KR20160013226A - Composition for skin moisturizing and alleviating skin inflammation comprising chrysin - Google Patents
Composition for skin moisturizing and alleviating skin inflammation comprising chrysin Download PDFInfo
- Publication number
- KR20160013226A KR20160013226A KR1020160004140A KR20160004140A KR20160013226A KR 20160013226 A KR20160013226 A KR 20160013226A KR 1020160004140 A KR1020160004140 A KR 1020160004140A KR 20160004140 A KR20160004140 A KR 20160004140A KR 20160013226 A KR20160013226 A KR 20160013226A
- Authority
- KR
- South Korea
- Prior art keywords
- skin
- chrysin
- present
- composition
- pharmaceutically acceptable
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
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- 235000015838 chrysin Nutrition 0.000 title claims abstract description 60
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- 229940043370 chrysin Drugs 0.000 title claims abstract description 57
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- A—HUMAN NECESSITIES
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- 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
- A61K8/498—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing heterocyclic compounds with oxygen as the only hetero atom having 6-membered rings or their condensed derivatives, e.g. coumarin
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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
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
- A61K31/35—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom
- A61K31/352—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom condensed with carbocyclic rings, e.g. methantheline
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
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- 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
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- A—HUMAN NECESSITIES
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- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
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- A—HUMAN NECESSITIES
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- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
- A61Q19/007—Preparations for dry skin
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- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
본 발명은 크리신 또는 이의 약학적으로 허용 가능한 염을 포함하는 피부 각질 세포 분화 촉진, 피부 보습 증진, 피부 장벽 강화, 및/또는 피부 염증 완화용 조성물, 특히 화장료 또는 약학 조성물을 제공한다. 본 발명의 화장료 또는 약학 조성물은 부작용이 적어 인체에 안전하면서도 우수한 피부 각질세포 분화능력, 피부 장벽 강화 능력, 피부 보습 증진효과, 및 피부 염증 완화 효과를 가진다.The present invention provides a composition, particularly a cosmetic or pharmaceutical composition, for promoting differentiation of keratinocytes, skin moisturizing enhancement, skin barrier enhancement, and / or skin irritation mitigating action, comprising chrysin or a pharmaceutically acceptable salt thereof. The cosmetic or pharmaceutical composition of the present invention is safe for human body due to its low side effects and has excellent skin keratinocyte differentiation ability, skin barrier enhancement ability, skin moisturizing effect, and skin irritation mitigation effect.
Description
본 발명은 각질세포의 분화 촉진, 피부 보습 증진, 피부 장벽 회복, 피부 염증 완화와 관련된 조성물, 특히 화장료 또는 약학 조성물에 관한 것으로, 인체에 안전하면서도 각질세포 분화 효과, 피부 보습 증진 효과, 피부 장벽 회복 효과, 피부 염증 완화 효과가 우수한 화장료 또는 약학 조성물에 관한 것이다.The present invention relates to a composition, particularly a cosmetic or pharmaceutical composition, related to promotion of differentiation of keratinocytes, enhancement of skin moisturization, restoration of skin barrier and alleviation of skin irritation, and more particularly to a cosmetic composition which is safe for human body, And an effect of alleviating skin irritation.
비타민 D(vitamin D)는 골격 성장 및 유지, 칼슘과 인의 항상성 유지에 필수적이며, 골다공증의 예방과 치료뿐만 아니라 골격계 이외의 조직에서도 중요한 역할을 한다. Vitamin D (vitamin D) is essential for skeletal growth and maintenance, homeostasis of calcium and phosphorus, and plays an important role in the prevention and treatment of osteoporosis as well as in tissues other than the skeletal system.
비타민 D가 세포에 작용하는 조절 기전은 2가지가 있다. 먼저, 비타민 D가 비타민 D 수용체(vitamin D receptor, VDR)에 결합하여 작용하는 것이다. 비타민 D 수용체는 각질세포를 비롯한 여러 세포에서 발현된다. 비타민 D와 비타민 D 수용체의 결합체가 세포 내 핵으로 들어가 비타민 D 인식 부위(vitamin D response elements, VDRE)라 불리는 특정 DNA 염기서열에 결합하여 전사 인자(transcription factor)로 작용하여 표적 유전자(target genes)의 발현을 변화시키는 것이다. 이는 주로 세포 증식에 영향을 주는 유전자의 전사를 일으킨다. 또 다른 하나는 세포 내의 칼슘 이온이 유입되게 하여 작용하는 것이다. 비타민 D는 세포 내의 칼슘 이온의 농도를 증가시키는 일련의 과정에 작용하여, 각질세포의 분화를 촉진시킨다. There are two mechanisms by which vitamin D acts on cells. First, vitamin D binds to the vitamin D receptor (VDR). Vitamin D receptors are expressed in various cells including keratinocytes. The combination of vitamin D and vitamin D receptors enters the intracellular nucleus and binds to a specific DNA sequence called the vitamin D recognition element (VDRE), which acts as a transcription factor, Lt; / RTI > This causes transcription of genes that primarily affect cell proliferation. The other is to allow the intracellular calcium ions to flow. Vitamin D acts on a series of processes that increase the concentration of calcium ions in the cell, thereby promoting the differentiation of keratinocytes.
이런 두 가지 기전의 조합이 표적 세포의 증식을 억제하고, 분화를 촉진하게 만든다. 세포 증식이나 세포 분화의 역할 이외에도 비타민 D는 면역 조절 기능을 한다. 인터루킨(IL)-1에 대한 T 세포의 반응을 억제하고, 면역 반응에 중요한 인터루킨-2, 인터루킨-6, 인터페론-γ생산도 억제한다. 또한, T 세포 활성 억제를 증진시키며, NK (natural killer) 세포의 형성도 억제한다. 이러한 비타민 D의 세포 증식 감소, 세포 분화 촉진, 면역 조절 기능 등의 조합이 비타민 D와 그 유도체들이 건선 치료에 효과를 보이는 이유를 설명할 수 있다. The combination of these two mechanisms inhibits the proliferation of target cells and promotes differentiation. In addition to cell proliferation and cell differentiation, vitamin D also has immunomodulatory functions. It inhibits the T cell response to IL-1 and inhibits interleukin-2, interleukin-6, and interferon-γ production, which are important for the immune response. In addition, it promotes inhibition of T cell activation and inhibits the formation of NK (natural killer) cells. These combinations of vitamin D, cell proliferation, cell differentiation, and immunomodulation can explain why Vitamin D and its derivatives are effective in the treatment of psoriasis.
그러나, 이러한 비타민 D와 비타민 D의 유도체는 화장품에 사용할 수 없는 금지 원료이고, 화장품에 사용할 수 있는, 비타민 D 수용체의 전사 활성을 증가시킬 수 있는 물질에 대한 연구는 아직 이루어지고 있지 않다. However, these vitamin D and vitamin D derivatives are prohibited raw materials that can not be used in cosmetics, and studies on substances capable of increasing the transcriptional activity of vitamin D receptors, which can be used in cosmetics, have not yet been conducted.
따라서, 상기와 같은 단점 및 부작용은 해소하면서, 화장품 등에 안정적으로 사용할 수 있는 각질세포의 분화 촉진, 피부 보습 증진, 피부 장벽 회복, 피부 염증 완화와 관련된 효능을 갖는 유용한 활성 성분에 대한 요구가 꾸준히 있어 왔다.Therefore, there is a demand for a useful active ingredient having efficacy related to promotion of keratinocyte differentiation, skin moisturization, skin barrier recovery, and skin irritation relieving which can be stably used in cosmetics and the like while solving the above-mentioned disadvantages and side effects come.
따라서 본 발명은 위와 같은 문제점을 해결하고, 부작용이 적고 인체에 안전하면서도 각질세포의 분화 촉진, 피부 보습 증진, 피부 장벽 회복, 및 피부 염증 완화 효과가 우수한 새로운 활성 성분, 즉, 활성 성분의 이러한 유용한 용도를 제공하는 것이다.Accordingly, it is an object of the present invention to provide a new active ingredient which is excellent in the effect of promoting the differentiation of keratinocytes, promoting skin moisturization, restoring skin barrier and alleviating skin inflammation, that is, It is intended to provide use.
다시 말해, 본 발명이 해결하고자 하는 과제는 상기 효능이 우수한 활성 성분을 유효성분으로 포함하는 조성물, 특히 화장료 또는 약학 조성물을 제공하고자 한다.In other words, a problem to be solved by the present invention is to provide a composition, particularly a cosmetic composition or a pharmaceutical composition, containing the active ingredient with excellent efficacy as an active ingredient.
상기 과제를 해결하기 위하여, 본 발명은 크리신 또는 이의 약학적으로 허용 가능한 염을 유효성분으로 포함하는 각질세포의 분화 촉진용, 피부 보습 증진용, 피부 장벽 회복용, 또는 피부 염증 완화용 조성물, 바람직하게는 화장료 조성물 또는 약학 조성물을 제공한다. The present invention provides a composition for accelerating differentiation of keratinocyte, which comprises chrysin or a pharmaceutically acceptable salt thereof as an active ingredient, for skin moisturizing enhancement, for restoring skin barrier, or for alleviating skin irritation, Preferably a cosmetic composition or a pharmaceutical composition.
즉, 본 발명은 크리신 또는 이의 약학적으로 허용 가능한 염의 각질세포의 분화 촉진용, 피부 보습 증진용, 피부 장벽 회복용, 또는 피부 염증 완화용이라는 새로운 용도를 제공한다.That is, the present invention provides a novel use of chrysin or a pharmaceutically acceptable salt thereof for promoting differentiation of keratinocytes, for skin moisturizing, for restoring skin barrier, or for alleviating skin irritation.
더 나아가 본 발명은 피부에 도포했을 때 피부 각질 형성세포(keratinocyte)의 분화(differentiation)를 정상적으로 환원 및 유지시킴으로써, 표피분화가 불완전함으로 해서 발생하는 피부건조 증상, 아토피 증상, 건선 및 접촉성 피부염 증상 등을 예방 및/또는 개선하는 효과가 있는 조성물을 제공할 수 있다. Further, the present invention provides a method for treating skin diseases, such as dry skin symptoms, atopic symptoms, psoriasis, and contact dermatitis symptoms, which are caused by incomplete epidermal differentiation by normally reducing and maintaining differentiation of keratinocytes when applied to skin And the like can be provided.
그리고 본 발명의 조성물은 피부 장벽을 강화시켜 피부의 수분보유능력을 증진시키고, 피부의 유연성과 견고성을 높여 피부가 본래의 보호기능을 원활히 수행하게 하며, 피부건조증을 개선할 뿐만 아니라 피부건조로 인한 잔주름의 발생도 감소시킬 수 있다. In addition, the composition of the present invention enhances the water retention ability of the skin by enhancing the skin barrier and improves the flexibility and durability of the skin to smoothly perform the original protective function of the skin, The occurrence of fine wrinkles can be reduced.
또한, 본 발명은 비타민 D 수용체의 전사를 활성화시킴으로써, 비타민 D와 유사한 효과를 가지면서도 안전성이 향상된 비타민 D 수용체 전사 활성화용 조성물, 특히 화장료 또는 약학 조성물을 제공한다.The present invention also provides a composition for activating transcription of vitamin D receptors, particularly a cosmetic or pharmaceutical composition, which has an effect similar to that of vitamin D and has improved safety by activating transcription of vitamin D receptor.
본 발명의 크리신은 화학식은 C15H10O4 이고 분자량은 254.24로서, 아래 화학식 1로 나타낼 수 있다.The chrysin of the present invention has a chemical formula of C 15 H 10 O 4 and a molecular weight of 254.24, which can be represented by the following
[화학식 1][Chemical Formula 1]
상기 화합물은 시계초류(Passiflora caerulea, Passiflora incarnate 등), 캐모마일, 오록실룸 인디쿰(Oroxylum indicum) 등을 포함한 식물에서 분리할 수 있으나, 본 발명은 이러한 크리신의 획득 방법에 한정되지 않는다. 본 발명에 따른 크리신은 당 업계에 공지된 방법으로 화학적으로 합성하거나, 시판되는 물질을 사용할 수 있다.The compound was identified as Passiflora caerulea , Passiflora incarnate, etc.), chamomile, orroyl silk indium ( Oroxylum indicum ) and the like, but the present invention is not limited to the method for obtaining such a chrysin. The chrysin according to the present invention may be chemically synthesized by a method known in the art, or a commercially available material may be used.
본 발명의 "약학적으로 허용 가능한 염"은 독성이 없거나 적은 산 또는 염기로 제조된 염들을 말한다. 본 발명의 화합물이 상대적으로 산성일 경우 염기(base) 부가 염들은 충분한 양의 원하는 염기와 적당한 비활성(inert) 용매로 그러한 화합물의 중성 형태를 접촉하여 얻을 수 있다. 약학적으로 허용 가능한 염기 부가 염은 나트륨, 칼륨, 칼슘, 암모늄, 마그네슘 또는 유기 아미노로 이루어진 염을 포함하나, 이에 한정되는 것은 아니다. 본 발명의 화합물이 상대적으로 염기성일 경우 산(acid) 부가 염들은 충분한 양의 원하는 산과 적당한 비활성 용매로 그러한 화합물들의 중성 형태를 접촉하여 얻을 수 있다. 약학적으로 허용 가능한 산 부가 염은 프로피온산, 이소부틸산, 옥살산, 사과산, 말론산, 안식향산, 호박산, 수버릭(suberic), 푸마르산, 만데릭산, 프탈릭산, 벤젠설폰산, p-토릴설폰산, 구연산, 주석산, 메탄설폰산, 염산, 브롬산, 질산, 탄산, 일수소탄산(monohydrogencarbonic), 인산, 일수소인산, 이수소인산, 황산, 일수소황산, 요오드화수소, 아인산(phosphorous acid) 등으로 형성된 염을 포함하나, 이에 한정되는 것은 아니다. 또한 알지네이트(arginate) 같은 아미노산의 염 및 글루쿠로닉(glucuronic) 또는 갈락투노릭(galactunoric) 산들과 같은 유기산의 유사체를 포함하나, 이에 한정되는 것은 아니다.The term "pharmaceutically acceptable salts " of the present invention refers to salts prepared with little or no toxicity or with acids or bases. When the compound of the present invention is relatively acidic, the base addition salts may be obtained by contacting a neutral form of such compound with a sufficient amount of the desired base and an appropriate inert solvent. Pharmaceutically acceptable base addition salts include, but are not limited to, salts formed with sodium, potassium, calcium, ammonium, magnesium or organic amino. Where the compounds of the present invention are relatively basic, acid addition salts can be obtained by contacting neutral forms of such compounds with a sufficient amount of the desired acid and a suitable inert solvent. Pharmaceutically acceptable acid addition salts include those derived from organic acids such as propionic acid, isobutyric acid, oxalic acid, malic acid, malonic acid, benzoic acid, succinic acid, sueric, fumaric acid, mandelic acid, phthalic acid, benzenesulfonic acid, Phosphoric acid, monohydrogencarbonic acid, phosphoric acid, monohydrogenphosphoric acid, dihydrogenphosphoric acid, sulfuric acid, monohydrogensulfuric acid, hydrogen iodide, phosphorous acid, And the like, but are not limited thereto. But are not limited to, salts of amino acids such as arginate and analogs of organic acids such as glucuronic or galactunoric acids.
본 발명의 크리신은 수화물, 에탄올화물 등의 형태를 포함하는 용매화된 형태뿐만 아니라 비-용매화된(unsolvated) 형태로 존재할 수도 있다. 본 발명의 크리신은 결정형 또는 무정형 형태로 존재할 수 있으며, 이러한 모든 물리적 형태는 본 발명의 범위에 포함된다.The chrysins of the present invention may exist in solvated as well as unsolvated forms, including forms such as hydrates, ethanolates, and the like. The chrysins of the present invention may exist in crystalline or amorphous form, and all such physical forms are included within the scope of the present invention.
본 발명의 크리신은 또한 전구약물(pro-drug)의 형태로 존재할 수 있다. 특히, 피부에 존재하는 에스테라제 또는 아미다제에 의해 활성 형태인 크리신으로 분리될 수 있는 전구약물이 사용될 수 있다. 구체적으로, 친수성/친유성, 피부흡수성, 용해도, 크리신의 체내 동태 등을 조절하기 위하여 탄소수 1~6의 알킬, 탄소수 1~6의 알케닐, 탄소수 1~6의 할로알킬기, 탄소수 1~6의 히드록시알킬기, 알콕시 및 알킬부위가 각각 탄소수 1~6인 알콕시알킬 또는 알콕시알콕시알킬기, 펜아실기, 탄소수 1~6의 알킬로 치환된 펜아실기, 알콕시 및 알킬부위가 각각 탄소수 1~6인 알콕시카르보닐알킬기, 탄소수 1~6의 시아노알킬기, 각각의 알킬부위가 각각 탄소수 1~6인 알킬티오알킬기 등의 치환기가 에스테르 결합 또는 아마이드 결합에 의해 크리신의 -OH 기에 하나 이상 연결될 수 있다. 이러한 전구약물은 BURGER'S MEDICINAL CHEMISTRY AND DRUG DISCOVERY, 172-178, 949-982 (Manfred E. Wolffed., 5th ed. 1995) 등에 기재된 방법과 같은 본 발명이 속한 분야에서 잘 알려진 방법에 의해 만들어질 수 있다.The chrysins of the present invention may also be present in the form of pro-drugs. In particular, prodrugs that can be separated into the active form chrysocin by esterases or amidases present in the skin may be used. Specifically, in order to control hydrophilicity / lipophilicity, skin absorbency, solubility and kinetics of the body of krypticine, alkyl having 1 to 6 carbon atoms, alkenyl having 1 to 6 carbon atoms, haloalkyl group having 1 to 6 carbon atoms, An alkoxyalkyl group having 1 to 6 carbon atoms, an alkoxyalkyl or alkoxyalkoxyalkyl group having 1 to 6 carbon atoms, a phenacyl group, a phenacyl group substituted with an alkyl having 1 to 6 carbon atoms, an alkoxy group having 1 to 6 carbon atoms, A cyanoalkyl group having 1 to 6 carbon atoms, an alkylthioalkyl group having 1 to 6 carbon atoms in each alkyl moiety, etc., may be linked to the -OH group of the chrysin by an ester bond or an amide bond. Such prodrugs can be made by methods well known in the art, such as those described in BURGER'S MEDICAL CHEMISTRY AND DRUG DISCOVERY, 172-178, 949-982 (Manfred E. Wolffed., 5th ed. .
본 발명의 발명자들은 여러 가지 성분들을 대상으로 연구를 거듭한 결과, 크리신 또는 이의 약학적으로 허용 가능한 염이 각질세포 분화 촉진효과 및 피부 장벽 강화 효과뿐만 아니라, 우수한 피부 보습 증진 능력 및 피부 염증 완화 능력을 나타낸다는 것을 발견하게 되었다.The inventors of the present invention have conducted research on various components and found that chrysin or a pharmaceutically acceptable salt thereof not only has a promoting effect of keratinocyte differentiation and a skin barrier strengthening effect but also an excellent skin moisturizing ability and a skin inflammation- I was able to discover that it represents ability.
따라서, 본 발명은 각질세포 분화를 촉진하고, 피부 장벽 강화 효과를 나타내며, 우수한 피부 보습 증진 능력 및/또는 피부 염증 완화 능력을 갖는 화장료 조성물을 제공할 수 있다. Accordingly, the present invention can provide a cosmetic composition that promotes keratinocyte differentiation, exhibits a skin barrier strengthening effect, has excellent skin moisturizing ability and / or skin irritation alleviating ability.
또한, 본 발명은 크리신을 유효성분으로 포함하는 각질세포 분화 촉진용, 피부 장벽 강화용, 피부 보습용, 및/또는 피부 염증 완화용 약학 조성물을 제공할 수 있다. In addition, the present invention can provide a pharmaceutical composition for promoting differentiation of keratinocytes, enhancing skin barrier, moisturizing skin, and / or alleviating skin inflammation comprising chrysin as an effective ingredient.
본 발명의 조성물은 각질세포 분화 촉진, 피부 장벽 강화, 피부 보습, 및/또는 피부 염증 완화를 위한 상기 크리신 이외에 본 발명의 효과를 해치지 않는 범위 내에서 보습제, 증점제, 계면활성제, 유상기제, 방부제, 산화방지제, 알코올, 향료, pH 조절제, 천연추출물 등은 물론 본 발명의 효과를 배가시키기 위한 화학적 및/또는 천연 성분을 적당한 양으로 배합할 수 있다.The composition of the present invention may contain, in addition to the above-mentioned chrysin for promoting keratinocyte differentiation, skin barrier enhancement, skin moisturizing, and / or skin irritation mitigating effect of the present invention, humectants, thickeners, surfactants, , An antioxidant, an alcohol, a flavor, a pH adjuster, a natural extract, and the like, as well as chemical and / or natural ingredients for enhancing the effects of the present invention.
본 발명의 조성물은 의약품, 기능성 식품 및 화장품의 형태로 제조될 수 있다. 이러한 의약품, 기능성 식품 및 화장품은 약제학적으로 또는 화장학적으로 허용되는 부형제 또는 첨가제를 포함할 수 있다. 본 발명의 조성물은 단독으로 혹은 어떤 편리한 운반체, 부형제 등과 함께 혼합하여 투여될 수 있고, 그러한 투여 제형은 단회투여 또는 반복투여 제형일 수 있다. The composition of the present invention can be manufactured in the form of medicines, functional foods and cosmetics. Such medicaments, functional foods and cosmetics may include pharmaceutically or cosmetically acceptable excipients or additives. The compositions of the present invention may be administered alone or in admixture with any convenient vehicle, excipient, etc., and such dosage forms may be single-dose or multiple-dose formulations.
본 발명의 조성물은 고형 제제, 반고형제제 또는 액상 제제일 수 있다. 고형 제제는 산제, 과립제, 정제, 캅셀제, 좌제 등이 있으나, 이에 한정되는 것은 아니다. 고형 제제에는 부형제, 착향제, 결합제, 방부제, 붕해제, 활택제, 충진제 등이 포함될 수 있으나 이에 한정되는 것은 아니다. 반고형 제제로는 크림제, 로션제, 유화제, 리니멘트제 등이 있으나, 이에 한정되는 것은 아니며, 적당한 착색제, 착향제, 안정화제, 점성화제, 계면활성제 등을 첨가하여 제조할 수 있다. 액상 제제로는 물, 알코올, 프로필렌 글리콜 용액 같은 용액제, 현탁액제, 유제 등이 있으나, 이에 한정되는 것은 아니며, 적당한 착색제, 착향제, 안정화제, 점성화제 등을 첨가하여 제조할 수 있다.The composition of the present invention may be a solid preparation, a semi-solid preparation or a liquid preparation. Solid preparations include, but are not limited to, powders, granules, tablets, capsules, suppositories, and the like. Solid form preparations may include, but are not limited to, excipients, flavoring agents, binders, preservatives, disintegrants, lubricants, fillers, and the like. Examples of the semi-solid preparation include a cream, a lotion, an emulsifier, a liniment, and the like, but the present invention is not limited thereto, and can be produced by adding a suitable colorant, a flavoring agent, a stabilizer, a tackifier, a surfactant, and the like. Examples of the liquid preparation include water, alcohols, solutions such as solutions of propylene glycol, suspensions, emulsions and the like, but not limited thereto, and they can be prepared by adding appropriate coloring agents, flavoring agents, stabilizers, tackifiers and the like.
예를 들어, 산제는 본 발명의 크리신 또는 이의 염과 유당, 전분, 미결정셀룰로오스 등 약제학적으로 허용되는 적당한 부형제를 단순 혼합함으로써 제조될 수 있다. 과립제는 본 발명의 크리신 또는 이의 염; 약제학적으로 허용되는 적당한 부형제; 및 폴리비닐피롤리돈, 히드록시프로필셀룰로오스 등의 약제학적으로 허용되는 적당한 결합제를 혼합한 후, 물, 에탄올, 이소프로판올 등의 용매를 이용한 습식과립법 또는 압축력을 이용한 건식과립법을 이용하여 제조될 수 있다. 또한 정제는 상기 과립제를 마그네슘스테아레이트 등의 약제학적으로 허용되는 적당한 활택제화 혼합한 후, 타정기를 이용하여 타정함으로써 제조될 수 있다. 또한 예를 들어, 피부외용제는 바세린, 스테아릴알콜 등의 약제학적으로 또는 화장학적으로 허용되는 적당한 기재; 폴리소르베이트, 솔르비탄 세스퀴올레이트 등의 약제학적으로 또는 화장학적으로 허용되는 적당한 계면활성제; 글리세린 등의 약제학적으로 또는 화장학적으로 허용되는 적당한 보습제; 약제학적으로 또는 화장학적으로 허용되는 적당한 용제; 및 착향제, 착색제, 안정화제, 점성화제 등을 균질하게 혼합하는 통상의 피부외용제 제조방법에 의해서 제조될 수 있다.For example, powders can be prepared by simple mixing of the chrysin or salt thereof of the present invention with an appropriate pharmaceutically acceptable excipient such as lactose, starch, microcrystalline cellulose and the like. The granule may be a creosin or a salt thereof of the present invention; Suitable excipients which are pharmaceutically acceptable; And a suitable pharmaceutically acceptable binder such as polyvinylpyrrolidone and hydroxypropylcellulose, followed by wet granulation using a solvent such as water, ethanol or isopropanol or dry granulation using a compressive force . Further, the tablet may be prepared by mixing the above granule with a suitable pharmaceutically acceptable salt such as magnesium stearate and then tableting it using a tablet machine. Also, for example, the external preparation for skin may be a pharmaceutically or cosmetically acceptable suitable substance such as vaseline, stearyl alcohol and the like; A suitable pharmaceutically or cosmetically acceptable surfactant such as polysorbate, sorbitan sesquioleate, and the like; Suitable pharmacologically or cosmetically acceptable moisturizing agents such as glycerin; A suitable pharmaceutically or cosmetically acceptable solvent; And a conventional external preparation for skin preparation in which a flavoring agent, a coloring agent, a stabilizer, a tackifier and the like are homogeneously mixed.
한편, 본 발명의 크리신을 함유하는 화장료 조성물은 화장수류, 로션류, 크림류와 같은 다양한 제형으로 제조될 수 있으며, 구체적으로는 에멀젼상, 크림상, 페이스트상 등 다양한 성상으로 적용이 가능하고, 통상적인 화장료 제조법을 이용하여 제조할 수 있다.On the other hand, the cosmetic composition containing the chrysin of the present invention can be manufactured into various formulations such as cosmetics, lotions, and creams, and specifically, it can be applied in various forms such as emulsion, cream, and paste, Can be produced by using the cosmetic preparation method of the present invention.
본 발명의 화장료 조성물 또는 약학 조성물은 크리신 또는 이의 약학적으로 허용 가능한 염을 조성물 총 중량 대비 0.0005 내지 10 중량% 포함하며, 바람직하게는 0.001 내지 5 중량% 포함한다.The cosmetic composition or the pharmaceutical composition of the present invention contains 0.0005 to 10% by weight, preferably 0.001 to 5% by weight, of chrysin or a pharmaceutically acceptable salt thereof, based on the total weight of the composition.
상기 크리신 또는 이의 염이 0.0005 중량% 미만이면 본래 목적하는 각질세포 분화 촉진 효과, 피부 보습 증진 효과, 피부 장벽 회복 효과 및/또는 피부 염증 완화 효과를 충분하게 달성할 수 없어 바람직하지 못하며, 10 중량%를 초과하면 증가하는 함량만큼 뚜렷한 효과의 증대가 기대되지 않아 비경제적이며, 화장료 및 약학 조성물의 안정성에 문제를 발생시킬 수 있기 때문이다.If the amount of the above-mentioned chrysin or its salt is less than 0.0005% by weight, it is undesirable because it can not sufficiently attain the desired effect of accelerating keratinocyte differentiation, skin moisturizing effect, skin barrier restoration effect and / %, It is unexpected that the increase in the effect is not expected to increase as much as the content, which may cause problems in stability of cosmetic and pharmaceutical compositions.
본 발명에 따른 각질세포 분화 촉진 효과, 피부 보습 증진 효과, 피부 장벽 회복 효과 및/또는 피부 염증 완화 효과라는 목적을 달성하기 위하여 본 발명의 크리신 또는 이의 약학적으로 허용 가능한 염은 매일 약 0.01 mg/kg 내지 약 10 g/kg이 투여될 수 있으며, 약 0.1 mg/kg 내지 약 1 g/kg의 1일 투여 용량이 바람직하다. 그러나 상기 투여량은 투여 경로 및 제형, 환자의 상태(연령, 성별, 체중 등), 치료하고 있는 상태의 심각성 등에 따라 다양할 수 있다. 필요에 따라 편리성을 위하여 1일 총 투여량이 나누어지고 하루 동안 여러 번 나누어 투여될 수 있다.In order to achieve the object of promoting keratinocyte differentiation promotion, skin moisturizing effect, skin barrier restoration effect and / or skin inflammation relieving effect according to the present invention, the chrysin or a pharmaceutically acceptable salt thereof of the present invention is administered in an amount of about 0.01 mg / kg to about 10 g / kg, and a daily dosage of about 0.1 mg / kg to about 1 g / kg is preferred. However, the dosage may vary depending on the route of administration and formulation, the patient's condition (age, sex, weight, etc.), severity of the condition being treated, and the like. For convenience, the total daily dose may be divided as needed and divided into several doses throughout the day.
본 발명의 크리신 또는 이의 약학적으로 허용 가능한 염을 함유하는 조성물은 피부 각질세포 분화를 촉진한다.Compositions containing the chrysin or a pharmaceutically acceptable salt thereof of the present invention promote dermal keratinocyte differentiation.
아울러 피부 각질세포 분화를 촉진함으로써 피부 건조 증상, 아토피 증상, 건선 및 접촉성 피부염 증상을 완화 또는 개선하는 효과를 나타낸다.In addition, it promotes the differentiation of keratinocytes and thus alleviates or alleviates the symptoms of dry skin, atopic symptoms, psoriasis and contact dermatitis.
본 발명의 크리신 또는 이의 약학적으로 허용 가능한 염을 함유하는 조성물은 피부 장벽을 강화시키며, 피부의 유연성 및 견고성을 높이고, 피부의 수분 보유 능력을 증가시키는 효과를 나타낸다.The composition containing the chrysin or a pharmaceutically acceptable salt thereof of the present invention strengthens the skin barrier, increases the flexibility and firmness of the skin, and increases the water retention capacity of the skin.
본 발명의 크리신 또는 이의 약학적으로 허용 가능한 염을 함유하는 조성물은 피부 보습 증가 효과 및 피부 염증 완화 효과를 나타낸다. The composition containing the chrysin or a pharmaceutically acceptable salt thereof of the present invention exhibits a skin moisturizing effect and a skin inflammation relieving effect.
또한, 비타민 D 수용체의 전사를 활성화시켜 각질세포에서 비타민 D와 유사한 효과를 발휘함으로써 안전하면서도 우수한 효과의 화장품 원료로 이용될 수 있다.In addition, by activating transcription of the vitamin D receptor, it exerts a similar effect to vitamin D in keratinocytes, and thus it can be used as a safe and excellent cosmetic material.
도 1은 리포터 유전자 측정법을 이용하여, 크리신의 농도별 VDR 전사 활성을 측정한 결과를 보여주는 그래프이다.
도 2는 quantitative real time PCR을 이용하여, 인체 유래의 피부 각질세포에서 크리신이 VDR의 mRNA 발현에 미치는 영향을 나타낸 그래프이다.
도 3은 quantitative real time PCR을 이용하여, 인체 유래의 피부 각질세포에서 크리신이 Involucrin의 mRNA 발현에 미치는 영향을 나타낸 그래프이다.
도 4는 quantitative real time PCR을 이용하여, 인체 유래의 피부 각질세포에서 크리신이 Fillaggrin의 mRNA 발현에 미치는 영향을 나타낸 그래프이다.FIG. 1 is a graph showing the results of measurement of VDR transcriptional activity by concentration of chrysin using a reporter gene measurement method.
FIG. 2 is a graph showing the effect of chrysin on mRNA expression of VDR in human keratinocytes using quantitative real time PCR.
FIG. 3 is a graph showing the effect of chrysin on mRNA expression of Involucrin in human keratinocytes using quantitative real time PCR.
FIG. 4 is a graph showing the effect of chrysin on mRNA expression of Fillaggrin in human keratinocytes using quantitative real time PCR.
이하, 본 발명을 보다 구체적으로 설명하기 위하여 하기 실시예 등을 들어 설명한다. 그러나, 본 발명에 따른 실시예들은 여러 가지 다른 형태로 변형될 수 있으며 본 발명의 범위가 아래에서 상술하는 실시예들에 한정되는 것으로 해석되어서는 안 된다. 본 발명의 실시예들은 본 발명의 구체적 이해를 돕기 위해 예시적으로 제공되는 것이다.Hereinafter, the present invention will be described in more detail with reference to the following examples. However, the embodiments according to the present invention can be modified into various other forms, and the scope of the present invention should not be construed as being limited to the above-described embodiments. The embodiments of the present invention are provided by way of example to facilitate a specific understanding of the present invention.
본 발명의 하기 실시예에 사용한 크리신은 지안 내츄럴 필드 바이오테크닉(Xian natural field biotechnique Co. Ltd., 중국)에서 구입하여 사용하였다.The chrysin used in the following examples of the present invention was purchased from Xian natural field biotechnology Co. Ltd., China.
<실시예 1> 비타민 D 수용체(VDR) 전사 활성화 평가Example 1 Evaluation of Transcriptional Activation of Vitamin D Receptor (VDR)
인체 각질 세포주(human keratinocyte)인 HaCaT을 스테로이드 호르몬이 제거된 우태아 혈청(5% charcoal stripped FBS)을 포함하는 Dulbecco's modified Eagle's medium (DMEM; Invitrogen, 미국) 배지에 계대 배양하였다. 플라즈미드는 VDRE reporter (Qiagen, 독일)를 구입하여 사용하였는데, 이것은 VDR에 의해 리포터(reporter) 유전자인 반딧불 루시퍼라아제(firefly luciferase)가 발현할 수 있도록 만들어진 플라즈미드와 레닐라 루시퍼라아제(Renilla luciferase)를 항상 발현하는 플라즈미드가 40 : 1 의 비율로 들어있는 DNA 혼합물이다. HaCaT 세포 5x104 개를 24 웰(well) 플레이트에 분주하고 18시간 배양한 후, 상기 플라즈미드 유전자인 VDRE reporter 1 ㎍을 lipofectaminTM reagent(Invitrogen, 미국)를 이용하여 제조사의 설명서에 따라 일시적 트랜스팩션(transient transfection) 하였다. 이 세포들을 24 시간 배양 후, 인산화완충용액(PBS)로 세척한 다음 우태아 혈청이 없는 DMEM 배지에 크리신을 농도별(0.02, 0.2, 2㎍/ml)로 첨가하였다. 음성 대조군으로는 크리신을 녹일 때 사용한 DMSO를 처리한 군을 이용하였다. 24시간 배양 후, 인산완충용액으로 세척하고, 세포 용해 완충액을 이용하여 세포를 용해한 후 원심분리기를 이용하여 상등액만 추출하여 루시퍼라아제 어세이를 실시한 후 퍼킨엘머 빅터3(Perkin Elmer Victor3, 미국)로 발광정도(luminescence)를 측정하고 도 1에 나타내었다.HaCaT, a human keratinocyte, was subcultured in Dulbecco's modified Eagle's medium (DMEM; Invitrogen, USA) containing steroid hormone-free fetal bovine serum (5% charcoal stripped FBS). Plasmids were purchased from VDRE reporter (Qiagen, Germany), which was constructed by plasmids and Renilla luciferase, which was made by VDR to express the reporter gene firefly luciferase, Lt; RTI ID = 0.0 > 40: < / RTI > HaCaT cells, 5x10 4 gae by the use of a 24-well (well) was dispensed to the plate, and 18 hours of incubation, the
도 1에 나타나는 바와 같이, 본 발명에 따른 크리신은 농도 의존적으로 유의한 정도의 VDR의 활성을 증가시켜 높은 루시퍼라아제 활성을 나타냄을 알 수 있다.As shown in FIG. 1, it can be seen that the chrysin according to the present invention increases the activity of VDR to a significant extent in a concentration-dependent manner, indicating high luciferase activity.
<실시예 2> VDR, Involucrin 및 Fillagrin 유전자 발현 조절 효과<Example 2> Modulation effect of VDR, Involucrin and Fillagrin gene expression
인체 유래의 각질 세포(normal human keratinocytes)는 ATCC(미국)에서 구입하여 사용하였다. Dermal basal cell media(DBCM, ATCC, 미국)를 사용하였고, 각질세포 성장인자(keratinocyte growth factor, KGF, ATCC, 미국)를 넣어서 사용하였다. 2x105 개의 세포를 6 웰 플레이트에 분주하여 18시간 배양한 후, 2 ㎍/ml 농도의 크리신을 48시간 동안 처리하였다. 세포를 회수하여 PBS로 세척한 후, RNeasy mini kit (Qiagen, Germany)를 이용하여 전체 RNA를 추출한 뒤, 정량하여 1 ㎍의 RNA를 GeneAmp® RNA PCR kit (Applied Biosystems, USA)를 이용하여 역전사(reverse transcription)하였다. 역전사 반응은 Mycycler® PCR 기기(Biorad, USA)를 이용하여 수행하였다.Human keratinocytes were purchased from ATCC (USA). Dermal basal cell media (DBCM, ATCC, USA) was used and keratinocyte growth factor (KGF, ATCC, USA) was used. 2 x 10 5 cells were plated in 6-well plates and cultured for 18 hours, followed by treatment with chrysin at a concentration of 2 μg / ml for 48 hours. Cells were harvested and washed with PBS. Total RNA was extracted using RNeasy mini kit (Qiagen, Germany), and 1 μg of RNA was quantitatively analyzed using GeneAmp ® RNA PCR kit (Applied Biosystems, USA) reverse transcription). Reverse transcription was performed using a Mycycler ® PCR instrument (Biorad, USA).
합성된 cDNA는 한 반응 당 300ng을 사용하였고, iTaq™ Fast SYBR®Green Supermix With ROX (Biorad, USA)와 유전자 특이적인 프라이머를 사용하여 quantitative real time PCR을 수행하였다. Real time PCR은 CFX96 Touch™ Real-Time PCR Detection System 기기 (Biorad, 미국)를 이용하여 수행하였다. Quantitative real time PCR에 사용된 프라이머 서열은 표 1과 같다. 모든 프라이머는 필코리아(Korea)에서 주문 제작하였다. Real time PCR을 통해 얻은 실험 결과는 housekeeping 유전자인 Glyceraldehyde-3-phosphate dehydrogenase (GAPDH)를 기준으로 ΔΔCt 방법(Livak KJ, Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method, Methods, 25, 402-408(2001)으로 계산하여 나타내었다. 또한, 음성 대조군의 mRNA 발현량을 1.0으로 기준하여 크리신 처리 실험군의 mRNA 발현량을 수치화하여 도 2 내지 4에 나타내었다. Quantitative real time PCR was performed using iTaq ™ Fast SYBR ® Green Supermix With ROX (Biorad, USA) and gene specific primers. Real-time PCR was performed using a CFX96 Touch ™ Real-Time PCR Detection System instrument (Biorad, USA). The primer sequences used in the quantitative real time PCR are shown in Table 1. All primers were custom made in Korea. Experimental results obtained by real-time PCR were compared with those of the housekeeping gene Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) using the ΔΔCt method (Livak KJ, Schmittgen TD: Analysis of relative gene expression data using real- Delta Delta C (T)) Method, Methods, 25, 402-408 (2001). In addition, by quantifying the mRNA expression level of the test group treated with chrysin using the negative control group mRNA expression level as 1.0 2 to 4.
도 2 내지 4에 나타나는 바와 같이, 본 발명에 따른 크리신은, VDR 전사 활동을 유도하거나 발현을 증가시킴으로써, 피부 각질 세포의 분화를 촉진하고 피부 보습을 증가시켰다. 피부 각질 세포의 분화의 중요한 마커(marker)인 인볼루크린(involurin)의 발현을 증가시켜 각질세포의 분화를 촉진하였으며, 피부의 보습에 중요한 천연보습인자(natural moisturizing factor, NMF)의 전구체인 필라그린(Fillaggrin)의 유전자 발현을 증가시키는 효과를 나타냈다. 이를 통해, 본 발명에 따른 크리신 또는 이의 약학적으로 허용 가능한 염은 각질 세포 분화를 촉진하고, 피부 수분 보유능력을 증가하는 것으로 생각된다. 더 나아가 본 발명은 상기 결과로부터 피부의 표피분화가 불완전하여 발생하는 피부 건조 증상, 아토피 증상 등의 예방 또는 개선에 효과가 있을 것으로 생각되며, 피부의 유연성과 견고성을 높이고 피부의 수분보유능력을 증진시켜 피부 보습에 탁월한 효과를 나타낼 것으로 생각된다.As shown in FIGS. 2 to 4, the chrysin according to the present invention promotes differentiation of dermal keratinocytes and increases skin moisturization by inducing VDR transcription activity or increasing expression. It promotes the differentiation of keratinocytes by increasing the expression of involurin, an important marker of keratinocyte differentiation, and promotes the differentiation of keratinocytes, which is a precursor of natural moisturizing factor (NMF) Green (Fillaggrin) gene expression was shown to increase the effect. Accordingly, it is believed that the chrysin according to the present invention or a pharmaceutically acceptable salt thereof promotes keratinocyte differentiation and increases the ability to retain moisture in the skin. Furthermore, the present invention is believed to be effective in preventing or improving skin dryness symptoms and atopic symptoms caused by incomplete epidermal differentiation of the skin, enhancing the flexibility and durability of the skin, Which is expected to exert an excellent effect on moisturizing the skin.
<실시예 3> 피부 장벽 기능 회복 강화 효과≪ Example 3 > Reinforcing effect of skin barrier function recovery
본 발명에 따른 조성물의 피부 장벽 기능 회복 효과를 알아보기 위하여 임상 시험을 실시하였다. 피험자들은 피험자 선정 및 제외기준에 따라 24세에서 48세(평균나이 30.6세)의 건강한 성인 남녀 27명을 선정하였으며 시험과 관련된 사항들을 사전에 구두로 공지하였고 자발적으로 참여의사를 밝힌 피험자를 대상으로 인체적용시험 참여 동의서를 받고 시험을 실시하였다. 모든 인체적용시험은 항온(22±2℃)항습(40~60%)조건에서 실시하였으며 측정 전 30분간 피험자들을 충분히 적응시킨 후 시험하였다. 수분손실량(TEWL: Transepidermal Water Loss)은 Tewameter TM210 (C & K. 독일)을 이용하여 측정하였으며 시험부위별로 5회 측정하여 평균값을 사용하였다.Clinical studies were conducted to examine the effect of the composition according to the present invention on restoring skin barrier function. Subjects were selected from 27 healthy adult men and women aged 24 to 48 years (mean age 30.6 years) according to the criteria for selection and exclusion of subjects. Subjects were informed in advance of the test related matters and volunteered to participate The test was conducted under the agreement of participation in the human body test. All human body tests were conducted at constant temperature (22 ± 2 ℃) and humidity (40 ~ 60%). Transepidermal water loss (TEWL) was measured using a Tewameter TM210 (C & K. Germany), and the mean value was used five times for each test site.
시험은 양쪽 전박(Ventral forearm)부위를 사용하였고 시험 시작 전 TEWL을 측정하여 장벽 파괴 전 base값으로 사용하였다. 증류수를 이용하여 1% SLS 용액을 조제하여 IQ chamber에 분주한 후 시험부위에 붙이고 24시간 동안 피부 장벽을 파괴하였다. 24시간 후 chamber를 제거하고 검정색으로 시험부위를 마킹하였으며 다음 날 TEWL을 측정하여 시료 도포 전 장벽 파괴 값으로 사용하였다. 시험 시료는 DPG에 3, 5, 10, 20 ㎍/ml 농도로 크리신을 녹여 사용하였다. 시료 무 처치 부위를 음성 대조군으로 사용하였다. 시료는 아침, 저녁 2회 시험 부위에 도포하였으며 시료 사용 1일, 3일, 7일, 14일 후 각각 TEWL을 측정하여 장벽 회복율을 계산하였다. 시험은 이중맹검법으로 수행하였다. 통계처리는 paired t-test를 통해 시료 사용 부위와 대조부위 간의 유의성 여부를 가설 평균 차 5%(p<0.05)로 확인하였으며 통계분석 프로그램은 마이크로소프트 엑셀 version 2010 소프트웨어를 이용하였다. 유효성 측면에서 시료 사용 부위와 대조 부위 간의 통계적으로 유의성이 있고 안전성 측면에서 사용기간 동안 제품에 의한 이상 반응이 발생하지 않은 경우 장벽회복 개선에 도움을 주는 시료로 판단하였다. 장벽 회복율은 아래 식에 따라 계산하였다.The ventral forearm was used for the test and the TEWL was measured before the start of the test and used as the base value before the barrier fracture. 1% SLS solution was prepared by using distilled water, placed in the IQ chamber, and placed on the test site to destroy the skin barrier for 24 hours. After 24 hours, the chamber was removed and the test area was marked with black. The next day, TEWL was measured and used as the barrier disruption value before sample application. The test samples were prepared by dissolving chrysin in DPG at concentrations of 3, 5, 10, and 20 ㎍ / ml. The sample-untreated site was used as a negative control. Samples were applied to the test site twice in the morning and evening, and TEWL was measured after 1 day, 3 days, 7 days, and 14 days, respectively. The test was performed by the double blind method. Statistical analysis was performed using a paired t-test. The significance level between the sample site and the control site was found to be 5% (p <0.05). Statistical analysis program was performed using Microsoft Excel version 2010 software. In terms of efficacy, there was a statistically significant difference between the site of use and the control site. From the safety standpoint, it was judged to be a sample to help improve the barrier recovery when no adverse reaction was caused by the product during the use period. The barrier recovery rate was calculated according to the following equation.
장벽 회복율(%) = [1-(시료 사용 x일 후 TEWL 측정 값-장벽 파괴 전 TEWL 측정 값)/(시료 도포 전 장벽 파괴 TEWL 측정 값-장벽 파괴 전 TEWL 측정 값]x100Barrier recovery rate (%) = [1- (TEWL measurement value after x days of sample use - TEWL measurement value before barrier destruction) / (TEWL measurement value before sample application - TEWL measurement value before barrier destruction) x100
시험시료에 대한 인체 적용 시험 결과는 상기 표 2에 나타낸 것처럼 10 ㎍/ml 크리신의 경우 도포 14일 후 85.4%(p<0.05), 20 ㎍/ml 크리신의 경우 도포 7일 후 75.1%(p<0.05), 도포 14일 후 85.0%(p<0.01)의 장벽 회복율을 나타내었으며 시료 무 처치 대조부위와 비교 시 통계적으로 유의한 수준을 나타내었다. 그리고 시료 사용기간 동안 어떠한 피부 유해사례가 관찰되지 않았다. 따라서 10 ㎍/ml 크리신은 도포 14일 후부터 20 ㎍/ml 크리신은 도포 7일 후부터 피부 장벽 회복 효과가 있는 것을 알 수 있었다. 따라서, 본 발명의 크리신 또는 이의 약학적으로 허용 가능한 염을 사용할 경우, 손상된 피부 장벽을 효과적으로 회복시킬 수 있을 것을 생각된다. 더 나아가 상기 결과로부터 본 발명의 조성물이 손상된 피부 장벽을 회복시켜 피부 본래의 보호기능을 원활히 수행함으로써 피부 건조성 질환에 효과적이며, 피부 건조로 인한 피부 잔주름 감소에도 효과가 있을 것으로 생각된다.As shown in Table 2, the results of human body application test were 85.4% (p <0.05) after 14 days for 10 ㎍ / ml chrysin and 75.1% (p <0.05) after 20 days for 20 ㎍ / 0.05). After 14 days of application, the barrier recovery rate was 85.0% (p <0.01), which was statistically significant compared with the untreated control site. No skin adverse events were observed during the sample period. Therefore, 10 ㎍ / ml chrysin was recovered from 14 days after application, and 20 ㎍ / ml chrysin was recovered from 7 days after application. Thus, it is contemplated that the use of the chrysin of the present invention or a pharmaceutically acceptable salt thereof will effectively restore the damaged skin barrier. Furthermore, from the above results, it can be said that the composition of the present invention restores the damaged skin barrier and smoothly carries out the original protective function of the skin, thereby being effective for skin dryness diseases and also reducing the fine lines caused by skin dryness.
<실시예 4> 피부 각질세포에서 자외선 조사로 인한 염증 반응 감소 효과<Example 4> Reduction of inflammatory reaction due to ultraviolet irradiation in dermal keratinocytes
본 발명에 따른 조성물의 피부 염증 완화 효과를 확인하기 위하여, 세포 수준에서 자외선에 대한 피부 각질 세포의 염증반응 감소 효과를 실험하였다. 인체 유래의 각질 세포에 25mJ/cm2 의 UVB를 조사하고, 크리신의 최종 농도가 1 ㎍/ml, 2㎍/ml이 되도록 각각 각질세포의 배양배지에 첨가하여 24 시간 배양한 후, 배양액을 수거하여 염증성 사이토카인으로 알려진 TNF-α의 양을 ELISA로 측정하였다. 이때, 효과의 비교를 위하여 아무것도 첨가하지 않은 각질세포의 배양배지(음성 대조군)에서도 동일한 방법으로 TNF-α 분비량을 측정하였다. TNF-α 분비량 증가율은 음성 대조군에 대한 분비량을 100%로 기준하여, 상대적인 분비량으로 계산하고 결과를 하기 표 3에 정리하였다.In order to confirm the skin irritation mitigating effect of the composition according to the present invention, the effect of decreasing the inflammatory response of keratinocytes against ultraviolet rays at the cell level was examined. The keratinocytes derived from the human body were irradiated with UVB at 25 mJ / cm 2 and added to the culture medium of keratinocytes so that the final concentration of chrysin was 1 / / ml and 2 / / ml, respectively. After culturing for 24 hours, The amount of TNF-a, known as an inflammatory cytokine, was measured by ELISA. At this time, in order to compare the effects, the secretion amount of TNF-α was also measured in the culture medium of the keratinocytes without any addition (negative control group) by the same method. The TNF-α secretion increase rate was calculated as the relative secretion amount based on the secretion amount of the negative control group as 100%, and the results are summarized in Table 3 below.
상기 표 3에 나타낸 바와 같이, 본 발명의 크리신은 음성 대조군보다 각질세포에서 자외선에 의한 자외선에 의한 TNF-α 분비량은 더 낮아지는 것을 확인할 수 있었다. 이러한 결과를 통해 본 발명의 조성물을 사용할 경우 피부 염증을 효과적으로 완화할 수 있을 것으로 기대된다. As shown in Table 3, it was confirmed that the amount of TNF-a secreted by ultraviolet rays from keratinocytes in keratin cells was lower than that in negative control cells of the present invention. These results suggest that the use of the composition of the present invention can effectively alleviate skin inflammation.
상기 실시예 1 내지 4의 결과에 비추어 볼 때, 크리신이 in vitro 상으로 비타민 D와 유사한 효과가 있다고 하더라도 실제 in vivo 상으로 피부 보습 효과, 피부 장벽 강화 효과, 피부 각질 세포 분화 효과 및/또는 피부 염증 완화 효과에 대해서는 본 발명의 실험을 통하여 확인할 수 있었다.In view of the results of Examples 1 to 4, even though the effect of chrysin is similar to that of vitamin D in vitro, it is possible to provide skin moisturizing effect, skin barrier strengthening effect, skin keratinocyte differentiation effect and / The anti-inflammatory effect was confirmed by the experiment of the present invention.
<제조예 1> 영양화장수 제조≪ Preparation Example 1 >
상기 표 4와 같이 크리신을 유효성분으로 포함하는 영양화장수를 통상의 방법에 따라 제조하였다. As shown in Table 4, a nutritional lotion containing crecine as an active ingredient was prepared according to a conventional method.
<제조예 2> 에센스 제조≪ Preparation Example 2 > Preparation of essence
상기 표 5와 같이 크리신을 유효성분으로 포함하는 에센스를 통상의 방법에 따라 제조하였다. Essences containing chrysin as an active ingredient were prepared according to a conventional method as shown in Table 5 above.
<제조예 3> 크림 제조≪ Preparation Example 3 >
상기 표 6와 같이 크리신을 유효성분으로 포함하는 크림을 통상의 방법에 따라 제조하였다. As shown in Table 6, a cream containing chrysin as an active ingredient was prepared by a conventional method.
<제조예 4> 팩의 제조≪ Preparation Example 4 > Preparation of pack
상기 표 7와 같이 크리신을 유효성분으로 포함하는 팩을 통상의 방법에 따라 제조하였다. As shown in Table 7, a pack containing chrysin as an active ingredient was prepared according to a conventional method.
<제조예 5> 피부 외용제 중 연고의 제조Preparation Example 5 Preparation of ointment for external preparation for skin
상기 표 8와 같이 크리신을 유효성분으로 포함하는 피부 외용제 중 연고를 통상의 방법에 따라 제조하였다.As shown in Table 8, ointments among external preparations for skin containing chrysin as an active ingredient were prepared according to a conventional method.
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