 2는 본 발명의 일 측면에 따른 고온처리 녹차 추출물(시료 2)에 대한 크로마토그램을 나타낸 것이다.
도 3은 실험예 1에 따른 고온처리 녹차 추출물의 세포 독성 확인 실험 결과를 나타낸 것이다.
도 4는 실험예 2에 따른 고온처리 녹차 추출물의 멜라닌 세포 생성 억제 효과 실험 결과를 나타낸 것이다.
도 5는 실험예 3에 따른 고온처리 녹차 추출물의 티로시나아제 활성 억제 효과 실험 결과를 나타낸 것이다.FIG. 1 shows a chromatogram for the noticed green tea extract of Example 1 (Sample 1).
Figure 2 shows a chromatogram for the high-temperature treated green tea extract (sample 2) according to an aspect of the present invention.
Figure 3 shows the results of the cytotoxicity test of the high-temperature treated green tea extract according to Experimental Example 1.
Figure 4 shows the results of the melanin cell production inhibitory effect of the high-temperature treated green tea extract according to Experimental Example 2.
Figure 5 shows the experimental results of the inhibitory effect of tyrosinase activity of the high-temperature treated green tea extract according to Experimental Example 3.
본 명세서에서 "녹차 추출물"은 추출방법, 추출용매, 추출된 성분 또는 추출물의 형태를 불문하고, 차과에 속하는 상록수인 차(카멜리아 시넨시스; Camellia sinensis)로부터 추출한 것 또는 바실러스 서브틸리스(Bacillus subtilis spp.)를 접종하고 발효시킨 차잎 등으로부터 추출한 것을 포함하고, 상기 추출한 것을 특정 용매로 분획한 분획물을 포함한다. 상기 차는 차나무 잎, 꽃, 줄기, 열매, 뿌리, 줄기, 및 뿌리의 심재로 구성된 군에서 선택되는 하나 이상이 포함되며, 바람직하게는 잎일 수 있다. 또한, 상기 추출물의 형태는 바람직하게는 분말형태일 수 있다. 상기 추출 또는 분획은 물, 유기 용매, 또는 이들의 혼합용매를 사용하여 할 수 있다. 유기용매는 알코올, 이소프로판올, 아세톤, 헥산, 에틸아세테이트, 이산화탄소, 또는 이들 중 둘 이상의 혼합용매를 사용할 수 있으나 이에 한정되지는 않으며, 녹차의 유효 성분이 파괴되지 않거나 최소화된 조건에서 실온 또는 가온하여 추출 또는 분획할 수 있다. 상기 알코올은 C1~C5의 저급 알코올일 수 있다. 추출 또는 분획의 횟수나 방법은 특별히 제한되지 않으며, 예를 들어 냉침 추출, 초음파 추출, 환류 냉각 추출, 열수 추출 등의 방법을 사용할 수 있고, 바람직하게는 냉침 또는 가온하여 유효성분을 추출 또는 분획하고 여과한 다음, 그 여과액을 감압농축하여 본 발명의 녹차 추출물을 얻을 수 있다.In the present specification, "green tea extract", regardless of the extraction method, the extraction solvent, or the form of the extracted component or extract, is an extract of Bacillus subtilis or Bacillus subtilis , which is an evergreen tree belonging to the family Caraceae ( Camellia sinensis ). spp.), and extracts from fermented tea leaves, etc., and fractions of the extracted ones with a specific solvent. The tea includes one or more selected from the group consisting of tea tree leaves, flowers, stems, fruits, roots, stems, and cores of the roots, and preferably may be leaves. In addition, the form of the extract may be preferably a powder form. The extraction or fractionation can be performed using water, an organic solvent, or a mixed solvent thereof. The organic solvent may use alcohol, isopropanol, acetone, hexane, ethyl acetate, carbon dioxide, or a mixed solvent of two or more of them, but is not limited thereto, and the active ingredient of green tea is not destroyed or extracted by heating at room temperature or in a minimized condition Or fractionation. The alcohol may be a lower alcohol of C 1 ~ C 5 . The number or method of extraction or fractionation is not particularly limited, and for example, methods such as cold immersion extraction, ultrasonic extraction, reflux cooling extraction, and hot water extraction may be used. Preferably, the active ingredient is extracted or fractionated by cold immersion or warming. After filtration, the filtrate is concentrated under reduced pressure to obtain a green tea extract of the present invention.
본 명세서에서 "에피 카테킨"이란, 에피갈로카테킨(epigallocatechin, EGC), (-)에피카테킨((-)epicattechin, EC), (-)-에피갈로카테킨 갈레이트((-)-epigallocatechin gallate, EGCG), 및 에피카테킨 3-O-갈레이트(epicatechin 3-O-gallate, ECG)을 포함하는 것이다.As used herein, "epi catechin" refers to epigallocatechin (EGC), (-) epicatechin ((-)epicattechin, EC), (-)-epigalocatechin gallate ((-)-epigallocatechin gallate, EGCG), and epicatechin 3-O-gallate (ECG).
본 명세서에서 "에피 카테킨 에피머"란, 갈로카테킨 (gallocatechin, GC), 카테킨 (catechin, C), (-)-갈로카테킨 갈레이트 ((-)-gallocatechin gallate, GCG) 및 카테킨 갈레이트 (catechin gallate, CG)를 포함하는 것이다.As used herein, the term "epi catechin epimer" refers to gallocatechin (GC), catechin (C), (-)-gallocatechin gallate (GCG) and catechin gallate (catechin) gallate, CG).
본 발명은 일 측면에서, 추출물 총 중량을 기준으로, 4 내지 15 중량%의 (-)-갈로카테킨 갈레이트((-)-gallocatechin gallate, GCG) 및 4 내지 15 중량%의 (-)-에피갈로카테킨 갈레이트((-)-epigallocatechin gallate, EGCG)를 포함하는 녹차 추출물을 유효성분으로 포함하는 피부 미백용 조성물에 관한 것일 수 있다.In one aspect, the present invention, based on the total weight of the extract, 4 to 15% by weight of (-)-gallocatechin gallate ((-)-gallocatechin gallate, GCG) and 4 to 15% by weight of (-)-epi It may be related to a composition for skin whitening comprising a green tea extract containing gallocatechin gallate ((-)-epigallocatechin gallate, EGCG) as an active ingredient.
일 측면에서 상기 GCG는 상기 추출물 총 중량을 기준으로 4 중량% 이상, 5 중량% 이상, 5.3 중량% 이상, 5.59 중량% 이상, 5.7 중량% 이상, 6 중량% 이상, 7 중량% 이상, 8 중량% 이상, 9 중량% 이상, 10 중량% 이상, 11 중량% 이상, 12 중량% 이상, 13 중량% 이상 또는 14 중량% 이상일 수 있다. 다른 측면에서 상기 GCG는 상기 추출물 총 중량을 기준으로 15 중량% 이하, 14 중량% 이하, 13 중량% 이하, 12 중량% 이하, 11 중량% 이하, 10 중량% 이하, 9 중량% 이하, 8 중량% 이하, 7 중량% 이하, 6 중량% 이하, 5.7 중량% 이하, 5.59 중량% 이하, 5.3 중량% 이하 또는 5 중량% 이하일 수 있다. 상기 GCG가 상기 함량 범위 내로 포함되는 경우에 우수한 피부 미백 활성을 나타낼 수 있다.In one aspect, the GCG is 4% by weight or more, 5% by weight or more, 5.3% by weight or more, 5.59% by weight or more, 5.7% by weight or more, 6% by weight or more, 7% by weight or more, 8% by weight based on the total weight of the extract % Or more, 9% or more, 10% or more, 11% or more, 12% or more, 13% or more, or 14% or more. In another aspect, the GCG is 15 wt% or less, 14 wt% or less, 13 wt% or less, 12 wt% or less, 11 wt% or less, 10 wt% or less, 9 wt% or less, 8 wt% based on the total weight of the extract %, 7% or less, 6% or less, 5.7% or less, 5.59% or less, 5.3% or less, or 5% or less. When the GCG is included within the content range, it may exhibit excellent skin whitening activity.
일 측면에서, 상기 EGCG는 상기 추출물 총 중량을 기준으로 4 중량% 이상, 5 중량% 이상, 5.2 중량% 이상, 5.27 중량% 이상, 5.5 중량% 이상, 6 중량% 이상, 7중량% 이상, 8중량% 이상, 9 중량% 이상, 10 중량% 이상, 11 중량% 이상, 12 중량% 이상, 13 중량% 이상 또는 14 중량% 이상일 수 있다. 다른 측면에서, 상기 EGCG는 상기 추출물 총 중량을 기준으로 15 중량% 이하, 14 중량% 이하, 13 중량% 이하, 12 중량% 이하, 11중량% 이하, 10중량% 이하, 9중량% 이하, 8 중량% 이하, 7 중량% 이하, 6 중량% 이하, 5.5 중량% 이하, 5.27 중량% 이하, 5.2 중량% 이하 또는 5 중량% 이하일 수 있다.In one aspect, the EGCG is based on the total weight of the extract 4 wt% or more, 5 wt% or more, 5.2 wt% or more, 5.27 wt% or more, 5.5 wt% or more, 6 wt% or more, 7 wt% or more, 8 It may be at least 10% by weight, at least 9% by weight, at least 10% by weight, at least 11% by weight, at least 12% by weight, at least 13% by weight, or at least 14% by weight. In another aspect, the EGCG is 15% by weight or less, 14% by weight or less, 13% by weight or less, 12% by weight or less, 11% by weight or less, 10% by weight or less, 9% by weight or less, 8 based on the total weight of the extract It may be up to 7% by weight, up to 6%, up to 5.5%, up to 5.27%, up to 5.2% or up to 5%.
다른 구현 예로서, 상기 추출물 내의 GCG 및 EGCG의 총 함량은 상기 조성물 또는 추출물 총 중량을 기준으로 30 중량% 이하일 수 있다. 일 측면에서 상기 GCG 및 EGCG의 총 함량은 상기 추출물 총 중량을 기준으로 30 중량% 이하, 25 중량% 이하, 20 중량% 이하, 18 중량% 이하, 16 중량% 이하, 15 중량% 이하, 14 중량% 이하, 12 중량% 이하, 10.5 중량% 이하, 10 중량% 이하 또는 9 중량% 이하일 수 있다. 다른 측면에서 상기 GCG 및 EGCG의 총 함량은 상기 추출물 총 중량을 기준으로 8 중량% 이상, 9 중량% 이상, 10 중량% 이상, 10.5 중량% 이상, 12 중량% 이상, 13 중량% 이상, 14 중량% 이상, 16 중량% 이상, 18 중량% 이상, 20 중량% 이상 또는 25 중량% 이상일 수 있다.As another embodiment, the total content of GCG and EGCG in the extract may be 30% by weight or less based on the total weight of the composition or extract. In one aspect, the total content of the GCG and EGCG is 30% by weight or less, 25% by weight or less, 20% by weight or less, 18% by weight or less, 16% by weight or less, 15% by weight or less, 14% by weight based on the total weight of the extract % Or less, 12% or less, 10.5% or less, 10% or less, or 9% or less. In another aspect, the total content of the GCG and EGCG is 8% by weight, 9% by weight, 10% by weight, 10.5% by weight, 12% by weight, 13% by weight, 14% by weight based on the total weight of the extract % Or more, 16% or more, 18% or more, 20% or more, or 25% or more.
다른 구현 예로서, 상기 추출물 내의 에피 카테킨 함량은 상기 추출물 총 중량을 기준으로 20 중량% 이하일 수 있다. 일 측면에서 상기 에피 카테킨 함량은 상기 추출물 총 중량을 기준으로 20 중량% 이하, 18 중량% 이하, 16 중량% 이하, 15 중량% 이하, 14 중량% 이하, 12 중량% 이하, 11 중량% 이하 또는 10 중량% 이하일 수 있다. 다른 측면에서 상기 에피 카테킨 함량은 상기 추출물 총 중량을 기준으로 9 중량% 이상, 10 중량% 이상, 11 중량% 이상, 12 중량% 이상, 13 중량% 이상, 14 중량% 이상, 16 중량% 이상, 또는 18 중량% 이상일 수 있다.As another embodiment, the epicatechin content in the extract may be 20% by weight or less based on the total weight of the extract. In one aspect, the epicatechin content is 20 wt% or less, 18 wt% or less, 16 wt% or less, 15 wt% or less, 14 wt% or less, 12 wt% or less, 11 wt% or less based on the total weight of the extract, or It may be 10% by weight or less. In another aspect, the epicatechin content is at least 9% by weight, at least 10% by weight, at least 11% by weight, at least 12% by weight, at least 13% by weight, at least 14% by weight, at least 16% by weight based on the total weight of the extract, Or 18% or more.
다른 구현 예로서, 상기 추출물 내의 8종 카테킨의 총 함량, 즉, 상기 추출물 내의 EGCG, (-)-에피갈로카테킨((-)-epigallocatechin, EGC), (-)에피카테킨((-)epicattechin, EC), 에피카테킨 3-O-갈레이트(epicatechin 3-O-gallate, ECG), GCG, 갈로카테킨(gallocatechin, GC), 카테킨(catechin, C) 및 카테킨 갈레이트(catechin gallate, CG)의 총 함량은 상기 추출물 총 중량을 기준으로 19 내지 30 중량%일 수 있다. 일 측면에서 상기 8종 카테킨의 총 함량은 상기 추출물 총 중량을 기준으로 19 중량% 이상, 21 중량% 이상, 23 중량% 이상, 24 중량% 이상, 24.5 중량% 이상, 25 중량% 이상, 26 중량% 이상, 27 중량% 이상, 28 중량% 이상 또는 29 중량% 이상일 수 있다. 다른 측면에서 상기 8종 카테킨의 총 함량은 상기 추출물 총 중량을 기준으로 30 중량% 이하, 29 중량% 이하, 28 중량% 이하, 27 중량% 이하, 26 중량% 이하, 25 중량% 이하, 24.5 중량% 이하, 24 중량% 이하, 23 중량% 이하 또는 21 중량% 이하일 수 있다.As another embodiment, the total content of 8 catechins in the extract, i.e., EGCG, (-)-epigallocatechin (EGC), (-) epicatechin ((-)epicattechin, in the extract) EC), the total content of epicatechin 3-O-gallate (ECG), GCG, gallocatechin (GC), catechin (C) and catechin gallate (CG) Silver may be 19 to 30% by weight based on the total weight of the extract. In one aspect, the total content of the eight catechins is 19% by weight or more, 21% by weight or more, 23% by weight or more, 24% by weight or more, 24.5% by weight or more, 25% by weight or more, 26% by weight based on the total weight of the extract % Or more, 27% or more, 28% or more, or 29% or more. In another aspect, the total content of the eight catechins is 30% by weight or less, 29% by weight or less, 28% by weight or less, 27% by weight or less, 26% by weight or less, 25% by weight or less, 24.5% by weight based on the total weight of the extract % Or less, 24% or less, 23% or less, or 21% or less.
또 다른 구현 예로서, 상기 추출물 내의 GCG:EGCG의 중량비는 1:0.5 내지 2일 수 있다. 일 측면에서 상기 추출물 내의 GCG:EGCG의 중량비는 1:0.5, 1:0.8, 1:1, 1:1.2, 1:1.5, 1:1.8 또는 1:2일 수 있다.As another embodiment, the weight ratio of GCG:EGCG in the extract may be 1:0.5 to 2. In one aspect, the weight ratio of GCG:EGCG in the extract may be 1:0.5, 1:0.8, 1:1, 1:1.2, 1:1.5, 1:1.8 or 1:2.
또 다른 구현 예로서, 상기 추출물은 물 및 C1 내지 C4의 알코올 중 어느 하나 이상에 의해 1회 이상 추출한 추출물일 수 있다. 일 측면에서 상기 알코올은 에탄올일 수 있다. 다른 측면에서 상기 알코올은 20% 이상, 30% 이상, 40% 이상, 50% 이상, 60% 이상 또는 70% 이상의 에탄올일 수 있다. 또 다른 측면에서 상기 알코올은 70% 이하, 60% 이하, 50% 이하, 40% 이하 또는 30% 이하의 에탄올일 수 있다. As another embodiment, the extract may be an extract extracted one or more times by any one or more of water and C1 to C4 alcohol. In one aspect, the alcohol may be ethanol. In another aspect, the alcohol may be 20% or more, 30% or more, 40% or more, 50% or more, 60% or more, or 70% or more ethanol. In another aspect, the alcohol may be 70% or less, 60% or less, 50% or less, 40% or less, or 30% or less ethanol.
또 다른 구현 예로서, 상기 피부 미백용 조성물 내 상기 녹차 추출물의 함량은 상기 조성물 전체 중량 대비 1중량% 내지 100중량%일 수 있다. 일 측면에서, 상기 조성물 내 상기 추출물의 함량은 1중량%이상, 10중량%이상, 20중량%이상, 30중량%이상, 40중량%이상, 50중량%이상, 60중량%이상, 70중량%이상, 80중량%이상 또는 90중량%이상일 수 있다. 다른 측면에서, 상기 조성물 내 상기 추출물의 함량은 100중량%이하, 90중량%이하, 80중량%이하, 70중량%이하, 60중량%이하, 50중량%이하, 40중량%이하, 30중량%이하 또는 20중량%이하일 수 있다.As another embodiment, the content of the green tea extract in the composition for skin whitening may be 1% to 100% by weight based on the total weight of the composition. In one aspect, the content of the extract in the composition is 1% by weight, 10% by weight, 20% by weight, 30% by weight, 40% by weight, 50% by weight, 60% by weight, 70% by weight Or more, 80% by weight or more, or 90% by weight or more. In another aspect, the content of the extract in the composition is 100% or less, 90% or less, 80% or less, 70% or less, 60% or less, 50% or less, 40% or less, 30% or less It may be less than or equal to 20% by weight.
다른 구현 예로서, 상기 유효성분의 투여량은 5mg/kg/일 내지 1000mg/kg/일일 수 있다. 일 측면에서, 상기 투여량은 5mg/kg/일 이상, 100mg/kg/일 이상, 200mg/kg/일 이상, 300mg/kg/일 이상, 400mg/kg/일 이상, 500mg/kg/일 이상, 600mg/kg/일 이상, 700mg/kg/일 이상, 800mg/kg/일 이상, 또는 900mg/kg/일 이상일 수 있다. 다른 측면에서, 상기 투여량은 1000mg/kg/일 이하, 900mg/kg/일 이하, 800mg/kg/일 이하, 700mg/kg/일 이하, 600mg/kg/일 이하, 500mg/kg/일 이하, 400mg/kg/일 이하, 300mg/kg/일 이하, 200mg/kg/일 이하, 100mg/kg/일 이하, 50mg/kg/일 이하 또는 10mg/kg/일 이하일 수 있다.As another embodiment, the dosage of the active ingredient may be 5mg/kg/day to 1000mg/kg/day. In one aspect, the dosage is 5 mg/kg/day or more, 100 mg/kg/day or more, 200 mg/kg/day or more, 300 mg/kg/day or more, 400 mg/kg/day or more, 500 mg/kg/day or more, It may be 600 mg/kg/day or more, 700 mg/kg/day or more, 800 mg/kg/day or more, or 900 mg/kg/day or more. In another aspect, the dosage is 1000 mg/kg/day or less, 900 mg/kg/day or less, 800 mg/kg/day or less, 700 mg/kg/day or less, 600 mg/kg/day or less, 500 mg/kg/day or less, It may be 400 mg/kg/day or less, 300 mg/kg/day or less, 200 mg/kg/day or less, 100 mg/kg/day or less, 50 mg/kg/day or less, or 10 mg/kg/day or less.
일 구현 예로서, 상기 추출물은 멜라닌 생성을 억제할 수 있으며, 다른 구현 예로서, 상기 추출물은 티로시나아제(tyrosinase)의 활성을 억제할 수 있다. 즉, 상기 추출물 또는 조성물은 멜라닌 생성을 억제하거나 티로시나아제의 활성을 억제하여 피부 미백 효과를 나타낼 수 있다.As an embodiment, the extract may inhibit melanin production, and as another embodiment, the extract may inhibit tyrosinase activity. That is, the extract or composition may exhibit skin whitening effect by inhibiting melanin production or inhibiting tyrosinase activity.
일 구현 예로서, 상기 조성물은 식품, 약학 또는 화장료 조성물일 수 있다.As an example embodiment, the composition may be a food, pharmaceutical or cosmetic composition.
상기 식품 조성물의 제형은 특별히 한정되지 않으나, 예를 들어, 정제, 과립제, 환제, 분말제, 드링크제와 같은 액제, 캐러멜, 겔, 바, 티백 등으로 제형화될 수 있다. 각 제형의 식품 조성물은 유효 성분 이외에 해당 분야에서 통상적으로 사용되는 성분들을 제형 또는 사용 목적에 따라 당업계의 통상의 기술자가 어려움 없이 적의 선정하여 배합할 수 있으며, 다른 원료와 동시에 적용할 경우 상승 효과가 일어날 수 있다. 또한 상기 식품은 건강기능식품일 수도 있다.The formulation of the food composition is not particularly limited, for example, tablets, granules, pills, powders, liquids such as drinks, caramels, gels, bars, tea bags, etc. may be formulated. The food composition of each formulation can be formulated by appropriately selecting the ingredients commonly used in the field other than the active ingredients according to the formulation or purpose of use without difficulty, and synergistic effect when applied simultaneously with other ingredients. Can happen. In addition, the food may be a health functional food.
상기 조성물은 단순 섭취, 음용, 주사 투여, 스프레이 투여 또는 스퀴즈 투여 등 다양한 방법으로 투여될 수 있다. The composition may be administered by various methods, such as simple ingestion, drinking, injection administration, spray administration, or squeeze administration.
본 발명의 일 측면에 따른 식품 조성물에 있어서, 상기 유효 성분의 투여량 결정은 당업계의 통상의 기술자의 수준 내에 있으며, 투여하고자 하는 대상의 연령, 건강 상태, 합병증 등 다양한 요인에 따라 달라질 수 있다.In the food composition according to one aspect of the present invention, the dosage of the active ingredient is within the level of those skilled in the art, and may vary depending on various factors such as age, health condition, and complications of the subject to be administered. .
본 발명의 일 측면에 따른 식품 조성물은, 예를 들어, 츄잉껌, 캐러멜 제품, 캔디류, 빙과류, 과자류 등의 각종 식품류, 청량 음료, 미네랄 워터, 알코올 음료 등의 음료 제품, 비타민이나 미네랄 등을 포함한 건강기능식품 제품일 수 있다.Food composition according to an aspect of the present invention, for example, chewing gum, caramel products, candy, ice cream, confectionery, various foods, soft drinks, mineral water, alcoholic beverages, beverage products, vitamins, minerals, etc. It may be a functional food product.
상기 외에, 본 발명의 일 측면에 따른 식품 조성물은 여러 가지 영양제, 비타민, 광물(전해질), 합성 풍미제 및 천연 풍미제 등의 풍미제, 착색제 및 증진제(치즈, 초콜릿 등), 펙트산 및 그의 염, 알긴산 및 그의 염, 유기산, 보호성 콜로이드 증점제, pH 조절제, 안정화제, 방부제, 글리세린, 알콜, 탄산 음료에 사용되는 탄산화제 등을 포함할 수 있다. 그 밖에 본 발명의 일측면에 따른 식품 조성물들은 천연 과일 쥬스 및 과일 쥬스 음료 및 야채 음료의 제조를 위한 과육을 포함할 수 있다. 이러한 성분은 독립적으로 또는 조합하여 사용할 수 있다. 이러한 첨가제의 비율은 그렇게 중요하진 않으나 본 발명의 일측면에 따른 조성물 100 중량부 당 0 내지 약 60 중량부의 범위에서 포함되는 것이 일반적이다.In addition to the above, the food composition according to one aspect of the present invention includes various nutrients, vitamins, minerals (electrolytes), flavoring agents such as synthetic flavoring agents and natural flavoring agents, colorants and enhancers (cheese, chocolate, etc.), pectic acid and the like. Salts, alginic acids and salts thereof, organic acids, protective colloidal thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonic acid used in carbonated beverages, and the like. In addition, food compositions according to an aspect of the present invention may include natural fruit juice and fruit juice for the production of fruit juice drinks and vegetable drinks. These ingredients can be used independently or in combination. The proportion of these additives is not so important, but is generally included in the range of 0 to about 60 parts by weight per 100 parts by weight of the composition according to one aspect of the present invention.
본 발명의 일 측면에 따른 상기 약학 조성물은 경구, 비경구, 직장, 국소, 경피, 정맥 내, 근육 내, 복강 내, 피하 등으로 투여될 수 있다. 경구 투여를 위한 제형은 정제(錠劑), 환제(丸劑), 연질 및 경질 캅셀제, 과립제(顆粒劑), 산제, 세립제, 액제, 유탁제(乳濁濟) 또는 펠렛제일 수 있으나, 이에 제한되는 것은 아니다. 비경구 투여를 위한 제형은 용액제, 현탁제, 유액제, 겔, 주사제, 점적제, 좌제(坐劑), 패취 또는 분무제일 수 있으나, 이에 제한되는 것은 아니다. 상기 제형은 당해 분야의 통상적인 방법에 따라 용이하게 제조될 수 있으며, 계면 활성제, 부형제, 수화제, 유화 촉진제, 현탁제, 삼투압 조절을 위한 염 또는 완충제, 착색제, 향신료, 안정화제, 방부제, 보존제 또는 기타 상용하는 보조제를 추가로 포함할 수 있다.The pharmaceutical composition according to an aspect of the present invention may be administered by oral, parenteral, rectal, topical, transdermal, intravenous, intramuscular, intraperitoneal, subcutaneous, and the like. Formulations for oral administration may be tablets, pills, soft and hard capsules, granules, powders, granules, liquids, emulsions or pellets, but are not limited thereto. It is not. Formulations for parenteral administration may be solutions, suspensions, emulsions, gels, injections, drops, suppositories, patches or sprays, but are not limited thereto. The formulations can be readily prepared according to conventional methods in the art, surfactants, excipients, hydrating agents, emulsifying accelerators, suspending agents, salts or buffers for controlling osmotic pressure, colorants, spices, stabilizers, preservatives, preservatives or Other commercial adjuvants may be further included.
본 발명의 일 측면에 따른 상기 조성물은 약학적으로 허용가능한 염을 포함할 수도 있는데, 상기 염은 (1) 염산, 브롬화수소산, 황산, 질산, 인산 등과 같은 무기산으로 형성되거나; 또는 아세트산, 프로파이온산, 헥사노산, 시클로펜테인프로피온산, 글라이콜산, 피루브산, 락트산, 말론산, 숙신산, 말산, 말레산, 푸마르산, 타르타르산, 시트르산, 벤조산, 3-(4-히드록시벤조일) 벤조산, 신남산, 만델산, 메테인설폰산, 에테인설폰산, 1,2-에테인-디설폰산, 2-히드록시에테인설폰산, 벤젠설폰산, 4-클로로벤젠설폰산, 2-나프탈렌설폰산, 4-톨루엔설폰산, 캄퍼설폰산, 4-메틸바이시클로 [2,2,2]-oct-2-엔-1-카르복실산, 글루코헵톤산, 3-페닐프로파이온산, 트리메틸아세트산, tert-부틸아세트산, 라우릴 황산, 글루콘산, 글루탐산, 히드록시나프토산, 살리실산, 스테아르산, 뮤콘산과 같은 유기산으로 형성되는 산 부가염(acid addition salt); 또는 (2) 모 화합물에 존재하는 산성 프로톤이 치환될 때 형성되는 염을 포함할 수 있다.The composition according to an aspect of the present invention may include a pharmaceutically acceptable salt, which salt is formed of an inorganic acid such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, etc.; Or acetic acid, propionic acid, hexanoic acid, cyclopentanepropionic acid, glycolic acid, pyruvic acid, lactic acid, malonic acid, succinic acid, malic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, 3-(4-hydroxybenzoyl) Benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, 1,2-ethane-disulfonic acid, 2-hydroxyethanesulfonic acid, benzenesulfonic acid, 4-chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid, 4-toluenesulfonic acid, camphorsulfonic acid, 4-methylbicyclo [2,2,2]-oct-2-ene-1-carboxylic acid, glucoheptonic acid, 3-phenylpropionic acid, trimethylacetic acid, tert Acid addition salts formed from organic acids such as butylacetic acid, lauryl sulfate, gluconic acid, glutamic acid, hydroxynaphthoic acid, salicylic acid, stearic acid, and muconic acid; Or (2) a salt formed when the acidic proton present in the parent compound is substituted.
본 발명의 일측면에 따른 상기 약학 조성물의 적용량 또는 투여량은 투여 받을 대상의 연령, 성별, 체중, 병리 상태 및 그 심각도, 투여 경로 또는 처방자의 판단에 따라 달라질 것이다. 이러한 인자에 기초한 유효성분 투여량 결정은 당업계 통상의 기술자의 수준 내에 있다.The applied amount or dosage of the pharmaceutical composition according to an aspect of the present invention will vary depending on the age, gender, weight, pathology and severity of the subject to be administered, the route of administration or the judgment of the prescriber. The determination of the active ingredient dosage based on these factors is within the level of those skilled in the art.
본 발명의 일 측면에 따른 상기 화장료 조성물은 화장품학 또는 피부과학적으로 허용 가능한 매질 또는 기제를 함유할 수 있다. 이는 국소적용에 적합한 모든 제형으로, 예를 들면, 용액, 겔, 고체, 반죽 무수 생성물, 수상에 유상을 분산시켜 얻은 에멀젼, 현탁액, 마이크로에멀젼, 마이크로캡슐, 미세과립구 또는, 이온형(리포좀) 및 비이온형의 소낭 분산제의 형태로, 또는 크림, 스킨, 로션, 파우더, 연고, 스프레이 또는 콘실 스틱의 형태로 제공될 수 있다. 이들 조성물은 당해 분야의 통상적인 방법에 따라 제조될 수 있다. 상기 화장료 조성물은 또한 포말(foam)의 형태로 압축된 추진제를 더 함유한 에어로졸 조성물의 형태로도 사용될 수 있다.The cosmetic composition according to an aspect of the present invention may contain a cosmetic or dermatologically acceptable medium or base. These are all formulations suitable for topical application, e.g. solutions, gels, solids, dough anhydrous products, emulsions, suspensions, microemulsions, microcapsules, microgranules, or ionic (liposomes) obtained by dispersing an oil phase in an aqueous phase, and It may be provided in the form of a non-ionic vesicle dispersant, or in the form of a cream, skin, lotion, powder, ointment, spray or conceal stick. These compositions can be prepared according to conventional methods in the art. The cosmetic composition may also be used in the form of an aerosol composition further containing a propellant compressed in the form of a foam.
상기 화장료 조성물은 그 제형에 있어서 특별히 한정되는 바가 없으며, 목적하는 바에 따라 적절히 선택될 수 있다. 예를 들어, 스킨로션, 스킨소프너, 스킨토너, 로션, 밀크로션, 모이스쳐 로션, 영양로션, 맛사지크림, 영양크림, 모이스처크림, 핸드크림, 파운데이션, 에센스, 영양에센스, 팩, 비누, 클렌징폼, 클렌징로션, 클렌징크림, 클렌징워터, 파우더, 바디로션, 바디크림, 바디오일, 바디클렌저 및 바디에센스 등의 제형으로 제조될 수 있다.The cosmetic composition is not particularly limited in its formulation, and may be appropriately selected according to the purpose. For example, skin lotion, skin softener, skin toner, lotion, milk lotion, moisture lotion, nutrition lotion, massage cream, nutrition cream, moisture cream, hand cream, foundation, essence, nutrition essence, pack, soap, cleansing foam, Cleansing lotions, cleansing creams, cleansing water, powders, body lotions, body creams, body oils, body cleansers, and body essences can be formulated into formulations.
본 발명의 제형이 페이스트, 크림 또는 겔인 경우에는 담체 성분으로서 동물섬유, 식물섬유, 왁스, 파라핀, 전분, 트라칸트, 셀룰로오스 유도체, 폴리에틸렌 글리콜, 실리콘, 벤토나이트, 실리카, 탈크 또는 산화아연 등이 이용될 수 있다.When the formulation of the present invention is a paste, cream or gel, animal fibers, plant fibers, wax, paraffin, starch, tracant, cellulose derivatives, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide, etc. may be used as a carrier component. Can.
본 발명의 제형이 파우더 또는 스프레이인 경우에는 담체 성분으로서 락토스, 탈크, 실리카, 알루미늄히드록시드, 칼슘 실리케이트 또는 폴리아미드 파우더가 이용될 수 있고, 특히 스프레이인 경우에는 추가적으로 클로로플루오로히드로카본, 프로판/부탄 또는 디메틸 에테르와 같은 추진체를 포함할 수 있다.When the formulation of the present invention is a powder or spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as a carrier component, and in particular, in the case of a spray, additionally chlorofluorohydrocarbon, propane /Propellant such as butane or dimethyl ether.
본 발명의 제형이 용액 또는 유탁액의 경우에는 담체 성분으로서 용매, 용매화제 또는 유탁화제가 이용되고, 예컨대 물, 에탄올, 이소프로판올, 에틸 카보네이트, 에틸 아세테이트, 벤질 알코올, 벤질 벤조에이트, 프로필렌 글리콜, 1,3-부틸글리콜 오일, 글리세롤 지방족 에스테르, 폴리에틸렌 글리콜 또는 소르비탄의 지방산 에스테르가 있다.When the formulation of the present invention is a solution or emulsion, a solvent, solvating agent or emulsifying agent is used as a carrier component, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1 Fatty acid esters of ,3-butyl glycol oil, glycerol aliphatic esters, polyethylene glycol or sorbitan.
본 발명의 제형이 현탁액인 경우에는 담체 성분으로서 물, 에탄올 또는 프로필렌 글리콜과 같은 액상 희석제, 에톡실화 이소스테아릴 알코올, 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, liquid diluents such as water, ethanol or propylene glycol as carrier components, ethoxylated isostearyl alcohol, suspensions such as polyoxyethylene sorbitol esters and polyoxyethylene sorbitan esters, microcrystalline Cellulose, aluminum metahydroxide, bentonite, agar or trakant, etc. can be used.
본 발명의 제형이 계면-활성제 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, 식물성 유, 리놀린 유도체 또는 에톡실화 글리세롤 지방산 에스테르 등이 이용될 수 있다.When the formulation of the present invention is a surfactant-containing cleansing, aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivatives, methyltaurate, sarcosinate, fatty acid amide as a carrier component Ether sulfates, alkylamidobetaines, aliphatic alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, linoline derivatives or ethoxylated glycerol fatty acid esters and the like can be used.
상기 화장료 조성물에는 상기 녹차 추출물 이외에 기능성 첨가물 및 일반적인 화장료 조성물에 포함되는 성분이 추가로 포함될 수 있다. 상기 기능성 첨가물로는 수용성 비타민, 유용성 비타민, 고분자 펩티드, 고분자 다당, 스핑고 지질 및 해초 엑기스로 이루어지는 군에서 선택된 성분을 포함할 수 있다.In addition to the green tea extract, the cosmetic composition may further include functional additives and components included in the general cosmetic composition. The functional additive may include a component selected from the group consisting of water-soluble vitamins, oil-soluble vitamins, polymer peptides, polymer polysaccharides, sphingo lipids, and seaweed extract.
상기 조성물에는 또한, 상기 기능성 첨가물과 더불어 필요에 따라 일반적인 화장료 조성물에 포함되는 성분을 배합해도 된다. 이외에 포함되는 배합 성분으로서는 유지 성분, 보습제, 에몰리엔트제, 계면 활성제, 유기 및 무기 안료, 유기 분체, 자외선 흡수제, 방부제, 살균제, 산화 방지제, 식물 추출물, pH 조절제, 알코올, 색소, 향료, 혈행 촉진제, 냉감제, 제한(制汗)제, 정제수 등을 들 수 있다.In addition to the above-mentioned functional additives, the components contained in the general cosmetic composition may be blended with the composition. Other ingredients included include fats and oils, moisturizers, emollients, surfactants, organic and inorganic pigments, organic powders, UV absorbers, preservatives, fungicides, antioxidants, plant extracts, pH regulators, alcohols, pigments, flavors, blood circulation Accelerators, cooling agents, limiting agents, purified water, and the like.
이하, 실시예, 실험예, 및 제형예를 들어 본 명세서의 구성 및 효과를 보다 구체적으로 설명한다. 그러나 이들 예는 본 명세서에 대한 이해를 돕기 위해 예시의 목적으로만 제공된 것일 뿐 본 명세서의 범주 및 범위가 하기 예에 의해 제한되는 것은 아니다.Hereinafter, examples, experimental examples, and formulation examples, the composition and effects of the present specification will be described in more detail. However, these examples are provided for purposes of illustration only to aid understanding of the present specification, and the scope and scope of the present specification are not limited by the following examples.
[실시예 1] 일반 고시형 녹차 추출물과 고온처리 녹차 추출물의 제조[Example 1] Preparation of general notice green tea extract and high temperature treated green tea extract
100g의 녹차(Camellia sinensis, 제주 오설록 농장)에 50% 에탄올 1000ml 를 가하여 60℃에서 1시간 환류 교반하였다. 상기 시료의 온도를 실온으로 낮추고 여과하여 얻은 용액을 감압증류하여 진한갈색분말의 일반 고시형 녹차 추출물(GT-LE-35CAT, 시료 1) 23g을 수득하였다(수율 23%).To 100 g of green tea (Camellia sinensis, Jeju Osulloc Farm), 1000 ml of 50% ethanol was added and stirred under reflux at 60° C. for 1 hour. The solution obtained by lowering the temperature of the sample to room temperature and filtering was distilled under reduced pressure to obtain 23 g of a general notice green tea extract (GT-LE-35CAT, sample 1) of a dark brown powder (yield 23%).
한편, 고온처리 녹차 추출물을 제조하기 위하여, 100g의 녹차(Camellia sinensis, 제주 오설록 농장)에 50% 에탄올 1000ml를 가하여 60℃에서 1시간 환류 교반하였다. 이것을 농축 후 스팀 1.5 kgf/cm2에서 1~7시간 교반시켰다. 이후 온도를 실온으로 낮추고 불용성물질을 여과하고 감압농축하여 고온처리 녹차 추출물 10g을 수득하였다. 이 때, 상기 교반 시간대(1-7시간) 별로 얻은 상기 고온처리 녹차 추출물에 대해 하기 표 1과 같은 장치를 이용하여 8종 카테킨의 함량 변화를 측정하였고, 5시간 째에 열에 의한 EGCG의 GCG로의 전환이 가장 많으면서, 8종 카테킨의 양도 더 이상 줄어들지 않음을 확인하였다. 이때 수득한 고온처리 녹차 추출물 10g(High temperature processed green tea extract, HTP-GTE)을 시료 2로 하였다.Meanwhile, 1000 ml of 50% ethanol was added to 100 g of green tea (Camellia sinensis, Jeju Osulloc Farm) to prepare a high-temperature-treated green tea extract, followed by stirring at reflux at 60° C. for 1 hour. After concentration, it was stirred for 1 to 7 hours at 1.5 kgf/cm 2 of steam. Thereafter, the temperature was lowered to room temperature, the insoluble material was filtered, and concentrated under reduced pressure to obtain 10 g of a hot-treated green tea extract. At this time, the change in the content of 8 catechins was measured using the apparatus shown in Table 1 below for the high-temperature treated green tea extract obtained for each stirring time period (1-7 hours), and heat conversion of EGCG to GCG by heat at 5 hours. It was confirmed that the most conversion, the amount of 8 kinds of catechins did not decrease any more. At this time, the obtained high temperature processed green tea extract 10g (High temperature processed green tea extract, HTP-GTE) was used as Sample 2.
수득된 상기 두 종류의 추출물의 조성을 분석한 조건과 결과는 각각 하기 표 1(시료 1 및 2의 조성 분석 조건), 표 2(시료 1의 조성 분석 결과) 및 표 3(시료 2의 조성 분석 결과)와 같다. 또한, 상기 두 종류의 추출물에 대한 크로마토그램은 도 1(시료 1) 및 도 2(시료 2)와 같다. 즉, 시료 2의 경우 기존의 녹차 추출물과는 조성이 다르다는 것을 확인하였다. 구체적으로, 시료 2의 경우 시료 1에 비해 현저하게 낮은 EGCG(5.27중량%), EGC(3.53중량%) 및 총 카테킨(24.41중량%)을 함유하는 반면, 시료 1에서는 발견되지 않은 성분인 4종의 에피 카테킨 에피머가 추가로 생성되는 것을 확인할 수 있었다.The conditions and results of analyzing the composition of the obtained two types of extracts are shown in Table 1 (composition analysis conditions of
42min A(80) : B(20)
44min A(5) : B(95)
49min A(90) : B(10)0 min A(90): B(10)30min A(85): B(15)
42min A(80): B(20)
44min A(5): B(95)
49min A(90): B(10)
(상기 표 2 및 표 3에서, GC는 갈로카테킨, EGC는 에피갈로카테킨, C는 카테킨, EC는 (-)에피카테킨, GCG는 갈로카테킨 갈레이트, EGCG는 에피갈로카테킨 갈레이트, CG는 카테킨 갈레이트, ECG는 에피카테킨 3-O-갈레이트)(In Tables 2 and 3, GC is gallocatechin, EGC is epigallocatechin, C is catechin, EC is (-) epicatechin, GCG is gallocatechin gallate, EGCG is epigallocatechin gallate, CG is Catechin gallate, ECG is epicatechin 3-O-gallate)
(상기 표 2 및 표 3의 단위는 모두 녹차 추출물(시료 1 및 2) 전체 중량 대비 해당 물질의 중량%)(All of the units in Tables 2 and 3 above are green tea extracts (
[실험예 1] 고온처리 녹차 추출물의 세포 독성 확인 실험[Experimental Example 1] Cytotoxicity test of high temperature treated green tea extract
Lonza에서 구한 Melanocyte 세포주(피부 세포종)를 96웰 플레이트 (well plate, FALCON)에 웰 당 1X105씩 시딩하고(seeding), 24시간 동안 37℃, 5% CO2 인큐베이터에서 배양 후, 상기 실시예 1의 고온처리 녹차 추출물(시료 2)을 10 ppm 및 100 ppm 으로 처리하여 48시간 동안 추가 배양하였다.Melanocyte cell line (skin cell tumor) obtained from Lonza was seeded in a 96-well plate (well plate, FALCON) at a rate of 1×10 5 per well, cultured in a 5% CO 2 incubator at 37° C. for 24 hours, and then in Example 1 High temperature treatment of green tea extract (Sample 2) was treated with 10 ppm and 100 ppm for further incubation for 48 hours.
이후 배지를 제거하고 cell counting kit-8 (Dojindo)를 이용하여 세포 생존율을 확인하였다. RPMI1640 (Lonza) 100 μl에 cell counting kit-8 (Dojindo)용액 10 μl를 첨가하여 세포에 처리 후 450 nm에서 흡광도를 확인하여 살아있는 세포의 수를 정량화 하였다. 세포 수 또는 세포 생존율(%)는 하기 식에 의해 계산되었다.After that, the medium was removed and cell viability was checked using a cell counting kit-8 (Dojindo). After adding 10 μl of cell counting kit-8 (Dojindo) solution to 100 μl of RPMI1640 (Lonza), the cells were treated and the absorbance at 450 nm was checked to quantify the number of live cells. Cell number or cell viability (%) was calculated by the following formula.
세포 생존율 (%) = (시료 처리군 흡광도 - 반응시약만의 흡광도) / (무처리군 흡광도 - 반응시약만의 흡광도) X 100Cell viability (%) = (Absorbance of the sample treated group-Absorbance of only the reaction reagent) / (Absorbance of the untreated group-Absorbance of the reaction reagent only)
그 결과는 도 3과 같으며, 10ppm 및 100ppm의 고온처리 녹차 추출물(시료 2) 모두에서 세포 독성이 나타나지 않음을 확인하였다.The results are as shown in FIG. 3, and it was confirmed that no cytotoxicity was observed in both 10ppm and 100ppm high temperature treated green tea extract (Sample 2).
[실험예 2] 고온처리 녹차 추출물의 멜라닌 생성 억제 평가[Experimental Example 2] Evaluation of melanin production inhibition of high-temperature-treated green tea extract
멜라닌 생성이 과도한 인간 흑색종 세포인 MNT-1 세포(human melanoma cell line, Lonza, SWISS)를 6웰 plate에 1x106 cell/well로 시딩(seeding)하고 37℃, 5% CO2 조건 하에서 24동안 배양한 후, 상기 실시예 1의 시료 1과 시료 2를 각각 10ppm 및 100ppm을 처리하여 48시간 동안 37℃, 5% CO2 인큐베이터에서 배양하였다. 48시간 동안 배양된 상기 세포에 파쇄액(Lysis Buffer; 1% NP-40, 50mM Tris-HCL; pH 7.5, 150mM NaCl)을 500μl 처리한 다음, 셀 스크래퍼(Cell scraper)를 사용하여 마이크로튜브(microtube)에 모은 후, 13,000rpm으로 15분 동안 원심분리(Eppendorf, centrifuge 5415R, 독일)하였다. 그 후, 침전물을 분리하여 1N 수산화나트륨 50μl를 처리하여 침전물을 용해시킨 다음, 멜라닌 색소에 특이적인 450nm 파장에 대한 흡광도를 측정하고 하기 계산식에 따라 멜라닌의 양을 전체 단백질 양에 대한 값으로 보정하였다. 이때 대조군(control)으로는 시료를 처리하지 않은 MNT-1 세포를 사용하였으며, 그 결과는 도 4에 나타내었다.MNT-1 cells (human melanoma cell line, Lonza, SWISS), which are human melanoma cells with excessive melanin production, are seeded in a 6-well plate at 1×10 6 cells/well and seeded at 37° C. for 5 hours under 5% CO 2 conditions. After cultivation, the
멜라닌 양 (%) = 흡광도 / 전체 단백질 양Melanin amount (%) = absorbance / total protein amount
도 4의 결과로부터, 일반 녹차 추출물(시료 1)에 비하여 본 발명에 따른 고온처리 녹차 추출물(시료 2)의 멜라닌 생성 억제 효능이 크게 우수함을 확인할 수 있었다.From the results of FIG. 4, it was confirmed that the high-temperature-treated green tea extract according to the present invention (sample 2) had a significantly superior melanin production inhibitory effect compared to the general green tea extract (sample 1).
특히, 고온처리 녹차 추출물을 100ppm 처리한 경우 멜라닌 생성 억제 효과가 현저히 높게 나타났으며, 또한, 고온처리 녹차 추출물 10ppm 처리한 경우, 일반 녹차 추출물을 100ppm 처리한 경우와 유사한 수준의 멜라닌 생성 억제 효과가 나타나, 본 발명에 따른 고온처리 녹차 추출물의 멜라닌 생성 억제 효능이 일반 녹차 추출물에 비하여 10배 가량 뛰어남을 확인할 수 있었다.In particular, when the high-temperature-treated green tea extract was treated at 100 ppm, the melanin production inhibitory effect was significantly higher, and when the high-temperature-treated green tea extract was treated at 10 ppm, the melanin production inhibitory effect was similar to that of the normal green tea extract treated at 100 ppm. It appeared, it was confirmed that the high-temperature-treated green tea extract according to the present invention has an inhibitory effect on melanin production of about 10 times that of a normal green tea extract.
[실험예 3] 고온처리 녹차 추출물의 티로시나아제 활성 억제 평가[Experimental Example 3] Inhibition of tyrosinase activity inhibition of high temperature treated green tea extract
상기 실험예 2에 따른 세포 용해물 10 μg에 DOPA(디하이드록시페닐알라닌; dihydroxyphenylalanine) 10 mM(동량 10 μg)를 첨가하고, 티로시나아제의 작용에 의하여 생성된 도파크롬(dopachrome)의 생성량을 490nm에서의 흡광도를 비교하여 측정하였다. 티로시나아제 활성을 비처리군의 흡광도에 대한 비율로 계산하여 도 5에 나타내었다.10 μg of DOPA (dihydroxyphenylalanine) was added to 10 μg of the cell lysate according to Experimental Example 2, and the amount of dopachrome produced by the action of tyrosinase was 490 nm. The absorbance at was compared and measured. Tyrosinase activity is calculated as a ratio to the absorbance of the untreated group and is shown in FIG. 5.
도 5의 결과로부터, 본 발명에 따른 고온처리 녹차 추출물을 처리한 경우 일반 녹차 추출물을 처리한 경우에 비하여 티로시나아제 활성 억제 효과가 현저히 뛰어남을 확인할 수 있었다.From the results of FIG. 5, it was confirmed that when the high-temperature-treated green tea extract according to the present invention was treated, the tyrosinase activity inhibitory effect was significantly superior to that of the normal green tea extract.
[제형예 1] 연질 캡슐[Formulation Example 1] Soft capsule
상기 실시예 1에 따른 시료 2를 150mg으로 준비하고, 락토오스 440mg, 옥수수 전분 430mg 및 스테아린산 마그네슘 2mg을 혼합하여 연질 캡슐 충진액을 제조하였다. 그리고, 상기와 별도로 젤라틴 66 중량부, 글리세린 24 중량부 및 솔비톨액 10 중량부의 비율로 연질 캡슐 시트를 제조하고 상기 충진액을 충진시켜 연질 캡슐을 제조하였다.Sample 2 according to Example 1 was prepared as 150 mg, and a soft capsule filling solution was prepared by mixing lactose 440 mg, corn starch 430 mg, and magnesium stearate 2 mg. Then, separately from the above, a soft capsule sheet was prepared at a ratio of 66 parts by weight of gelatin, 24 parts by weight of glycerin, and 10 parts by weight of sorbitol solution, and the filling solution was filled to prepare a soft capsule.
[제형예 2] 정제[Formulation Example 2] Tablet
상기 실시예 1에 따른 시료 2를 150mg으로 준비하고, 비타민 E 15mg, 비타민 C 15mg, 갈락토올리고당 250㎎, 유당 60㎎ 및 맥아당 140㎎을 혼합한 뒤 유동층 건조기를 이용하여 과립화한 후 당 에스테르(sugar ester) 8㎎을 첨가하였다. 이 조성물을 통상의 방법으로 타정하여 정제를 제조하였다.Sample 2 according to Example 1 was prepared as 150 mg, mixed with vitamin E 15 mg, vitamin C 15 mg, galactooligosaccharide 250 mg,
[제형예 3] 앰플제[Formulation Example 3] Ampoule
상기 실시예 1에 따른 시료 2를 10mg으로 준비하고, 말토덱스트린를 혼합한 뒤 정제수 15ml을 가하여 앰플 병에 충진한 후 살균하여 액상 앰플제를 제조하였다.Sample 2 according to Example 1 was prepared as 10 mg, maltodextrin was mixed, 15 ml of purified water was added, filled into an ampoule bottle, and sterilized to prepare a liquid ampoule.
[제형예 4] 드링크제[Formulation Example 4] Drink
상기 실시예 1에 따른 시료 2를 80mg 준비하고, 비타민 E 9mg, 비타민 C 9mg, 포도당 10g, 구연산 0.6g, 및 액상 올리고당 25g을 혼합한 후 정제수 400㎖를 가하여 충진하였다. 병에 충진한 후 30℃에서 4~5 초간 살균하여 드링크제를 제조하였다.Sample 2 according to Example 1 was prepared in 80mg, vitamin E 9mg, vitamin C 9mg, glucose 10g, citric acid 0.6g, and liquid oligosaccharide 25g was mixed, and 400ml of purified water was added to fill it. After filling the bottle, sterilized at 30°C for 4 to 5 seconds to prepare a drink.
[제형예 5] 과립제[Formulation Example 5] Granules
상기 실시예 1에 따른 시료 2를 150mg 준비하고, 비타민 E 9mg, 비타민 C 9mg, 무수결정 포도당 250㎎ 및 전분 550㎎을 혼합한 후, 유동층 과립기를 사용하여 과립으로 성형한 뒤 포에 충진하여 과립제를 제조하였다.150 mg of Sample 2 according to Example 1 was prepared, and mixed with vitamin E 9 mg, vitamin C 9 mg, anhydrous crystalline glucose 250 mg, and starch 550 mg, molded into granules using a fluidized bed granulator, and then filled into a bag to form granules. Was prepared.
[제형예 6] 건강 식품[Formulation Example 6] Health Food
상기 실시예 1에 따른 시료 2를 150 mg 준비하고, 비타민 혼합물(비타민 A 아세테이트 70 ㎍, 비타민 E 1.0 ㎎, 비타민 B1 0.13 ㎎, 비타민 B2 0.15 ㎎, 비타민 B6 0.5 ㎎, 비타민 B12 0.2 ㎍, 비타민 C 10 ㎎, 비오틴 10 ㎍, 니코틴산아미드 1.7 ㎎, 엽산 50 ㎍)과 무기질 혼합물(황산제1철 1.75 ㎎, 산화아연 0.82 ㎎, 탄산마그네슘 25.3 ㎎, 제1인산칼륨 15 ㎎, 제2인산칼슘 55 ㎎, 구연산칼륨 90 ㎎, 탄산칼슘 100 ㎎, 염화마그네슘 24.8 ㎎)을 조합하여 건강식품을 제조하였다. Prepare 150 mg of Sample 2 according to Example 1 above, vitamin mixture (70 μg of vitamin A acetate, 1.0 mg of vitamin E, 0.13 mg of vitamin B1, 0.15 mg of vitamin B2, 0.5 mg of vitamin B6, 0.2 mg of vitamin B12,
[제형예 7] 건강 음료[Formulation Example 7] Healthy Drink
상기 실시예 1에 따른 시료 2를 50 mg 준비하고, 구연산 1000 ㎎, 올리고당 100 g, 매실농축액 2 g, 타우린 1 g, 정제수 잔량을 첨가하여 900 mL의 건강음료를 제조하였다. 50 mg of Sample 2 according to Example 1 was prepared, and 1000 mL of citric acid, 100 g of oligosaccharide, 2 g of plum concentrate, 1 g of taurine, and residual amount of purified water were added to prepare 900 mL of healthy beverage.
이상으로 본 명세서의 특정한 부분을 상세히 기술하였는 바, 당업계의 통상의 지식을 가진 자에게 있어서 이러한 구체적인 기술은 단지 바람직한 구현예일 뿐이며, 이에 본 명세서의 범위가 제한되는 것이 아닌 점은 명백하다. 따라서, 본 명세서의 실질적인 범위는 첨부된 청구항과 그의 등가물에 의하여 정의된다고 할 것이다.Since specific parts of the present specification have been described in detail above, it is obvious that for those skilled in the art, this specific technology is only a preferred embodiment, and the scope of the present specification is not limited thereto. Accordingly, the substantial scope of the present specification will be defined by the appended claims and their equivalents.
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KR1020190005248A KR20200088665A (en) | 2019-01-15 | 2019-01-15 | Compoisition for skin-whitening comprising green tea extract which has modified amounts of ingredients |
EP19213633.1A EP3682865A1 (en) | 2019-01-15 | 2019-12-04 | Composition for skin-whitening or preventing or improving skin wrinkles comprising green tea extract which has modified amounts of ingredients |
JP2019226797A JP7453783B2 (en) | 2019-01-15 | 2019-12-16 | Composition for skin whitening or skin wrinkle prevention or improvement containing green tea extract with changed component content |
CN202010035802.1A CN111434333B (en) | 2019-01-15 | 2020-01-14 | Composition for skin whitening or preventing or improving skin wrinkles comprising green tea extract having a modified amount of ingredients |
US16/742,812 US11896640B2 (en) | 2019-01-15 | 2020-01-14 | Composition for skin-whitening or preventing or improving skin wrinkles comprising green tea extract which has modified amounts of ingredients |
US18/520,281 US20240091296A1 (en) | 2019-01-15 | 2023-11-27 | Composition for skin-whitening or preventing or improving skin wrinkles comprising green tea extract which has modified amounts of ingredients |
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KR1020190005248A KR20200088665A (en) | 2019-01-15 | 2019-01-15 | Compoisition for skin-whitening comprising green tea extract which has modified amounts of ingredients |
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KR20170107943A (en) | 2017-09-14 | 2017-09-26 | (주)아모레퍼시픽 | Skin Whitening Cosmetic Composition Containing Extracts of Wikstroemia trichotoma |
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KR20170107943A (en) | 2017-09-14 | 2017-09-26 | (주)아모레퍼시픽 | Skin Whitening Cosmetic Composition Containing Extracts of Wikstroemia trichotoma |
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