KR20230039105A - Skin-lightening Composition Using Xanthatin - Google Patents
Skin-lightening Composition Using Xanthatin Download PDFInfo
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- KR20230039105A KR20230039105A KR1020210122012A KR20210122012A KR20230039105A KR 20230039105 A KR20230039105 A KR 20230039105A KR 1020210122012 A KR1020210122012 A KR 1020210122012A KR 20210122012 A KR20210122012 A KR 20210122012A KR 20230039105 A KR20230039105 A KR 20230039105A
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- skin
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/49—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing heterocyclic compounds
- A61K8/4973—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing heterocyclic compounds with oxygen as the only hetero atom
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
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- A—HUMAN NECESSITIES
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- A61P17/00—Drugs for dermatological disorders
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
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- A61Q19/02—Preparations for care of the skin for chemically bleaching or whitening the skin
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- A—HUMAN NECESSITIES
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- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2200/00—Function of food ingredients
- A23V2200/30—Foods, ingredients or supplements having a functional effect on health
- A23V2200/318—Foods, ingredients or supplements having a functional effect on health having an effect on skin health and hair or coat
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
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Abstract
Description
본 발명은 잔사틴(Xanthatin; CAS No. 26791-73-1) 을 이용한 피부 미백용 조성물에 관한 것이다. The present invention is Xanthatin (CAS No. 26791-73-1) It relates to a composition for skin whitening using
현대사회에서 생활수준이 향상됨에 따라 건강뿐만 아니라 미용에 대한 관심도 점차 높아지고 있다. 이 중 자외선, 대기오염 등에 의한 피부손상 및 노화로 인하여 유발되는 기미(melasma), 주근깨(freckle)와 같은 색소 침착형 피부 질환 개선과 전반적인 피부톤 개선을 위한 제품 개발이 활발하다. As living standards improve in modern society, interest in beauty as well as health is gradually increasing. Among them, development of products for improvement of pigmentation-type skin diseases such as melasma and freckles caused by skin damage and aging caused by ultraviolet rays, air pollution, etc., and improvement of overall skin tone is active.
표피의 기저층에 존재하는 멜라닌(melanin)은 피부색을 결정하는 주요한 색소로서 자외선에 의한 손상을 막고 활성산소를 제거하여 피부를 보호역할을 한다. 그러나 멜라닌이 국소적으로 과생성 될 경우 기미, 주근깨, 검은반점, 피부암 등의 피부 질환을 일으키게 된다. 또한 지속적으로 과생성 된 멜라닌이 진피에 머무르게 되면 피부색을 검게 변하게 하거나 안색을 칙칙하게 바꾸기 때문에 미백에 대한 욕구를 유도하게 된다(Pigment Cell Res. 2007 Feb;20(1):2-13.; Curr Biol. 2011 Nov 22;21(22):1906-11.;Ann Dermatol Venereol. 2012 Nov;139 Suppl 3:S73-7).Melanin present in the basal layer of the epidermis is a major pigment that determines skin color and plays a role in protecting the skin by preventing damage caused by ultraviolet rays and removing active oxygen. However, when melanin is locally overproduced, it causes skin diseases such as melasma, freckles, dark spots, and skin cancer. In addition, when continuously overproduced melanin stays in the dermis, it turns the skin color black or dull, leading to a desire for whitening (Pigment Cell Res. 2007 Feb;20(1):2-13.; Curr Biol. 2011 Nov. 22;21(22):1906-11.; Ann Dermatol Venereol.
멜라닌은 생물체에 널리 존재하는 페놀류 고분자 천연 색소로 인체에서는 표피 기저층에 있는 멜라노사이트(melanocyte) 내의 멜라노솜(melanosome)에서 합성되며 주변 각질 세포로 전이되어 사람의 피부색을 나타낸다. 멜라닌이 비정상적으로 적게 되면 백반증과 같은 피부 병변이 유발되며, 반대로 과잉으로 생산될 경우에는 흑피증(melasma), 염증후흑피증(postinflammatory melanoderma), 일광흑색점(solar lentigo) 등과 같은 후천성 과다색소침착증을 포함하는 다양한 피부질환이 유발될 수 있다.Melanin is a phenolic polymeric natural pigment widely present in living organisms. In the human body, it is synthesized in melanosomes in melanocytes in the basal layer of the epidermis, and is transferred to surrounding keratinocytes to show human skin color. When melanin is abnormally low, skin lesions such as vitiligo are induced, and on the contrary, when it is produced in excess, acquired hyperpigmentation such as melasma, postinflammatory melanoderma, and solar lentigo A variety of skin diseases, including
멜라닌의 합성은 피부의 기저층에 존재하는 멜라노사이트(melanocyte)에서 멜라닌소체(melanosome)를 만들어 이루어지며 멜라닌이 합성되는 일련의 과정을 총칭하여 멜라닌 생성 경로(melanogenesis)라 한다. 멜라닌 생성은 아미노산의 하나인 티로신(tyrosine)을 기질로 하며, 티로시나아제(tyrosinase), TRP-1(tyrosinase related protein-1), TRP-2(tyrosinase related protein-2)에 의해 DOPA(3,4-dihydroxyphenylalanine)를 거쳐 도파 퀴논(DOPA quinone)으로 전환된 후 비효소적 반응, 자발적 산화 과정을 거친 후 아미노산 혹은 단백질과의 중합 반응에 의해 멜라닌이 합성된다(Korean J. FOOD SCI. TECHNOL., 2000. 32(3):736; Journal of Life Science, 2013. 23(12):1445; Pigment Cell Res.1999. 12(1):4-12.). 최근에는 티로시나아제, TRP-1, TRP-2뿐만 아니라 MITF(microphtalmia associated transcription factor) 등과 같은 단백질을 이용하여 멜라닌 합성 억제에 대한 연구도 이루어지고 있다. 주요한 세포 내 신호 전달 경로는 cAMP/PKA(cyclic monophosphate/protein kinase A) 경로로서, cAMP는 PKA, CREB1(cAMP responsive element binding protein 1)을 경유하여 MITF의 발현을 촉진하며 MITF는 멜라닌 합성 과정에서 중요한 전사 조절 인자로 티로시나아제, TRP-1, TRP-2의 전사를 촉진한다(Pharmazie. 2015 Oct;70(10):646-9.; Int J Mol Sci. 2015 Oct 13;16(10):24219-42; Curr Pharm Biotechnol. 2015;16(12):1111-9.). The synthesis of melanin is achieved by making melanosomes in melanocytes present in the basal layer of the skin, and a series of processes in which melanin is synthesized are collectively referred to as melanogenesis. Melanin production uses tyrosine, one of the amino acids, as a substrate, and DOPA (3, After conversion to DOPA quinone through 4-dihydroxyphenylalanine), melanin is synthesized by polymerization with amino acids or proteins after undergoing a non-enzymatic reaction and spontaneous oxidation process (Korean J. FOOD SCI. TECHNOL., 2000. 32(3):736;Journal of Life Science, 2013.23(12):1445;Pigment Cell Res.1999.12(1):4-12.). Recently, studies on the inhibition of melanin synthesis have been conducted using proteins such as tyrosinase, TRP-1 and TRP-2 as well as microphtalmia associated transcription factor (MITF). The major intracellular signal transduction pathway is the cAMP/PKA (cyclic monophosphate/protein kinase A) pathway. cAMP promotes the expression of MITF via PKA and CREB1 (cAMP responsive element binding protein 1), and MITF plays an important role in melanin synthesis. As a transcriptional regulator, it promotes the transcription of tyrosinase, TRP-1 and TRP-2 (Pharmazie. 2015 Oct;70(10):646-9.; Int J Mol Sci. 2015 Oct 13;16(10): 24219-42; Curr Pharm Biotechnol. 2015;16(12):1111-9.).
멜라닌을 생성하는 첫 단계로 티로시나아제가 가장 중요한 역할을 하기 때문에 미백제에 대한 연구는 주로 티로시나아제(tyrosinase) 활성을 조절하기 위하여 티로시나아제 활성 저해 물질이나 티로시나아제의 기질에 대한 길항물질을 개발하는 방향으로 연구가 진행되어 왔다(Kor. J. Pharmacogn., 44(3):220, 2013).Since tyrosinase plays the most important role in the first step of melanin production, studies on whitening agents are mainly focused on tyrosinase activity inhibitors or antagonists for tyrosinase substrates to regulate tyrosinase activity. Research has been conducted in the direction of developing (Kor. J. Pharmacogn., 44(3):220, 2013).
현재 미백제로 가장 많이 사용되는 알부틴(arbutin)은 티로신의 길항제이며, 코지산(kojic acid)은 티로시나아제 활성 부위를 킬레이트화하여 티로신에서 도파, 도파퀴논으로 진행되는 과정을 저해한다(Korean J. Plant Res., 26(5):526, 2013; J. Korean Soc. Food Sci. Nutr., 41(2):156, 2012). 알부틴과 코직산은 강력한 미백효과가 있는 것으로 알려져 있으나, 피부 자극 등의 일부 부작용으로 인하여 상대적으로 부작용이 적은 천연의 물질로부터 미백 활성을 가지는 물질을 찾기 위한 연구가 활발하게 이루어지고 있다.Arbutin, the most commonly used whitening agent, is a tyrosine antagonist, and kojic acid chelates the active site of tyrosinase to inhibit the process of tyrosine to dopa and dopaquinone (Korean J. Plant Res., 26(5):526, 2013; J. Korean Soc. Food Sci. Nutr., 41(2):156, 2012). Although arbutin and kojic acid are known to have strong whitening effects, due to some side effects such as skin irritation, studies are being actively conducted to find substances having whitening activity from natural substances with relatively few side effects.
본 발명은 천연물로부터 분리·동정한 잔사틴의 미백 활성을 개시한다.The present invention discloses the whitening activity of zansatin isolated and identified from natural products.
본 발명의 목적은 잔사틴을 이용한 피부 미백용 조성물을 제공하는 데 있다.An object of the present invention is to provide a composition for skin whitening using zansatin.
본 발명의 다른 목적이나 구체적인 목적은 이하에서 제시될 것이다.Other objects or specific objects of the present invention will be presented below.
본 발명은 아래의 실시예 및 실험예에서 확인되는 바와 같이, 아래 화학식 1의 잔사틴이 마우스 유래 멜라노마 세포주인 B16F10에 처리될 때, 농도 의존적으로 멜라닌 생성을 저해하고 또한 티로시나아제, TRP-1 및 TRP-2 단백질의 발현을 억제함으로써 완성된 것이다.As confirmed in the following examples and experimental examples, the present invention inhibits melanin production in a concentration-dependent manner and also inhibits tyrosinase, TRP- 1 and TRP-2 proteins were inhibited.
<화학식 1><
전술한 바를 고려할 때, 본 발명의 피부 미백용 조성물은 잔사틴을 유효성분으로 포함함을 특징으로 한다.Considering the foregoing, the skin whitening composition of the present invention is characterized in that it contains zansatin as an active ingredient.
본 명세서에서 "유효성분"이란 단독으로 목적하는 활성을 나타내거나 또는 그 자체는 활성이 없는 담체와 함께 활성을 나타낼 수 있는 성분을 의미한다.In the present specification, "active ingredient" means a component that exhibits the desired activity alone or can exhibit activity in combination with a carrier having no activity itself.
본 발명의 조성물은 그 유효성분을 제품, 제형 등에 따라 피부 미백 활성 등을 나타낼 수 있는 한 임의의 양(유효량)으로 포함할 수 있는데, 통상적인 유효량은 조성물 전체 중량을 기준으로 할 때 0.001 중량 % 내지 15 중량 % 범위 내에서 결정될 것이다. 여기서 "유효량"이란 그 적용 대상인 포유동물 바람직하게는 사람에게 의료 전문가, 피부 전문가 등의 제언에 의한 투여 기간 동안 본 발명의 조성물이 투여될 때, 피부 미백 효과 등 의도한 의료적·피부학적 효과를 나타낼 수 있는, 본 발명의 조성물에 포함되는 유효성분의 양을 말한다. 이러한 유효량은 당업자의 통상의 능력 범위 내에서 실험적으로 결정될 수 있다.The composition of the present invention may contain the active ingredient in any amount (effective amount) as long as it can exhibit skin whitening activity, etc., depending on the product, formulation, etc., and a typical effective amount is 0.001% by weight based on the total weight of the composition. to 15% by weight. Here, "effective amount" means that when the composition of the present invention is administered to a mammal, preferably a human, during the administration period according to the advice of medical experts, skin experts, etc., the intended medical and dermatological effects such as skin whitening effect It refers to the amount of the active ingredient contained in the composition of the present invention that can be expressed. Such an effective amount can be determined empirically within the ordinary skill of the skilled artisan.
본 발명의 조성물은 유효성분 이외에, 피부 미백 활성의 상승·보강 등을 위하여 또는 피부 주름 개선 활성, 피부 과민 반응 억제 활성, 피부 보호 활성(자외선에 의한 피부 손상 억제, 피부 보습 등) 등 유사활성의 부가를 통한 복용이나 섭취, 사용의 편리성을 증진시키기 위하여, 당업계에서 이미 안전성이 검증되고 해당 활성을 갖는 것으로 공지된 임의의 화합물이나 천연 추출물을 추가로 포함할 수 있다.In addition to the active ingredients, the composition of the present invention has similar activities such as skin whitening activity, skin wrinkle improvement activity, skin hypersensitivity reaction inhibition activity, skin protection activity (inhibition of skin damage by ultraviolet rays, skin moisturizing, etc.) In order to enhance the convenience of intake, intake, and use through addition, any compound or natural extract known to have a corresponding activity and whose safety has already been verified in the art may be further included.
이러한 화합물 또는 추출물에는 각국 약전(한국에서는 "대한민국약전"), 각국 건강기능식품공전(한국에서는 식약처 고시인 "건강기능식품 기준 및 규격"임), 각국 기능성화장품공전(한국에서는 식약처 고시인 "기능성화장품 기준 및 시험방법") 등의 공정서에 실려 있는 화합물 또는 추출물, 의약품의 제조·판매를 규율하는 각국의 법률(한국에서는 "약사법"임)에 따라 품목 허가를 받은 화합물 또는 추출물, 건강기능식품의 제조·판매를 규율하는 각국 법률(한국에서는 「건강기능식품에관한법률」임)에 따라 기능성이 인정된 화합물 또는 추출물, 기능성 화장품의 제조·판매를 규율하는 각국 법률(한국에서는 「화장품법」임)에 따라 기능성이 인정된 화합물 또는 추출물이 포함된다. 예컨대 피부 미백 성분으로서는 알부틴, 나이아신아마이드, 아스코빌글루코사이드, 알파-비사보롤, 유용성 감초 추출물(Oil Soluble Licorice(Glycyrrhiza) Extract) 등을 들 수 있으며, 또 피부 주름 개선 성분으로서는 레티놀, 레티닐팔미테이트, 아데노신, 폴리에톡실레이티드아마이드 등을 들 수 있고, 자외선 보호 성분으로서는 드로메트리졸, 드로메트리졸트리실록산, 디갈로일트리올리에이트, 디메치코디에칠벤잘말로네이트, 디에칠헥실부타미도트리아존, 소나무 껍질 추출물 등의 복합물, 포스파티딜세린, 핑거루트 추출 분말, 홍삼과 산수유 등의 복합 추출물 등을 들 수 있다. 또 보습 성분으로서는 AP 콜라겐 효소 분해 펩타이드, Collactive 콜라겐 펩타이드, N-아세틸글루코사민, 곤약 감자 추출물, 민들레 등의 복합 추출물, 쌀겨 추출물, 옥수수 배아 추출물, 저분자 콜라겐 펩타이드, 지초 추출 분말, 포스파티딜세린, 히알루론산 등을 들 수 있으며, '과민피부상태 개선' 기능성 성분으로서는 L. sakei Probio 65, 감마리놀렌산 함유 유지, 과채 유래 유산균인 L.plantarum CJLP133, 프로바이오틱스 ATP 등을 들 수 있다. 이러한 화합물 또는 천연 추출물은 본 발명의 조성물에 그 유효성분과 함께 하나 이상 포함될 수 있다.These compounds or extracts include each country's Pharmacopoeia ("Korean Pharmacopoeia" in Korea), each country's Health Functional Food Codex (in Korea, it is "Health Functional Food Standards and Specifications", a notification by the Ministry of Food and Drug Safety), and each country's Functional Cosmetics Codex (in Korea, a notification by the Ministry of Food and Drug Safety Compounds or extracts listed in compendial documents such as "Functional Cosmetics Standards and Test Methods"), compounds or extracts approved as items under the laws of each country governing the manufacture and sale of pharmaceuticals ("Pharmaceutical Affairs Act" in Korea), health In accordance with the laws of each country regulating the manufacture and sale of functional foods (in Korea, it is the 「Health Functional Food Act」), the laws of each country regulating the manufacture and sale of compounds or extracts with recognized functionality, and the manufacture and sale of functional cosmetics (in Korea, the 「Cosmetics Act」 ”), compounds or extracts whose functionality has been recognized are included. For example, skin whitening ingredients include arbutin, niacinamide, ascorbyl glucoside, alpha-bisabolol, oil soluble licorice (Glycyrrhiza) Extract, etc., and skin wrinkle improvement ingredients include retinol and retinyl palmitate. , adenosine, polyethoxylated amide, and the like, and examples of ultraviolet protection components include drometrizole, drometrizoletrisiloxane, digaloyltrioleate, dimethicodyethylbenzalmalonate, and diethylhexylbutamido. complexes such as triazone and pine bark extract, phosphatidylserine, finger root extract powder, and complex extracts such as red ginseng and cornus officinalis. In addition, as moisturizing ingredients, AP collagen enzyme decomposition peptide, Collactive collagen peptide, N-acetylglucosamine, konjac potato extract, complex extracts such as dandelion, rice bran extract, corn germ extract, low molecular weight collagen peptide, weed extract powder, phosphatidylserine, hyaluronic acid, etc. Functional ingredients for 'improving sensitive skin conditions' include L. sakei Probio 65, gamma-linolenic acid-containing oil, L.plantarum CJLP133, a lactic acid bacterium derived from fruits and vegetables, and probiotics ATP. One or more of these compounds or natural extracts may be included in the composition of the present invention together with the active ingredient.
본 발명의 피부 미백용 조성물은 구체적인 양태에 있어서, 화장료 조성물로 파악할 수 있다.In a specific embodiment, the composition for skin whitening of the present invention can be regarded as a cosmetic composition.
본 발명의 조성물이 화장료 조성물로 파악될 경우 그 화장료 조성물은 그 용도상, 법률상 임의의 제품 구분을 띨 수 있으며, 구체적으로 피부 미백 등의 용도를 가진 기능성 화장품, 비기능성 일반 화장품 등일 수 있다. 제품 형태에 있어서도 임의의 제품 형태를 띨 수 있는데, 구체적으로 용액, 현탁액, 유탁액, 페이스트, 젤, 크림, 로션, 파우더, 비누, 계면활성제-함유 클렌징, 오일, 분말 파운데이션, 유탁액 파운데이션, 왁스 파운데이션, 스프레이 등으로 제형화될 수 있다. 구체적인 제품 형태에 있어서는 유연 화장수, 영양 화장수, 영양 크림, 마사지 크림, 에센스, 아이 크림, 클렌징 크림, 클렌징 포옴, 클렌징 워터, 팩, 스프레이 또는 파우더의 제형 등일 수 있다.When the composition of the present invention is recognized as a cosmetic composition, the cosmetic composition may be classified as an arbitrary product in terms of its use and law, and specifically, may be a functional cosmetic having a purpose such as skin whitening, a non-functional general cosmetic, and the like. In product form, it may take any product form, specifically, solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, surfactant-containing cleansing, oil, powder foundation, emulsion foundation, wax It can be formulated as a foundation, spray, and the like. In a specific product form, it may be a softening lotion, nutrient lotion, nutrient cream, massage cream, essence, eye cream, cleansing cream, cleansing foam, cleansing water, pack, spray or powder formulation.
본 발명의 화장료 조성물은 그 유효성분 이외에 화장료 조성물에 통상적으로 이용되는 성분들, 예컨대, 안정화제, 용해화제, 계면활성제, 비타민, 색소 및 항료와 같은 통상적인 보조제, 및 담체를 포함할 수 있다. The cosmetic composition of the present invention may include, in addition to the active ingredient, ingredients commonly used in cosmetic compositions, for example, stabilizers, solubilizers, surfactants, vitamins, conventional adjuvants such as pigments and fragrances, and carriers.
본 발명의 제형이 페이스트, 크림 또는 젤인 경우에는 담체 성분으로서 동물성유, 식물성유, 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 oil, vegetable oil, wax, paraffin, starch, tracanth, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide may be used as a carrier component. can
본 발명의 제형이 파우더 또는 스프레이인 경우에는 담체 성분으로서 락토스, 탈크, 실리카, 알루미늄 히드록시드, 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, additional chlorofluorohydrocarbon, propane / May contain a propellant such as butane or dimethyl ether.
본 발명의 제형이 용액 또는 유탁액인 경우에는 담체 성분으로서 용매, 용해화제 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, 소르비탄의 지방산 에스테르 등이 이용될 수 있다.When the formulation of the present invention is a solution or emulsion, a solvent, solubilizing 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 ,3-butyl glycol oil, glycerol aliphatic esters, polyethylene glycol, fatty acid esters of sorbitan, and the like can be used.
본 발명의 제형이 현탁액인 경우에는 담체 성분으로서 물, 에탄올 또는 프로필렌 글리콜과 같은 액상의 희석제, 에톡실화 이소스테아릴 알코올, 폴리옥시에틸렌 소르비톨 에스테르, 폴리옥시에틸렌 소르비탄 에스테르와 같은 현탁제, 미소결정성 셀룰로오스, 알루미늄 메타히드록시드, 벤토나이트, 아가 등이 이용될 수 있다.When the formulation of the present invention is a suspension, as a carrier component, a liquid diluent such as water, ethanol or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester, or polyoxyethylene sorbitan ester, microcrystals Star cellulose, aluminum metahydroxide, bentonite, agar, and the like 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, 라놀린 유도체 또는 에톡실화 글리세롤 지방산 에스테르 등이 이용될 수 있다.When the formulation of the present invention is surfactant-containing cleansing, as carrier components, aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyl taurate, sarcosinate, fatty acid amide Ether sulfates, alkylamidobetaines, aliphatic alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, lanolin derivatives or ethoxylated glycerol fatty acid esters and the like can be used.
본 발명의 화장료 조성물은 피부 미백 활성을 나타내는 그 유효성분을 포함하는 것을 제외하고는 당업계에 통상적으로 행하여지는 화장료 조성물의 제조방법에 따라 제조할 수 있다.The cosmetic composition of the present invention may be prepared according to a method for preparing a cosmetic composition conventionally performed in the art, except for including an active ingredient exhibiting skin whitening activity.
본 발명의 조성물은 구체적인 양태에 있어서, 식품 조성물로서 파악할 수 있다. In a specific aspect, the composition of the present invention can be regarded as a food composition.
본 발명의 식품 조성물은 어떠한 형태로도 제조될 수 있으며, 예컨대 차, 쥬스, 탄산음료, 이온음료 등의 음료류, 우유, 요구르트 등의 가공 유류, 껌류, 떡, 한과, 빵, 과자, 면 등의 식품류, 정제, 캡슐, 환, 과립, 액상, 분말, 편상, 페이스트상, 시럽, 겔, 젤리, 바 등의 건강기능식품 제제류 등으로 제조될 수 있다. The food composition of the present invention can be prepared in any form, for example, beverages such as tea, juice, carbonated beverages, and ionic beverages, processed oils such as milk and yogurt, chewing gum, rice cakes, traditional Korean snacks, bread, snacks, noodles, etc. It can be manufactured into health functional food preparations such as foods, tablets, capsules, pills, granules, liquid, powder, flakes, pastes, syrups, gels, jellies, and bars.
또 본 발명의 식품 조성물은 법률상·기능상의 구분에 있어서 제조·유통 시점의 시행 법규에 부합하는 한 임의의 제품 구분을 띨 수 있다. 예컨대 한국 「건강기능식품에관한법률」에 따른 건강기능식품이거나, 한국 「식품위생법」의 식품공전(식약처 고시 「식품의 기준 및 규격」)상 각 식품유형에 따른 과자류, 두류, 다류, 음료류, 특수용도식품 등일 수 있다.In addition, the food composition of the present invention may take any product classification as long as it conforms to the enforcement regulations at the time of manufacture and distribution in legal and functional classification. For example, health functional food according to the Korean 「Health Functional Food Act」, or snacks, legumes, teas, beverages according to each food type according to the Food Code (MFDS notification 「Food Standards and Specifications」) of the Korea 「Food Sanitation Act」 , special purpose foods, etc.
본 발명의 식품 조성물에는 그 유효성분 이외에 식품첨가물이 포함될 수 있다. 식품첨가물은 일반적으로 식품을 제조, 가공 또는 보존함에 있어 식품에 첨가되어 혼합되거나 침윤되는 물질로서 이해될 수 있는데, 식품과 함께 매일 그리고 장기간 섭취되므로 그 안전성이 보장되어야 한다. 식품의 제조·유통을 규율하는 각국 법률(한국에서는 「식품위생법」임)에 따른 식품첨가물공전에는 안전성이 보장된 식품첨가물이 성분 면에서 또는 기능 면에서 한정적으로 규정되어 있다. 한국 식품첨가물공전(식약처 고시 「식품첨가물 기준 및 규격」)에서는 식품첨가물이 성분 면에서 화학적 합성품, 천연 첨가물 및 혼합 제제류로 구분되어 규정되어 있는데, 이러한 식품첨가물은 기능 면에 있어서는 감미제, 풍미제, 보존제, 유화제, 산미료, 점증제 등으로 구분된다. The food composition of the present invention may include food additives in addition to the active ingredient. Food additives may generally be understood as substances that are added to, mixed with, or infiltrated into food in manufacturing, processing, or preserving food. Since food is consumed daily and for a long period of time, its safety must be guaranteed. According to the Food Additives Code under the laws of each country governing the manufacture and distribution of food (in Korea, it is the 「Food Sanitation Act」), safety-guaranteed food additives are limitedly regulated in terms of ingredients or functions. In the Korean Food Additives Codex (MFDS notification 「Standards and Specifications for Food Additives」), food additives are classified into chemical synthetic products, natural additives and mixed preparations in terms of ingredients and are regulated. It is divided into additives, preservatives, emulsifiers, acidulants, and thickeners.
감미제는 식품에 적당한 단맛을 부여하기 위하여 사용되는 것으로, 천연의 것이거나 합성된 것 모두 본 발명의 조성물에 사용할 수 있다. 바람직하게는 천연 감미제를 사용하는 경우인데, 천연 감미제로서는 옥수수 시럽 고형물, 꿀, 수크로오스, 프룩토오스, 락토오스, 말토오스 등의 당 감미제를 들 수 있다. Sweeteners are used to impart appropriate sweetness to foods, and both natural and synthetic sweeteners can be used in the composition of the present invention. Preferably, a natural sweetener is used, and examples of the natural sweetener include sugar sweeteners such as corn syrup solids, honey, sucrose, fructose, lactose, and maltose.
풍미제는 맛이나 향을 좋게 하기 위하여 사용될 수 있는데, 천연의 것과 합성된 것 모두 사용될 수 있다. 바람직하게는 천연의 것을 사용하는 경우이다. 천연의 것을 사용할 경우에 풍미 이외에 영양 강화의 목적도 병행할 수 있다. 천연 풍미제로서는 사과, 레몬, 감귤, 포도, 딸기, 복숭아 등에서 얻어진 것이거나 녹차잎, 둥굴레, 대잎, 계피, 국화 잎, 자스민 등에서 얻어진 것일 수 있다. 또 인삼(홍삼), 죽순, 알로에 베라, 은행 등에서 얻어진 것을 사용할 수 있다. 천연 풍미제는 액상의 농축액이나 고형상의 추출물일 수 있다. 경우에 따라서 합성 풍미제가 사용될 수 있는데, 합성 풍미제는 에스테르, 알콜, 알데하이드, 테르펜 등이 이용될 수 있다. Flavors may be used to improve taste or aroma, and both natural and synthetic flavors may be used. Preferably, it is the case of using a natural one. In case of using natural ones, in addition to flavor, the purpose of enhancing nutrition can also be combined. As a natural flavoring agent, it may be obtained from apples, lemons, tangerines, grapes, strawberries, peaches, etc., or obtained from green tea leaves, roundworms, bamboo leaves, cinnamon, chrysanthemum leaves, jasmine, and the like. In addition, those obtained from ginseng (red ginseng), bamboo shoots, aloe vera, ginkgo, etc. can be used. Natural flavors can be liquid concentrates or solid extracts. In some cases, synthetic flavors may be used, and synthetic flavors may include esters, alcohols, aldehydes, terpenes, and the like.
보존제로서는 소듐 소르브산칼슘, 소르브산나트륨, 소르브산칼륨, 벤조산칼슘, 벤조산나트륨, 벤조산칼륨, EDTA(에틸렌디아민테트라아세트산) 등이 사용될 수 있고, 또 유화제로서는 아카시아검, 카르복시메틸셀룰로스, 잔탄검, 펙틴 등을 들 수 있으며, 산미료로서는 연산, 말산, 푸마르산, 아디프산, 인산, 글루콘산, 타르타르산, 아스코르브산, 아세트산, 인산 등이 사용될 수 있다. 산미료는 맛을 증진시키는 목적 이외에 미생물의 증식을 억제할 목적으로 식품 조성물이 적정 산도로 되도록 첨가될 수 있다.As preservatives, sodium sorbate, sodium sorbate, potassium sorbate, calcium benzoate, sodium benzoate, potassium benzoate, EDTA (ethylenediaminetetraacetic acid), etc. may be used, and as emulsifiers, acacia gum, carboxymethylcellulose, xanthan gum, pectin and the like, and examples of the acidulant include acid salt, malic acid, fumaric acid, adipic acid, phosphoric acid, gluconic acid, tartaric acid, ascorbic acid, acetic acid, phosphoric acid, and the like. Acidulants may be added to the food composition to have an appropriate acidity for the purpose of inhibiting the growth of microorganisms in addition to improving taste.
점증제로서는 현탁화 구현제, 침강제, 겔형성제, 팽화제 등이 사용될 수 있다.As the thickening agent, a suspending agent, a sedimentation agent, a gel forming agent, a swelling agent, and the like may be used.
본 발명의 식품 조성물은 전술한 바의 식품첨가물 이외에, 기능성과 영양성을 보충, 보강할 목적으로 당업계에 공지되고 식품첨가물로서 안정성이 보장된 생리활성 물질이나 미네랄류를 포함할 수 있다.In addition to the food additives described above, the food composition of the present invention may include physiologically active substances or minerals known in the art and whose stability is guaranteed as food additives for the purpose of supplementing or reinforcing functionality and nutrition.
그러한 생리활성 물질로서는 녹차 등에 포함된 카테킨류, 비타민 B1, 비타민 C, 비타민 E, 비타민 B12 등의 비타민류, 토코페롤, 디벤조일티아민 등을 들 수 있으며, 미네랄류로서는 구연산 칼슘 등의 칼슘 제제, 스테아린산마그네슘 등의 마그네슘 제제, 구연산철 등의 철 제제, 염화 크롬, 요오드칼륨, 셀레늄, 게르마늄, 바나듐, 아연 등을 들 수 있다. Examples of such physiologically active substances include catechins contained in green tea, vitamins such as vitamin B1, vitamin C, vitamin E, and vitamin B12, tocopherol, dibenzoylthiamine, and the like. Examples of minerals include calcium preparations such as calcium citrate and magnesium stearate. magnesium preparations such as the like, iron preparations such as iron citrate, chromium chloride, potassium iodine, selenium, germanium, vanadium, zinc, and the like.
본 발명의 식품 조성물에는 전술한 바의 식품첨가물이 제품 유형에 따라 그 첨가 목적을 달성할 수 있는 적량으로 포함될 수 있다.The food composition of the present invention may include the above-mentioned food additives in an appropriate amount to achieve the purpose of addition according to the type of product.
본 발명의 식품 조성물에 포함될 수 있는 기타의 식품첨가물과 관련하여서는 각국 식품공전이나 식품첨가물 공전을 참조할 수 있다.Regarding other food additives that may be included in the food composition of the present invention, reference may be made to national food codes or food additive codes.
본 발명의 피부 미백용 조성물은 구체적인 양태에 있어서, 약제학적 조성물로 파악할 수 있다. In a specific aspect, the composition for skin whitening of the present invention can be understood as a pharmaceutical composition.
본 발명의 피부 미백용 조성물이 약제학적 조성물로 파악될 경우 대상 질환은 피부 과색소침착증으로 이해될 수 있으며, 그러한 과색소침작증의 구체적인 예로서는 주근깨, 노인성 반점, 간반, 기미, 갈색 또는 흑점, 일광 색소반, 푸른 흑피증, 칼슘 길항제와 같은 약물 사용 후의 과다색소침착, 조직경화치료법에 따른 과다색소침착 후유증, 임신성 갈색반, 경구 피임약을 복용한 여성에서의 흑피증, 찰상 및 화상을 비롯한 상처 또는 피부염으로 인한 염증 후 과다 색소 침착, 광독성 반응 또는 다른 유사한 소형의 고정 색소성 병변 등이 포함될 수 있으나, 이에 제한되는 것은 아니다.When the composition for skin whitening of the present invention is understood as a pharmaceutical composition, the target disease can be understood as skin hyperpigmentation, and specific examples of such hyperpigmentation include freckles, senile spots, liver spots, melasma, brown or dark spots, solar pigment Melasma blue, hyperpigmentation after the use of drugs such as calcium antagonists, hyperpigmentation sequelae following tissue sclerotherapy, brown spots of pregnancy, melasma in women taking oral contraceptives, wounds or dermatitis including scratches and burns may include, but are not limited to, post-inflammatory hyperpigmentation, phototoxic reaction, or other similar small fixed pigmented lesions.
본 발명의 약제학적 조성물은 유효성분 이외에 약제학적으로 허용되는 담체를 포함하여 당업계에 공지된 통상의 방법으로 투여 경로에 따라 경구용 제형 또는 비경구용 제형으로 제조될 수 있다. 여기서 투여 경로는 국소 경로, 경구 경로, 정맥 내 경로, 근육 내 경로, 및 점막 조직을 통한 직접 흡수를 포함하는 임의의 적절한 경로일 수 있으며, 두 가지 이상의 경로를 조합하여 사용할 수도 있다. 두 가지 이상 경로의 조합의 예는 투여 경로에 따른 두 가지 이상의 제형의 약물이 조합된 경우로서 예컨대 1차로 어느 한 약물은 정맥 내 경로로 투여하고 2차로 다른 약물은 국소 경로로 투여하는 경우이다. The pharmaceutical composition of the present invention may be formulated into an oral formulation or a parenteral formulation according to the route of administration by a conventional method known in the art, including a pharmaceutically acceptable carrier in addition to the active ingredient. The route of administration herein may be any appropriate route including topical route, oral route, intravenous route, intramuscular route, and direct absorption through mucosal tissue, and two or more routes may be used in combination. An example of a combination of two or more routes is a case in which two or more formulations of drugs are combined according to the route of administration, for example, one drug is firstly administered intravenously and the other drug is secondly administered through a topical route.
약학적으로 허용되는 담체는 투여 경로나 제형에 따라 당업계에 주지되어 있으며, 구체적으로는 "대한민국약전"을 포함한 각국의 약전을 참조할 수 있다. Pharmaceutically acceptable carriers are well known in the art depending on the route of administration or dosage form, and specifically, reference may be made to the pharmacopoeia of each country including the "Korean Pharmacopoeia".
본 발명의 약제학적 조성물이 경구용 제형으로 제조될 경우, 적합한 담체와 함께 당업계에 공지된 방법에 따라 분말, 과립, 정제, 환제, 당의정제, 캡슐제, 액제, 겔제, 시럽제, 현탁액, 웨이퍼 등의 제형으로 제조될 수 있다. 이때 적합한 담체의 예로서는 락토스, 글루코스, 슈크로스, 덱스트로스, 솔비톨, 만니톨, 자일리톨 등의 당류, 옥수수 전분, 감자 전분, 밀 전분 등의 전분류, 셀룰로오스, 메틸셀룰로오스, 에틸셀룰로오스, 나트륨 카르복시메틸셀룰로오스, 하이드록시프로필메틸셀룰로오스 등의 셀룰로오스류, 폴리비닐 피롤리돈, 물, 메틸히드록시벤조에이트, 프로필히드록시벤조에이트, 마그네슘 스테아레이트, 광물유, 맥아, 젤라틴, 탈크, 폴리올, 식물성유, 에탄올, 그리세롤 등을 들 수 있다. 제제화활 경우 필요에 따라적절한 결합제, 윤활제, 붕해제, 착색제, 희석제 등을 포함시킬 수 있다. 적절한 결합제로서는 전분, 마그네슘 알루미늄 실리케이트, 전분페리스트, 젤라틴, 메틸셀룰로스, 소듐 카복시메틸셀룰로스, 폴리비닐피롤리돈, 글루코스, 옥수수 감미제, 소듐 알지네이트, 폴리에틸렌 글리콜, 왁스 등을 들 수 있고, 윤활제로서는 올레산나트륨, 스테아르산나트륨, 스테아르산마그네슘, 벤조산나트륨, 초산나트륨, 염화나트륨, 실리카, 탈쿰, 스테아르산, 그것의 마그네슘염과 칼슘염, 폴리데틸렌글리콜 등을 들 수 있으며, 붕해제로서는 전분, 메틸 셀룰로스, 아가(agar), 벤토나이트, 잔탄 검, 전분, 알긴산 또는 그것의 소듐 염 등을 들 수 있다. 또 희석제로서는 락토즈, 덱스트로즈, 수크로즈, 만니톨, 소비톨, 셀룰로스, 글라이신 등을 들 수 있다. When the pharmaceutical composition of the present invention is prepared as an oral dosage form, powder, granule, tablet, pill, dragee, capsule, liquid, gel, syrup, suspension, wafer according to a method known in the art together with a suitable carrier. It can be prepared in formulations such as Examples of suitable carriers include sugars such as lactose, glucose, sucrose, dextrose, sorbitol, mannitol, and xylitol, starches such as corn starch, potato starch, and wheat starch, cellulose, methylcellulose, ethylcellulose, sodium carboxymethylcellulose, Celluloses such as hydroxypropylmethylcellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, magnesium stearate, mineral oil, malt, gelatin, talc, polyol, vegetable oil, ethanol, Serol etc. are mentioned. In the case of formulation, appropriate binders, lubricants, disintegrants, coloring agents, diluents, etc. may be included as needed. Suitable binders include starch, magnesium aluminum silicate, starch ferrite, gelatin, methylcellulose, sodium carboxymethylcellulose, polyvinylpyrrolidone, glucose, corn sweetener, sodium alginate, polyethylene glycol, wax, and the like, and oleic acid as a lubricant Sodium, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride, silica, talcum, stearic acid, magnesium salts and calcium salts thereof, polyethylene glycol, etc. may be mentioned. As disintegrants, starch, methyl cellulose , agar, bentonite, xanthan gum, starch, alginic acid or its sodium salt, and the like. Examples of the diluent include lactose, dextrose, sucrose, mannitol, sorbitol, cellulose, and glycine.
본 발명의 약제학적 조성물이 비경구용 제형으로 제조될 경우, 적합한 담체와 함께 당업계에 공지된 방법에 따라 주사제, 경피 투여제, 비강 흡입제 및 좌제의 형태로 제제화될 수 있다. 주사제로 제제화할 경우 적합한 담체로서는 수성 등장 용액 또는 현탁액을 사용할 수 있으며, 구체적으로는 트리에탄올 아민이 함유된 PBS(phosphate buffered saline)나 주사용 멸균수, 5% 덱스트로스 같은 등장 용액 등을 사용할 수 있다. 경피 투여제로 제제화할 경우 연고제, 크림제, 로션제, 겔제, 외용액제, 파스타제, 리니멘트제, 에어롤제 등의 형태로 제제화할 수 있다. 비강 흡입제의 경우 디클로로플루오로메탄, 트리클로로플루오로메탄, 디클로로테트라플루오로에탄, 이산화탄소 등의 적합한 추진제를 사용하여 에어로졸 스프레이 형태로 제제화할 수 있으며, 좌제로 제제화할 경우 그 담체로는 위텝솔(witepsol), 트윈(tween) 61, 폴리에틸렌글리콜류, 카카오지, 라우린지, 폴리옥시에틸렌 소르비탄 지방산 에스테르류, 폴리옥시에틸렌 스테아레이트류, 소르비탄 지방산 에스테르류 등을 사용할 수 있다.When the pharmaceutical composition of the present invention is prepared as a parenteral formulation, it may be formulated in the form of an injection, transdermal administration, nasal inhalation, and suppository along with a suitable carrier according to a method known in the art. When formulated as an injection, an aqueous isotonic solution or suspension may be used as a suitable carrier, and specifically, an isotonic solution such as phosphate buffered saline (PBS) containing triethanolamine, sterile water for injection, or 5% dextrose may be used. . When formulated as a transdermal formulation, it may be formulated in the form of ointments, creams, lotions, gels, external solutions, pastas, liniments, air rolls, and the like. In the case of nasal inhalation, it can be formulated in the form of an aerosol spray using a suitable propellant such as dichlorofluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide, etc., and when formulated as a suppository, the carrier is Witepsol ( witepsol), tween 61, polyethylene glycols, cacao fat, laurin fat, polyoxyethylene sorbitan fatty acid esters, polyoxyethylene stearates, sorbitan fatty acid esters, and the like can be used.
약제학적 조성물의 구체적인 제제화와 관련하여서는 당업계에 공지되어 있으며, 예컨대 문헌[Remington's Pharmaceutical Sciences(19th ed., 1995)] 등을 참조할 수 있다. 상기 문헌은 본 명세서의 일부로서 간주 된다.Regarding the specific formulation of the pharmaceutical composition, it is known in the art, and for example, reference may be made to Remington's Pharmaceutical Sciences (19th ed., 1995) and the like. These documents are considered as part of this specification.
본 발명의 약제학적 조성물의 바람직한 투여량은 환자의 상태, 체중, 성별, 연령, 환자의 중증도, 투여 경로에 따라 1일 0.001mg/kg ~ 10g/kg 범위, 바람직하게는 0.001mg/kg ~ 1g/kg 범위일 수 있다. 투여는 1일 1회 또는 수회로 나누어 이루어질 수 있다. 이러한 투여량은 어떠한 측면으로든 본 발명의 범위를 제한하는 것으로 해석되어서는 아니 된다. The preferred dosage of the pharmaceutical composition of the present invention is in the range of 0.001 mg/kg to 10 g/kg per day, preferably 0.001 mg/kg to 1 g, depending on the patient's condition, weight, sex, age, severity of the patient, and route of administration. /kg range. Administration can be done once a day or divided into several times. These dosages should not be construed as limiting the scope of the present invention in any respect.
전술한 바와 같이, 본 발명에 따르면 잔사틴을 이용한 피부 미백용 조성물을 제공할 수 있다.As described above, according to the present invention, a composition for skin whitening using xanthine can be provided.
본 발명의 피부 미백용 조성물은 식품, 화장품, 약품 등으로 제품화될 수 있다.The composition for skin whitening of the present invention can be commercialized as food, cosmetics, medicine, and the like.
도 1은 잔사틴이 마우스 유래 멜라노마 세포주인 B16F10에서 농도 의존적으로 멜라닌 생성을 억제하는 것을 보여주는 결과이다.
도 2는 잔사틴이 마우스 유래 멜라노마 세포주인 B16F10에서 티로시나아제, TRP-1 및 TRP-2 단백질의 발현을 억제함으로 보여주는 웨스턴 블럿 결과이다.1 is a result showing that zansatin inhibits melanin production in a concentration-dependent manner in B16F10, a mouse-derived melanoma cell line.
Figure 2 is a Western blot result showing that zansatin inhibits the expression of tyrosinase, TRP-1 and TRP-2 proteins in B16F10, a mouse-derived melanoma cell line.
이하 본 발명을 실시예 및 실험예를 참조하여 설명한다. 그러나 본 발명의 범위가 이러한 실시예 및 실험예에 한정되는 것은 아니다.Hereinafter, the present invention will be described with reference to Examples and Experimental Examples. However, the scope of the present invention is not limited to these Examples and Experimental Examples.
<< 실시예Example > 잔사틴> Xanxatin 의of 미백 활성 whitening active
1. 재료 및 실험방법1. Materials and test methods
1.1 잔사틴의 준비1.1 Preparation of Xanthine
잔사틴은 자체적으로 천연물에서 분리·동정하여 얻은 것은 사용하였다.Xanthine was independently obtained by isolation and identification from natural products.
1.2 세포배양 1.2 Cell culture
마우스 유래 멜라노마 세포주인 B16F10 cell을 한국세포주은행(Korean Cell Line Bank)으로부터 구입하였으며, 1% 항생제(Gibco, USA)과 10%(100 mg/mL) 우태아혈청(fetal bovine serum)(Gibco, Grand Island, USA)이 함유된 DMEM 배지(Dulbecco's modified Eagle's medium)를 사용하여 37℃, 5% CO2 배양기에서 배양하였으며, 4 일에 한 번씩 계대 배양을 실시하였다.B16F10 cell, a mouse-derived melanoma cell line, was purchased from the Korean Cell Line Bank, and was supplemented with 1% antibiotics (Gibco, USA) and 10% (100 mg/mL) fetal bovine serum (Gibco, USA). Grand Island, USA) containing DMEM medium (Dulbecco's modified Eagle's medium) was used and cultured in a 37°C, 5% CO 2 incubator, and subculture was performed once every 4 days.
1.3 멜라닌 합성량 측정 (Melanin contents) 1.3 Measurement of melanin synthesis (Melanin contents)
B16F10 세포의 배양은 10% 우태아혈청과 100 nM α-MSH(melanocyte stimulating hormone)(Sigma-Aldrich Co.)이 포함된 DMEM 배지를 사용하여 37℃, 5% CO2의 환경에서 세포주를 배양하여 사용하였다. B16F10 cell culture was performed using DMEM medium containing 10% fetal bovine serum and 100 nM α-MSH (melanocyte stimulating hormone) (Sigma-Aldrich Co.) at 37°C and 5% CO 2 by culturing the cell line. used
DMEM 배지로 배양된 B16F10 세포를 24 웰 플레이트에 1 × 105 세포/웰이 되게 분주하고 하루 동안 배양하여 실험에 이용하였다. 이후 시료를 농도별로 처리하고 3일 동안 배양하였다. 배양 후, 인산완충액(pH 7.4)으로 세척하고 1N NaOH를 200㎕씩 각 well에 처리하여 세포를 용해시켰다. 용해된 세포를 96웰 플레이트로 옮기고 마이크로플레이트 리더기(Bioteck, cytation3)를 이용하여 405nm에서 흡광도를 측정하였다. 멜라닌 생성 억제율은 시료 무처리군(α-MSH만의 처리군)인 대조군과 대비해 백분율로 산출하였다.B16F10 cells cultured in DMEM medium were dispensed in a 24-well plate at 1 × 10 5 cells/well and cultured for one day to be used in experiments. Afterwards, the samples were treated by concentration and cultured for 3 days. After culturing, the cells were washed with phosphate buffer (pH 7.4) and treated with 200 μl of 1N NaOH in each well to dissolve the cells. The lysed cells were transferred to a 96-well plate and absorbance was measured at 405 nm using a microplate reader (Bioteck, cytation3). The melanin production inhibition rate was calculated as a percentage compared to the control group, which is a sample untreated group (α-MSH only treated group).
1.4 웨스턴 블럿을 통한 단백질 발현 분석1.4 Analysis of protein expression by Western blot
멜라닌 생성에 관여하는 유전자 중 티로시나아제, TRP-1의 발현 정도를 확인하기 위하여 B16F10 세포를 배양하여 시료를 농도별로 72시간 처리한 후, 상기 세포를 단백질분해효소 저해제(protease inhibitor cocktail, Sigma, st. Louis, MO USA)를 첨가한 RIPA 완충용액(10 mM Tris-HCl(pH 7.5), 1 % NP-40, 0.1 % sodium deoxycholate, 0.1 % SDS, 150 mM NaCl, 1 mM EDTA)으로 용해시켰다. 이후, 얻어진 세포 용해물을 4 ℃에서 5 분간 12,000×g으로 원심분리한 후, BCA 단백질 키트(Thermo Scientific)를 이용하여 단백질의 농도를 측정하였다. 세포에서 추출한 단백질에 전기영동용 완충용액(sample buffer)을 첨가하고 100℃에서 5 분간 가열한 후 SDS-PAGE(sodium dodecyl sulfatepolyacrylamide gel electrophoresis)를 수행하였다. 전기영동 후, 분자량별로 분리된 단백질은 젤에서 니트로셀룰로스 막(nitrocellulose membrane)에 전이시키고 니트로셀룰로스 막에서 정량하였다. 이때, 비특이적 반응을 최소화하기 위해 완충용액(phosphate buffered saline; PBS(pH 7.4), 0.1 % Tween 20, 5 % skimmed milk)을 1 시간 동안 상온에서 반응시킨 후, 1차 항체를 다음과 같이 희석하여 사용하였다. 항체 희석은 5 % 탈지우유 (not-fat dry milk) 용액을 이용하여 상온에서 1 시간 동안 반응시켜 희석하였으며, 희석 비율은 anti-tyrosinase 항체 1:1000, anti-TRP-1 항체 1:1000 하였다. 반응 후, 세척 완충액(0.1 % Tween-20, PBS)으로 5 분간 3 ~ 4회 세척한 후 1:5000으로 희석시킨 2차 항체(HRP-conjugated anti-goat IgG antibody)용액으로 1 시간 동안 상온에서 반응시켰다. 2차 항체 반응 후, ECL(enhanced chemiluminescence) 시약을 사용하여 분석대상 단백질 밴드를 발색시켜 정량하였다. In order to confirm the expression level of tyrosinase and TRP-1 among genes involved in melanin production, B16F10 cells were cultured and samples were treated for 72 hours at each concentration, and then the cells were treated with protease inhibitor cocktail (Sigma, St. Louis, MO USA) was added to the RIPA buffer (10 mM Tris-HCl (pH 7.5), 1% NP-40, 0.1% sodium deoxycholate, 0.1% SDS, 150 mM NaCl, 1 mM EDTA). . Thereafter, the obtained cell lysate was centrifuged at 12,000×g for 5 minutes at 4° C., and the protein concentration was measured using a BCA protein kit (Thermo Scientific). SDS-PAGE (sodium dodecyl sulfatepolyacrylamide gel electrophoresis) was performed after adding a buffer solution for electrophoresis (sample buffer) to the protein extracted from the cells and heating at 100 ° C. for 5 minutes. After electrophoresis, the proteins separated by molecular weight were transferred from the gel to a nitrocellulose membrane and quantified on the nitrocellulose membrane. At this time, in order to minimize non-specific reactions, after reacting in a buffer solution (phosphate buffered saline; PBS (pH 7.4), 0.1
2. 실험 결과2. Experimental results
2.1 멜라닌 합성량 측정(Melanin contents)2.1 Measurement of melanin synthesis (Melanin contents)
결과를 도 1에 나타내었다. 도 1을 참조하여 보면 잔사틴은 양성대조군인 α-MSH 처리군과 비교할 때 농도 의존적으로 멜라닌 생성을 저해하였다. 도 1에는 세포독성 실험 결과도 함께 나타나 있는데, 잔사틴은 5μM까지도 특별한 세포독성을 나타내지 않았다.The results are shown in Figure 1. Referring to FIG. 1, xanthine inhibited melanin production in a concentration-dependent manner when compared to the α-MSH treated group, which is a positive control group. 1 also shows the results of the cytotoxicity test, and xanthine did not show any particular cytotoxicity even at 5 μM.
2.2 웨스턴 블럿 결과2.2 Western blot results
결과를 도 2에 나타내었다. 도 2에서와 확인되는 바와 같이, 시료 처리 농도가 증가할수록 양성대조군인 α-MSH 처리군과 비교할 때 티로시나아제, TRP-1 및 TRP-2 단백질의 발현량이 감소됨을 확인하였다. 따라서 잔사틴은 멜라닌 합성에 관여하는 티로시나아제, TRP-1 및 TRP-2 단백질의 발현을 조절함으로써 멜라닌 합성을 저해하는 것으로 사료 된다. Results are shown in FIG. 2 . As confirmed in FIG. 2, it was confirmed that the expression levels of tyrosinase, TRP-1 and TRP-2 proteins decreased as the sample treatment concentration increased, compared to the α-MSH treatment group, which is a positive control group. Therefore, it is thought that xanthine inhibits melanin synthesis by regulating the expression of tyrosinase, TRP-1 and TRP-2 proteins involved in melanin synthesis.
Claims (4)
A composition for skin whitening comprising xanthine as an active ingredient.
상기 조성물은 화장료 조성물인 것을 특징으로 하는 피부 미백용 조성물.
According to claim 1,
The composition is a composition for skin whitening, characterized in that the cosmetic composition.
상기 조성물은 식품 조성물인 것을 특징으로 하는 피부 미백용 조성물.
According to claim 1,
The composition is a composition for skin whitening, characterized in that the food composition.
상기 조성물은 약제학적 조성물인 것을 특징으로 하는 피부 미백용 조성물.
According to claim 1,
The composition is a composition for skin whitening, characterized in that the pharmaceutical composition.
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