KR102631500B1 - Composition for skin whitening comprising extract of hempseed meal as effective component - Google Patents
Composition for skin whitening comprising extract of hempseed meal as effective component Download PDFInfo
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- KR102631500B1 KR102631500B1 KR1020210025782A KR20210025782A KR102631500B1 KR 102631500 B1 KR102631500 B1 KR 102631500B1 KR 1020210025782 A KR1020210025782 A KR 1020210025782A KR 20210025782 A KR20210025782 A KR 20210025782A KR 102631500 B1 KR102631500 B1 KR 102631500B1
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- Prior art keywords
- seed meal
- extract
- hemp seed
- hemp
- composition
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- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
- A61K8/9783—Angiosperms [Magnoliophyta]
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- 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
- A23L33/105—Plant extracts, their artificial duplicates or their derivatives
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- A—HUMAN NECESSITIES
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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
본 발명은 헴프씨드박 추출물을 유효성분으로 함유하는 피부 미백용 조성물에 관한 것으로, 상기 유효성분은 티로시나아제 저해 활성과 멜라닌 합성 억제 활성이 우수하므로, 피부 미백용 조성물 및 멜라닌 색소 과다 침착 질환의 예방, 개선 또는 치료용 조성물로 유용하게 사용될 수 있다. The present invention relates to a composition for skin whitening containing hemp seed meal extract as an active ingredient. The active ingredient has excellent tyrosinase inhibitory activity and melanin synthesis inhibitory activity, and is therefore effective in treating skin whitening composition and melanin hyperpigmentation disease. It can be usefully used as a composition for prevention, improvement or treatment.
Description
본 발명은 헴프씨드박 추출물을 유효성분으로 함유하는 피부 미백용 조성물에 관한 것이다.The present invention relates to a composition for skin whitening containing hemp seed meal extract as an active ingredient.
표피 기저층의 멜라닌 세포는 피부의 다양한 자극 요인에 의해 멜라닌을 만들고, 이 멜라닌이 각질층에 도달하여 과색소 침착증을 유발하면 피부가 검게 보인다. 멜라닌 합성에 관여하는 요인으로는 난포 호르몬, 황체 호르몬, 색소 세포 자극 호르몬, 자외선, 스트레스 그리고 유전 등이 있다. Melanocytes in the basal layer of the epidermis produce melanin in response to various irritating factors in the skin, and when this melanin reaches the stratum corneum and causes hyperpigmentation, the skin appears dark. Factors involved in melanin synthesis include follicular hormone, progesterone, pigment cell-stimulating hormone, ultraviolet rays, stress, and heredity.
불필요해진 멜라닌 색소를 재빨리 배출해서 그 사람이 지니고 있는 본래 피부의 투명함을 되찾는 것을 피부 미백이라 한다. 식품의약품안전청에 미백 성분으로 등록된 물질은 닥나무 추출물, 알부틴, 에틸아스코빌에텔, 유용성 감초 추출물, 아스코빌글루코사이드, 나이아신아마이드, 알파-비사볼올 및 아스코빌테트라이소팔미테이트 등이 있으며, 이와 같은 물질을 이용해 로션이나 액제, 크림, 마스크팩 등으로 만드는 것이 미백 화장품이다.Skin whitening refers to the process of quickly discharging unnecessary melanin pigment and restoring the original transparency of the person's skin. Substances registered as whitening ingredients with the Food and Drug Administration include mulberry extract, arbutin, ethyl ascorbyl ether, oil-soluble licorice extract, ascorbyl glucoside, niacinamide, alpha-bisabolol, and ascorbyl tetraisopalmitate. Whitening cosmetics are made into lotions, liquids, creams, mask packs, etc.
닥나무 추출물, 알부틴, 알파-비사볼올 및 유용성 감초 추출물은 멜라닌 합성에 관여하는 티로시나아제 효소가 활성화되는 것을 억제하고, 아스코빌글루코사이드, 에틸아스코빌에텔 및 아스코빌테트라이소팔미테이트는 티로시나아제 효소에 자극받은 티로신이 산화되는 것을 억제하며, 나이아신아마이드는 이미 생성된 멜라닌이 멜라닌 세포에서 각질형성세포로 넘어가는 단계를 억제하여 멜라닌이 실제 피부 세포에 들어가는 마지막 단계를 막는다.Mulberry extract, arbutin, alpha-bisabolol, and oil-soluble licorice extract inhibit the activation of the tyrosinase enzyme involved in melanin synthesis, and ascorbyl glucoside, ethyl ascorbyl ether, and ascorbyl tetraisopalmitate inhibit the tyrosinase enzyme. It inhibits the oxidation of stimulated tyrosine, and niacinamide inhibits the transition of already produced melanin from melanocytes to keratinocytes, preventing the final step of melanin entering actual skin cells.
미용이 아닌 의료 목적으로 사용되는 대표적인 물질은 의약품으로 처방되는 하이드로퀴논이며, 주로 피부 연고로 사용된다. 한국에서 이 물질이 4% 이상 함유된 연고를 구입하려면 반드시 의사의 처방이 있어야 하며, 2% 연고는 처방전 없이도 약국에서 구입할 수 있다. 의료 목적으로 사용되는 하이드로퀴논 연고는 미백 화장품과는 다르게 멜라닌이 있는 피부 세포를 직접 파괴해 새로운 피부 세포가 자라도록 돕는다.A representative substance used for medical purposes other than cosmetics is hydroquinone, which is prescribed as a medicine and is mainly used as a skin ointment. In Korea, to purchase an ointment containing more than 4% of this substance, you must have a doctor's prescription, and 2% ointment can be purchased at a pharmacy without a prescription. Unlike whitening cosmetics, hydroquinone ointment used for medical purposes directly destroys skin cells containing melanin and helps new skin cells grow.
하지만, 상기 물질들은 비교적 높은 농도에서만 티로시나아제 활성 저해 효과가 나타나거나, 안정성이 낮아 효과가 오래 지속되지 못하는 등 제품의 적용 범위가 좁다는 문제점이 있으며, 하이드로퀴논은 피부자극 및 안정성 문제로 인하여 화장료에서의 사용이 제한되고 있다.However, the above substances have the problem of narrow product application scope, such as the effect of inhibiting tyrosinase activity only at relatively high concentrations or the effect does not last long due to low stability, and hydroquinone has skin irritation and stability issues. Its use in cosmetics is limited.
한편, 헴프씨드박은 헴프씨드(hempseed)로부터 기름을 짜고 남은 가공부산물로, 탈지 헴프씨드(defatted hempseed)라고도 한다. 헴프씨드는 삼과의 한해살이풀인 대마의 씨앗으로, 대마의 잎과 꽃에서 얻어지는 대마초에 비해 마취 성분인 테트라히드로카나비놀(tetrahydrocannabinol, THC) 함량이 2% 미만으로 낮은 편이다. 한방에서는 마인(麻仁), 마자인(麻子仁)이라 하여 껍질이 제거된 상태에서 약으로 처방된다. 헴프씨드에 함유된 오일은 불포화지방산으로, 양질의 단백질이 다량 함유되어 있고, 동맥경화와 같은 심혈관계 질환 예방에 도움을 주며 섬유질이 풍부하여 소화에 좋고 변비 예방에도 효과적이다. 헴프씨드에 대한 관심이 높아지면서 헴프씨드로부터 유지 성분을 추출하고 남은 부산물을 이용한 연구가 이루어져 왔으며, 이러한 탈지 헴프씨드에는 페놀산(phenolic acid), 리그난(lignan), 플라보노이드(flavonoid) 등이 많이 함유되어 있다고 보고된 바 있다. Meanwhile, hemp seed meal is a processing by-product left after extracting oil from hempseed, and is also called defatted hempseed. Hemp seeds are the seeds of hemp, an annual herb from the hemp family. Compared to cannabis obtained from the leaves and flowers of hemp, the content of tetrahydrocannabinol (THC), an anesthetic ingredient, is lower at less than 2%. In oriental medicine, it is called ma-in (麻仁) or maja-in (麻子仁) and is prescribed as a medicine with the peel removed. The oil contained in hemp seeds is an unsaturated fatty acid, contains a large amount of high-quality protein, helps prevent cardiovascular diseases such as arteriosclerosis, and is rich in fiber, which is good for digestion and effective in preventing constipation. As interest in hemp seeds has increased, research has been conducted using the by-products remaining after extracting the fat content from hemp seeds. These defatted hemp seeds contain a lot of phenolic acid, lignans, and flavonoids. It has been reported that it has been done.
한편, 한국공개특허 제2013-0113581호에는 '헴프씨 추출물을 포함하는 동맥경화증 예방 및 치료용 조성물'이 개시되어 있고, 한국등록특허 제1997166호에는 '헴프씨드 추출물을 포함하는 항여드름, 여드름 흉터개선, 주름 및 탄력 개선, 피부재생, 피지억제용 조성물'이 개시되어 있으나, 본 발명의 '헴프씨드박 추출물을 유효성분으로 함유하는 피부 미백용 조성물'에 대해서는 기재된 바가 없다.Meanwhile, Korean Patent Publication No. 2013-0113581 discloses 'Composition for preventing and treating arteriosclerosis containing hemp seed extract', and Korean Patent No. 1997166 discloses 'Anti-acne and acne scarring containing hemp seed extract'. A composition for improving wrinkles and elasticity, improving skin regeneration, and suppressing sebum' is disclosed, but there is no description of the 'composition for skin whitening containing hemp seed meal extract as an active ingredient' of the present invention.
본 발명은 상기와 같은 요구에 의해 도출된 것으로서, 본 발명은 헴프씨드박 추출물을 유효성분으로 함유하는 피부 미백용 조성물을 제공하고, 상기 유효성분이 멜라노마 세포에서 티로시나아제 저해 활성 및 멜라닌 합성 억제 활성이 있다는 것을 확인함으로써, 본 발명을 완성하였다.The present invention was developed in response to the above-mentioned needs. The present invention provides a composition for skin whitening containing hemp seed meal extract as an active ingredient, and the active ingredient has tyrosinase inhibitory activity and inhibition of melanin synthesis in melanoma cells. By confirming that it is active, the present invention was completed.
상기 과제를 해결하기 위해, 본 발명은 헴프씨드박 추출물을 유효성분으로 함유하는 피부 미백용 화장료 조성물을 제공한다. In order to solve the above problems, the present invention provides a cosmetic composition for skin whitening containing hemp seed meal extract as an active ingredient.
또한, 본 발명은 헴프씨드박 추출물을 유효성분으로 함유하는 멜라닌 색소 과다 침착 질환의 예방 또는 개선용 건강기능식품 조성물을 제공한다. In addition, the present invention provides a health functional food composition for preventing or improving melanin hyperpigmentation disease containing hemp seed meal extract as an active ingredient.
또한, 본 발명은 헴프씨드박 추출물을 유효성분으로 함유하는 멜라닌 색소 과다 침착 질환의 예방 또는 치료용 약학 조성물을 제공한다. In addition, the present invention provides a pharmaceutical composition for preventing or treating melanin hyperpigmentation disease containing hemp seed meal extract as an active ingredient.
본 발명은 헴프씨드박 추출물을 유효성분으로 함유하는 피부 미백용 조성물에 관한 것으로, 상기 유효성분은 티로시나아제 저해 활성과 멜라닌 합성 억제 활성이 우수하므로, 피부 미백용 조성물 및 멜라닌 색소 과다 침착 질환의 예방 또는 치료용 조성물로 유용하게 사용될 수 있다. The present invention relates to a composition for skin whitening containing hemp seed meal extract as an active ingredient. The active ingredient has excellent tyrosinase inhibitory activity and melanin synthesis inhibitory activity, and is therefore effective in treating skin whitening composition and melanin hyperpigmentation disease. It can be usefully used as a preventive or therapeutic composition.
도 1은 헴프씨드박 추출물의 처리에 따른 멜라노마 세포(B16F10)에서의 세포 독성을 확인한 결과이다. CON은 아무것도 처리하지 않은 대조군이다.
도 2는 헴프씨드박 추출물의 처리에 따른 멜라노마 세포(B16F10)에서의 멜라닌 합성량을 나타낸 결과이다. CON은 아무것도 처리하지 않은 정상군이고, arbutin은 양성대조군이며,α-MSH는 멜라닌 합성 유도제이다. 도면 내 서로 다른 문자 a~f는 서로 유의미한 차이가 있다는 것을 의미하며, p<0.05이다.
도 3은 헴프씨드박 추출물의 처리에 따른 멜라노마 세포(B16F10)에서의 티로시나아제 활성을 나타낸 결과이다. CON은 아무것도 처리하지 않은 정상군이고, arbutin은 양성대조군이며,α-MSH는 티로시나아제 활성 유도제이다. 도면 내 서로 다른 문자 a~f는 서로 유의미한 차이가 있다는 것을 의미하며, p<0.05이다.
도 4는 헴프씨드박 추출물의 처리에 따른 멜라노마 세포(B16F10)에서의 MITF(microphthalmia-associated transcription factor) 유전자 발현양을 나타낸 결과이다. CON은 아무것도 처리하지 않은 정상군이고, arbutin은 양성대조군이며,α-MSH는 멜라닌 합성 유도제이다. 도면 내 서로 다른 문자 a~f는 서로 유의미한 차이가 있다는 것을 의미하며, p<0.05이다.Figure 1 shows the results of confirming cytotoxicity in melanoma cells (B16F10) according to treatment with hemp seed meal extract. CON is the control group with no treatment.
Figure 2 is a result showing the amount of melanin synthesis in melanoma cells (B16F10) according to treatment with hemp seed meal extract. CON is the normal group without any treatment, arbutin is the positive control group, and α-MSH is a melanin synthesis inducer. Different letters a to f in the figure mean that there is a significant difference from each other, p<0.05.
Figure 3 shows the results showing tyrosinase activity in melanoma cells (B16F10) according to treatment with hemp seed meal extract. CON is a normal group without any treatment, arbutin is a positive control group, and α-MSH is a tyrosinase activity inducer. Different letters a to f in the figure mean that there is a significant difference from each other, p<0.05.
Figure 4 shows the results showing the level of MITF (microphthalmia-associated transcription factor) gene expression in melanoma cells (B16F10) according to treatment with hemp seed meal extract. CON is the normal group without any treatment, arbutin is the positive control group, and α-MSH is a melanin synthesis inducer. Different letters a to f in the figure mean that there is a significant difference from each other, p<0.05.
본 발명의 목적을 달성하기 위하여, 본 발명은 헴프씨드박 추출물을 유효성분으로 함유하는 피부 미백용 화장료 조성물을 제공한다. In order to achieve the object of the present invention, the present invention provides a cosmetic composition for skin whitening containing hemp seed meal extract as an active ingredient.
상기 헴프씨드박 추출물의 용매는 물, C1~C4의 저급 알코올 또는 이들의 혼합물일 수 있으며, 바람직하게는 에탄올을 용매로 사용하여 추출한 것이나, 이에 제한되지 않는다.The solvent for the hemp seed meal extract may be water, a lower alcohol of C 1 to C 4 , or a mixture thereof, and is preferably extracted using ethanol as a solvent, but is not limited thereto.
상기 헴프씨드박은 헴프씨드로부터 기름을 착유하고 남은 부산물이다.The hemp seed meal is a by-product remaining after extracting oil from hemp seeds.
본 발명의 일 구현 예에서, 상기 헴프씨드박 추출물은 티로시나아제의 활성을 억제하거나, 멜라닌 합성을 억제하는 효과가 있다. In one embodiment of the present invention, the hemp seed meal extract has the effect of inhibiting the activity of tyrosinase or inhibiting melanin synthesis.
본 발명의 일 구현예에 따른 화장료 조성물에 있어서, 상기 피부 미백용 화장료 조성물은 크림, 유연화장수, 영양화장수, 팩, 에센스, 헤어토닉, 샴푸, 린스, 헤어 컨디셔너, 헤어 트리트먼트, 젤, 스킨로션, 스킨소프너, 스킨토너, 아스트린젠트, 밀크로션, 모이스처 로션, 영양로션, 마사지 크림, 영양크림, 모이스처 크림, 핸드 크림, 파운데이션, 영양에센스, 선스크린, 비누, 클렌징폼, 클렌징로션, 클렌징크림, 바디 로션 및 바디 클렌저로 이루어지는 군으로부터 선택된 어느 하나의 제형을 가질 수 있으나, 이에 제한되지 않는다. 이들 각 제형으로 이루어진 화장료 조성물은 그 제형의 제제화에 필요하고 적절한 각종의 기제와 첨가물을 함유할 수 있으며, 이들 성분의 종류와 양은 당업자에 의해 용이하게 선정될 수 있다. In the cosmetic composition according to one embodiment of the present invention, the cosmetic composition for skin whitening is cream, softening lotion, nourishing lotion, pack, essence, hair tonic, shampoo, rinse, hair conditioner, hair treatment, gel, skin lotion. , skin softener, skin toner, astringent, milk lotion, moisture lotion, nutritional lotion, massage cream, nutritional cream, moisture cream, hand cream, foundation, nutritional essence, sunscreen, soap, cleansing foam, cleansing lotion, cleansing cream, body. It may have any one formulation selected from the group consisting of lotion and body cleanser, but is not limited thereto. A cosmetic composition composed of each of these formulations may contain various bases and additives necessary and appropriate for the formulation of the formulation, and the types and amounts of these components can be easily selected by a person skilled in the art.
본 발명의 화장료 조성물의 제형이 페이스트, 크림 또는 겔인 경우에는 담체 성분으로서 동물섬유, 식물섬유, 왁스, 파라핀, 전분, 트라가칸트, 셀룰로오스 유도체, 폴리에틸렌 글리콜, 실리콘, 벤토나이트, 실리카, 탈크 또는 산화아연 등이 이용될 수 있다.When the formulation of the cosmetic composition of the present invention is a paste, cream, or gel, animal fiber, plant fiber, wax, paraffin, starch, tragacanth, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc, or zinc oxide are used as carrier ingredients. etc. can be used.
본 발명의 화장료 조성물의 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 cosmetic composition of the present invention is powder or spray, lactose, talc, silica, aluminum hydroxide, calcium silicate, or polyamide powder can be used as the carrier ingredient. In particular, when the cosmetic composition is a spray, chlorofluorohydride may be used as a carrier ingredient. It may contain propellants such as carbon, propane-butane or dimethyl ether.
본 발명의 화장료 조성물의 제형이 용액 또는 유탁액의 경우에는 담체 성분으로서 용매, 용매화제 또는 유탁화제가 이용되고, 예컨대 물, 에탄올, 이소프로판올, 에틸 카보네이트, 에틸 아세테이트, 벤질 알코올, 벤질 벤조에이트, 프로필렌글리콜, 1,3-부틸글리콜 오일, 글리세롤 지방족 에스테르, 폴리에틸렌 글리콜 또는 소르비탄의 지방산 에스테르가 있다.When the formulation of the cosmetic composition 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. These include fatty acid esters of glycol, 1,3-butylglycol oil, glycerol aliphatic esters, polyethylene glycol or sorbitan.
본 발명의 화장료 조성물의 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 cosmetic composition of the present invention is a suspension, the carrier component includes water, a liquid diluent such as ethanol or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester, and polyoxyethylene sorbitan ester, Microcrystalline cellulose, aluminum metahydroxide, bentonite, agar, or tracant may be used.
본 발명의 화장료 조성물의 제형이 계면-활성제 함유 클렌징인 경우에는 담체 성분으로서 지방족 알코올 설페이트, 지방족 알코올 에테르설페이트, 설포숙신산 모노에스테르, 아세티오네이트, 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 cosmetic composition of the present invention is a surfactant-containing cleansing agent, the carrier ingredients include aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, acethionate, imidazolinium derivative, methyl taurate, and sarcosinate. , fatty acid amide ether sulfate, alkylamidobetaine, fatty alcohol, fatty acid glyceride, fatty acid diethanolamide, vegetable oil, lanolin derivative, or ethoxylated glycerol fatty acid ester can be used.
본 발명의 화장료 조성물은 형광물질, 살진균제, 굴수성 유발물질, 보습제, 방향제, 방향제 담체, 단백질, 용해화제, 당 유도체, 일광차단제, 비타민, 식물 추출물 등을 포함하는 부형제를 추가로 함유할 수 있다.The cosmetic composition of the present invention may further contain excipients including fluorescent substances, fungicides, hydrotropes-inducing substances, moisturizers, fragrances, fragrance carriers, proteins, solubilizers, sugar derivatives, sunscreens, vitamins, plant extracts, etc. .
또한, 본 발명은 헴프씨드박 추출물을 유효성분으로 함유하는 멜라닌 색소 과다 침착 질환의 예방 또는 개선용 건강기능식품 조성물을 제공한다. In addition, the present invention provides a health functional food composition for preventing or improving melanin hyperpigmentation disease containing hemp seed meal extract as an active ingredient.
본 발명의 건강기능식품 조성물은 이너뷰티 푸드(inner beauty food)의 형태로 섭취함으로써 더욱 우수한 효과를 갖는 장점을 가진다. 상기 이너뷰티는 '먹는 화장품 또는 뷰티 푸드'로 일컬어지는 푸드로, 피부에 좋은 여러 가지 성분을 몸속으로 흡수시켜 피부 체질을 건강하게 바꾸는 식품을 지칭하며, 피부 타입에 맞는 화장품을 고르듯 피부 컨디션과 라이프스타일을 고려해 개개인에게 맞는 이너뷰티 푸드를 선택하여 섭취할 수 있다. 전술한 화장료 조성물을 포함하는 화장품과 본 발명의 화합물을 포함하는 이너뷰티 푸드를 혼용할 경우, 화장품만 사용하는 것에 비해 피부 미백 효과가 월등히 높아져 멜라닌 색소 과다 침착 질환에 더욱 효과적일 수 있다.The health functional food composition of the present invention has the advantage of having more excellent effects when consumed in the form of inner beauty food. The inner beauty is a food called 'edible cosmetics or beauty food'. It refers to foods that change the skin constitution to a healthy one by absorbing various ingredients that are good for the skin into the body. Just like choosing cosmetics that suit your skin type, it is important to check your skin condition and health. Considering your lifestyle, you can choose and consume inner beauty foods that suit each individual. When cosmetics containing the above-described cosmetic composition are mixed with inner beauty food containing the compound of the present invention, the skin whitening effect is significantly increased compared to using only cosmetics, making it more effective for melanin hyperpigmentation disease.
본 발명의 건강기능식품 조성물은 분말, 과립, 환, 정제, 캡슐, 캔디, 시럽 및 음료 중에서 선택된 하나의 제형일 수 있으나, 이에 제한되지 않는다. The health functional food composition of the present invention may be in a formulation selected from powder, granule, pill, tablet, capsule, candy, syrup, and beverage, but is not limited thereto.
본 발명의 건강기능식품 조성물을 식품첨가물로 사용하는 경우, 상기 건강기능식품 조성물을 그대로 첨가하거나 다른 식품 또는 식품성분과 함께 사용될 수 있고, 통상적인 방법에 따라 적절하게 사용될 수 있다. 유효 성분의 혼합양은 그의 사용 목적(예방 또는 개선)에 따라 적절하게 사용될 수 있다. 일반적으로, 식품 또는 음료의 제조시 본 발명의 건강기능식품 조성물은 원료에 대하여 15 중량부 이하, 바람직하게는 10 중량부 이하의 양으로 첨가된다. 그러나 건강을 목적으로 하는 장기간의 섭취의 경우에는 상기 양은 상기 범위 이하일 수 있으며, 안전성 면에서 아무런 문제가 없기 때문에 유효성분은 상기 범위 이상의 양으로 사용될 수 있다.When using the health functional food composition of the present invention as a food additive, the health functional food composition may be added as is or used together with other foods or food ingredients, and may be used appropriately according to conventional methods. The mixed amount of active ingredients can be appropriately used depending on the purpose of use (prevention or improvement). Generally, when manufacturing a food or beverage, the health functional food composition of the present invention is added in an amount of 15 parts by weight or less, preferably 10 parts by weight or less, based on the raw materials. However, in the case of long-term intake for health purposes, the amount may be below the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount above the above range.
상기 건강기능식품의 종류에 특별한 제한은 없다. 상기 건강기능식품 조성물을 첨가할 수 있는 식품의 예로는 육류, 소시지, 빵, 초콜릿, 캔디류, 스낵류, 과자류, 피자, 라면, 기타 면류, 껌류, 아이스크림류를 포함한 낙농제품, 각종 스프, 음료수, 차 드링크제, 알콜 음료 및 비타민 복합제 등이 있으며, 통상적인 의미에서의 건강식품을 모두 포함한다.There are no particular restrictions on the types of health functional foods. Examples of foods to which the health functional food composition can be added include meat, sausages, bread, chocolate, candies, snacks, confectionery, pizza, ramen, other noodles, gum, dairy products including ice cream, various soups, beverages, and tea. There are drinks, alcoholic beverages, and vitamin complexes, and it includes all health foods in the conventional sense.
또한, 본 발명의 건강기능식품 조성물은 식품, 특히 기능성 식품으로 제조될 수 있다. 본 발명의 기능성 식품은 식품 제조 시에 통상적으로 첨가되는 성분을 포함하며, 예를 들어, 단백질, 탄수화물, 지방, 영양소 및 조미제를 포함한다. 예컨대, 드링크제로 제조되는 경우에는 유효성분 이외에 천연 탄수화물 또는 향미제를 추가 성분으로서 포함시킬 수 있다. 상기 천연 탄수화물은 모노사카라이드(예컨대, 글루코오스, 프럭토오스 등), 디사카라이드(예컨대, 말토스, 수크로오스 등), 올리고당, 폴리사카라이드(예컨대, 덱스트린, 시클로덱스트린 등), 또는 당알코올(예컨대, 자일리톨, 소르비톨, 에리쓰리톨 등)인 것이 바람직하다. 상기 향미제는 천연 향미제(예컨대, 타우마틴, 스테비아 추출물 등)와 합성 향미제(예컨대, 사카린, 아스파르탐 등)를 이용할 수 있다.Additionally, the health functional food composition of the present invention can be manufactured into food, especially functional food. The functional food of the present invention includes ingredients commonly added during food production, and includes, for example, proteins, carbohydrates, fats, nutrients and seasonings. For example, when manufacturing a drink, natural carbohydrates or flavoring agents may be included as additional ingredients in addition to the active ingredient. The natural carbohydrates include monosaccharides (e.g., glucose, fructose, etc.), disaccharides (e.g., maltose, sucrose, etc.), oligosaccharides, polysaccharides (e.g., dextrins, cyclodextrins, etc.), or sugar alcohols (e.g., For example, xylitol, sorbitol, erythritol, etc.) is preferable. The flavoring agent may be a natural flavoring agent (e.g., thaumatin, stevia extract, etc.) or a synthetic flavoring agent (e.g., saccharin, aspartame, etc.).
상기 건강기능식품 조성물 이외에 여러 가지 영양제, 비타민, 전해질, 풍미제, 착색제, 펙트산 및 그의 염, 알긴산 및 그의 염, 유기산, 보호성 콜로이드 증점제, pH 조절제, 안정화제, 방부제, 글리세린, 알콜, 탄산음료에 사용되는 탄산화제 등을 더 함유할 수 있다. 이러한 상기 첨가되는 성분의 비율은 크게 중요하진 않지만 본 발명의 건강기능품 조성물 100 중량부에 대하여, 0.01 내지 0.1 중량부의 범위에서 선택되는 것이 일반적이다.In addition to the above health functional food composition, various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, carbonic acid. It may further contain carbonating agents used in beverages. The ratio of the ingredients added is not very important, but is generally selected in the range of 0.01 to 0.1 parts by weight based on 100 parts by weight of the health functional product composition of the present invention.
또한, 본 발명은 헴프씨드박 추출물을 유효성분으로 함유하는 멜라닌 색소 과다 침착 질환의 예방 또는 치료용 약학 조성물을 제공한다. In addition, the present invention provides a pharmaceutical composition for preventing or treating melanin hyperpigmentation disease containing hemp seed meal extract as an active ingredient.
상기 멜라닌 색소 과다 침착(hyperpigmentation)은 피부 또는 손·발톱의 특정 부위에서 멜라닌의 과도한 증가에 의해 다른 부위에 비해 검게 또는 어둡게 되는 것을 의미한다. 상기 멜라닌 색소 과다 침착 질환은 주근깨, 노인성 반점, 간반, 기미, 갈색 또는 흑점, 일광 색소반, 푸른흑피증(cyanic melasma), 약물 사용 후의 과다색소침착, 임신성 갈색반(gravidic chloasma) 또는 찰상 및 화상을 비롯한 상처 또는 피부염으로 인한 염증 후 과다 색소 침착 등을 포함하나 이에 한정되지 않는다. The hyperpigmentation of melanin means that a specific area of the skin or hands or toenails becomes black or darker than other areas due to an excessive increase in melanin. The above melanin hyperpigmentation diseases include freckles, senile spots, liver spots, freckles, brown or black spots, solar pigment spots, cyanic melasma, hyperpigmentation after drug use, gravidic chloasma, or scratches and burns. This includes, but is not limited to, hyperpigmentation after inflammation due to wounds or dermatitis.
본 발명의 약학 조성물은 약학적 조성물의 제조에 통상적으로 사용하는 적절한 담체, 부형제 또는 희석제를 더 포함할 수 있다. 본 발명에 따른 조성물의 약학적 투여 형태는 단독으로 또는 타 약학적 활성 화합물과 결합뿐만 아니라 적당한 조합으로 사용될 수 있다.The pharmaceutical composition of the present invention may further include appropriate carriers, excipients, or diluents commonly used in the preparation of pharmaceutical compositions. The pharmaceutical dosage form of the composition according to the present invention can be used alone or in combination with other pharmaceutically active compounds, as well as in appropriate combinations.
본 발명에 따른 약학 조성물은, 각각 통상의 방법에 따라 산제, 과립제, 정제, 캡슐제, 현탁액, 에멀젼, 시럽, 에어로졸 등의 경구형 제제, 외용제, 좌제 및 주사제의 형태로 제형화하여 사용될 수 있다. 상기 헴프씨드박 추출물을 포함하는 약학 조성물에 포함될 수 있는 담체, 부형제 및 희석제로는 락토즈, 덱스트로즈, 수크로스, 솔비톨, 만니톨, 자일리톨, 에리스리톨, 말티톨, 전분, 아카시아 고무, 알지네이트, 젤라틴, 칼슘 포스페이트, 칼슘 실리케이트, 셀룰로오스, 메틸 셀룰로오스, 미정질 셀룰로오스, 폴리비닐 피롤리돈, 물, 메틸히드록시벤조에이트, 프로필히드록시벤조에이트, 탈크, 마그네슘 스테아레이트 및 광물유 등을 포함한 다양한 화합물 혹은 혼합물을 들 수 있다. 제제화할 경우에는 보통 사용하는 충진제, 증량제, 결합제, 습윤제, 붕해제, 계면활성제 등의 희석제 또는 부형제를 사용하여 조제된다. 경구 투여를 위한 고형 제제에는 정제, 환제, 산제, 과립제, 캡슐제 등이 포함되며, 이러한 고형 제제는 상기 헴프씨드박 추출물에 적어도 하나 이상의 부형제 예를 들면, 전분, 칼슘 카보네이트, 수크로오스 또는 락토오스, 젤라틴 등을 섞어 조제된다. 또한 단순한 부형제 이외에 마그네슘 스테아레이트, 탈크 같은 윤활제들도 사용된다. 경구를 위한 액상 제제로는 현탁제, 내용액제, 유제, 시럽제 등이 해당되는데 흔히 사용되는 단순 희석제인 물, 리퀴드 파라핀 이외에 여러 가지 부형제, 예를 들면 습윤제, 감미제, 방향제, 보존제 등이 포함될 수 있다. 비경구 투여를 위한 제제에는 멸균된 수용액, 비수성용제, 현탁제, 유제, 동결건조 제제, 좌제가 포함된다. 비수성용제, 현탁제로는 프로필렌글리콜, 폴리에틸렌 글리콜, 올리브 오일과 같은 식물성 기름, 에틸올레이트와 같은 주사 가능한 에스테르 등이 사용될 수 있다. 좌제의 기제로는 위텝솔, 마크로골, 트윈 61, 카카오지, 라우린지, 글리세로젤라틴 등이 사용될 수 있다.The pharmaceutical composition according to the present invention can be formulated and used in the form of oral preparations such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, external preparations, suppositories, and injections according to conventional methods. . Carriers, excipients, and diluents that may be included in the pharmaceutical composition containing the hemp seed meal extract include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, Various compounds or mixtures including calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. I can hear it. When formulated, it is prepared using diluents or excipients such as commonly used fillers, extenders, binders, wetting agents, disintegrants, and surfactants. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc. These solid preparations include the hemp seed meal extract with at least one excipient, such as starch, calcium carbonate, sucrose or lactose, and gelatin. It is prepared by mixing etc. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Liquid preparations for oral use include suspensions, oral solutions, emulsions, syrups, etc. In addition to the commonly used simple diluents such as water and liquid paraffin, various excipients such as wetting agents, sweeteners, fragrances, and preservatives may be included. . Preparations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, lyophilized preparations, and suppositories. Non-aqueous solvents and suspensions include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, and injectable ester such as ethyl oleate. As a base for suppositories, wethepsol, macrogol, Tween 61, cacao, laurin, glycerogelatin, etc. can be used.
본 발명의 약학 조성물의 바람직한 투여량은 제제화 방법, 투여 방식, 환자의 연령, 체중, 성, 병적 상태, 음식, 투여 시간, 투여 경로, 배설 속도 및 반응 감응성과 같은 요인들에 의해 다양하게 처방될 수 있다. The preferred dosage of the pharmaceutical composition of the present invention may be prescribed in various ways depending on factors such as formulation method, administration method, patient's age, weight, sex, pathological condition, food, administration time, administration route, excretion rate, and reaction sensitivity. You can.
본 발명의 약학 조성물은 경구 또는 비경구로 투여할 수 있으며, 비경구 투여의 경우, 피부에 국소적으로 도포, 정맥 내 주입, 피하 주입, 근육 주입, 복강 주입, 경피 투여 등으로 투여할 수 있다. The pharmaceutical composition of the present invention can be administered orally or parenterally, and in the case of parenteral administration, it can be administered topically on the skin, intravenous injection, subcutaneous injection, intramuscular injection, intraperitoneal injection, transdermal administration, etc.
이하, 본 발명을 실시예에 의해 더욱 상세히 설명한다. 이들 실시예는 오로지 본 발명을 보다 구체적으로 설명하기 위한 것으로 본 발명의 범위가 이들에 의해 제한되지 않는다는 것은 당해 기술분야에서 통상의 지식을 가진 자에게 있어 자명한 것이다.Hereinafter, the present invention will be explained in more detail by examples. These examples are only for illustrating the present invention in more detail, and it is obvious to those skilled in the art that the scope of the present invention is not limited thereto.
제조예 1. 헴프씨드박 추출물의 제조Preparation Example 1. Preparation of hemp seed meal extract
헴프씨드는 (주)어썸티에서 구매하여 사용하였다. 헴프씨드 10g에 100㎖의 헥산(hexane)을 첨가하여 10,000 rpm으로 10분 동안 균질화하고, 실온에서 180 rpm으로 3시간 동안 교반하여 추출한 후, 12,000 rpm으로 10분 동안 원심분리하여 상등액(오일 성분)을 제거하였다. 이후, 남은 잔사를 30℃에서 24시간 동안 건조시켜, 총 4.7g의 헴프씨드박을 회수하였다. Hemp seeds were purchased and used from Awesome Tea Co., Ltd. Add 100 ml of hexane to 10 g of hemp seeds, homogenize at 10,000 rpm for 10 minutes, extract by stirring at room temperature for 3 hours at 180 rpm, and then centrifuge at 12,000 rpm for 10 minutes to obtain supernatant (oil component). has been removed. Afterwards, the remaining residue was dried at 30°C for 24 hours, and a total of 4.7 g of hemp seed meal was recovered.
상기 헴프씨드박 4.7g에 47㎖의 70%(v/v) 에탄올을 첨가하고 실온에서 180 rpm으로 3시간 동안 교반하여 추출한 후, 12,000 rpm으로 10분 동안 원심분리하여 얻은 상등액을 0.45㎛ 시린지 필터로 여과하여 헴프씨드박 추출물을 제조하였다. 47 ml of 70% (v/v) ethanol was added to 4.7 g of the hemp seed meal, extracted by stirring at room temperature at 180 rpm for 3 hours, and then centrifuged at 12,000 rpm for 10 minutes, and the obtained supernatant was filtered through a 0.45 ㎛ syringe filter. Hemp seed meal extract was prepared by filtration.
상기 헴프씨드박 추출물을 알루미늄 접시에 취하여 60℃에서 3시간, 85℃에서 30분, 90℃에서 30분 동안 건조시킨 후, 무게를 측정하였다. 이후, 하기식에 대입하여 추출 수율(%)을 계산하였다. The hemp seed meal extract was placed in an aluminum dish, dried at 60°C for 3 hours, 85°C for 30 minutes, and 90°C for 30 minutes, and then weighed. Afterwards, the extraction yield (%) was calculated by substituting the formula below.
추출 수율(%)={(건조 후 무게)/(추출 전 사용한 분말의 무게)}×100Extraction yield (%)={(weight after drying)/(weight of powder used before extraction)}×100
그 결과, 헴프씨드박 추출물의 추출 수율은 8.6%인 것을 확인하였다.As a result, it was confirmed that the extraction yield of the hemp seed meal extract was 8.6%.
실시예 1. 세포 독성 측정Example 1. Measurement of cytotoxicity
마우스 유래의 멜라노마 세포(B16F10)를 세포배양용 96웰 플레이트에 0.5×105 세포/㎖로 분주하여 24시간 동안 배양한 후, 50㎍/㎖, 100㎍/㎖ 및 200㎍/㎖의 헴프씨드박 추출물을 처리하고 24시간 동안 배양하였다. 이후, 5㎎/㎖의 MTT(3-(4,5-dimethylthiazole-2-yl)-2,5-biphenyl tetrazolium bromide) 시약을 각 웰에 넣고 3시간 배양 후, 상등액을 제거하고 100㎕의 DMSO(dimethyl sulfoxide)를 넣어 포르마잔(formazan) 결정체를 용해시키고, 마이크로플레이트 리더기를 이용하여 570 nm에서 흡광도를 측정하였다. 세포독성은 무처리군의 생존율을 100%로 하여 헴프씨드박 추출물 처리군의 상대적인 세포 생존율을 계산하였다. Mouse-derived melanoma cells (B16F10) were distributed at 0.5 The seed meal extract was treated and cultured for 24 hours. Afterwards, 5 mg/ml of MTT (3-(4,5-dimethylthiazole-2-yl)-2,5-biphenyl tetrazolium bromide) reagent was added to each well, and after 3 hours of incubation, the supernatant was removed and 100 ㎕ of DMSO was added. (dimethyl sulfoxide) was added to dissolve the formazan crystals, and the absorbance was measured at 570 nm using a microplate reader. For cytotoxicity, the relative cell survival rate of the hemp seed meal extract treated group was calculated by taking the survival rate of the untreated group as 100%.
그 결과, 50㎍/㎖, 100㎍/㎖ 및 200㎍/㎖의 헴프씨드박 추출물은 멜라노마 세포에서 세포 독성이 나타나지 않는 것을 확인하였다(도 1).As a result, it was confirmed that 50㎍/㎖, 100㎍/㎖ and 200㎍/㎖ hemp seed meal extract did not show cytotoxicity in melanoma cells (Figure 1).
실시예 2. 멜라닌 합성 억제능 측정Example 2. Measurement of melanin synthesis inhibition ability
마우스 유래의 멜라노마 세포(B16F10)를 12웰 플레이트에 0.5×105 세포/㎖로 분주하여 24시간 동안 배양한 후, 각 웰에 100nM의 α-MSH(melanocyte-stimulating hormone)를 처리하여 멜라닌 합성을 유도하였다. 이 때, 100㎍/㎖의 알부틴(arbutin) 또는 50㎍/㎖, 100㎍/㎖ 및 200㎍/㎖의 헴프씨드박 추출물을 동시에 처리하여 48시간 동안 배양하였다. 이후, 배지를 제거하고 PBS(phosphatate buffer saline)로 2회 세척하고, 각 웰에 용해 버퍼(0.1M 인산나트륨 버퍼, 1% triton X-100, 0.2mM PMSF)를 분주하여 -70℃에서 1시간 동안 방치하여 세포를 용해시켰다. 이후, 용해된 세포를 모아 원심분리하여 얻은 펠렛(pellet)에 10%(v/v) DMSO(dimethyl sulfoxide)가 첨가된 1N NaOH 용액 200㎕를 첨가하여 80℃에서 1시간 동안 반응시킨 후, 405 ㎚에서 흡광도를 측정하였다. 멜라닌 합성량은 α-MSH 단독 처리군 대비 백분율(%)로 표기하였다.Mouse-derived melanoma cells (B16F10) were distributed in a 12-well plate at 0.5 was derived. At this time, 100 μg/ml of arbutin or 50 μg/ml, 100 μg/ml, and 200 μg/ml of hemp seed meal extract were simultaneously treated and cultured for 48 hours. Afterwards, the medium was removed, washed twice with PBS (phosphatate buffer saline), and lysis buffer (0.1M sodium phosphate buffer, 1% triton The cells were lysed by leaving it for a while. Afterwards, 200 ㎕ of a 1N NaOH solution containing 10% (v/v) DMSO (dimethyl sulfoxide) was added to the pellet obtained by collecting the dissolved cells and centrifuging them, and reacted at 80°C for 1 hour, followed by 405 Absorbance was measured at nm. The amount of melanin synthesis was expressed as a percentage (%) compared to the α-MSH treatment group alone.
그 결과, 헴프씨드박 추출물은 α-MSH에 의해 증가된 멜라닌 합성량을 농도 의존적으로 감소시키는 것을 확인하였다(도 2). As a result, it was confirmed that the hemp seed meal extract reduced the amount of melanin synthesis increased by α-MSH in a concentration-dependent manner (Figure 2).
실시예 3. 티로시나아제 활성 억제능 측정Example 3. Measurement of tyrosinase activity inhibition ability
마우스 유래의 멜라노마 세포(B16F10)를 12웰 플레이트에 0.5×105 세포/㎖로 분주하여 24시간 동안 배양한 후, 각 웰에 100nM의 α-MSH(melanocyte-stimulating hormone)를 처리하여 티로시나아제 활성을 유도하였다. 이 때, 100㎍/㎖의 알부틴(arbutin) 또는 50㎍/㎖, 100㎍/㎖ 및 200㎍/㎖의 헴프씨드박 추출물을 동시에 처리하여 48시간 동안 배양하였다. 이후, 배지를 제거하고 PBS(phosphatate buffer saline)로 2회 세척하고, 각 웰에 용해 버퍼(0.1M 인산나트륨 버퍼, 1% triton X-100, 0.2mM PMSF)를 분주하여 -70℃에서 1시간 동안 방치하여 세포를 용해시켰다. 이후, 용해된 세포를 모아 원심분리하여 얻은 상등액을 얻었다. 0.1M 인산나트륨 버퍼(pH 6.8) 1㎖에 10mM L-3,4-디히드록시페닐알라닌(L-3,4-dihydroxyphenylalanine, L-DOPA)을 녹인 기질액 140㎕에 상기 상등액 60㎕를 혼합하여 37℃에서 1시간 반응시킨 후, 490 nm에서 흡광도를 측정하였다.Mouse-derived melanoma cells (B16F10) were dispensed in a 12-well plate at 0.5 The enzyme activity was induced. At this time, 100 μg/ml of arbutin or 50 μg/ml, 100 μg/ml, and 200 μg/ml of hemp seed meal extract were simultaneously treated and cultured for 48 hours. Afterwards, the medium was removed, washed twice with PBS (phosphatate buffer saline), and lysis buffer (0.1M sodium phosphate buffer, 1% triton The cells were lysed by leaving it for a while. Afterwards, the lysed cells were collected and centrifuged to obtain a supernatant. Mix 60㎕ of the supernatant with 140㎕ of substrate solution containing 10mM L-3,4-dihydroxyphenylalanine (L-DOPA) dissolved in 1㎖ of 0.1M sodium phosphate buffer (pH 6.8). After reacting at 37°C for 1 hour, absorbance was measured at 490 nm.
그 결과, 헴프씨드박 추출물은 α-MSH에 의해 증가된 티로시나아제의 활성을 농도 의존적으로 감소시키는 것을 확인하였다(도 3). As a result, it was confirmed that the hemp seed meal extract reduced the activity of tyrosinase increased by α-MSH in a concentration-dependent manner (Figure 3).
실시예 4. MITF 유전자 발현양 분석Example 4. MITF gene expression level analysis
티로시나아제 및 멜라닌 합성에 관여하는 TRP1(tyrosinase related protein 1) 단백질의 전사인자인 MITF(microphthalmia-associated transcription factor)의 mRNA 발현양을 분석하였다. The mRNA expression level of MITF (microphthalmia-associated transcription factor), a transcription factor for the TRP1 (tyrosinase related protein 1) protein involved in tyrosinase and melanin synthesis, was analyzed.
마우스 유래의 멜라노마 세포(B16F10)를 6웰 플레이트에 0.5×105 세포/㎖로 분주하여 24시간 동안 배양한 후, 각 웰에 100nM의 α-MSH(melanocyte-stimulating hormone)를 처리하여 티로시나아제 활성을 유도하였다. 이 때, 100㎍/㎖의 알부틴(arbutin) 또는 25㎍/㎖, 50㎍/㎖ 및 100㎍/㎖의 헴프씨드박 추출물을 동시에 처리하여 48시간 동안 배양하였다. 이후, 원심분리하여 상층액을 제거하고 얻은 세포 펠렛에 TRIzol시약을 처리하여 mRNA를 추출하고 정량한 후, cDNA 합성 키트를 이용하여 cDNA를 합성하였고, 하기 표 1과 같은 프라이머를 이용하여 PCR을 수행하였다. Mouse-derived melanoma cells (B16F10) were distributed in a 6-well plate at 0.5 The enzyme activity was induced. At this time, 100 μg/ml of arbutin or 25 μg/ml, 50 μg/ml, and 100 μg/ml of hemp seed meal extract were simultaneously treated and cultured for 48 hours. Afterwards, centrifugation was performed to remove the supernatant, and the obtained cell pellet was treated with TRIzol reagent to extract and quantify mRNA. cDNA was synthesized using a cDNA synthesis kit, and PCR was performed using primers as shown in Table 1 below. did.
그 결과, 헴프씨드박 추출물은 α-MSH에 의해 증가된 MITF 유전자의 발현을 농도 의존적으로 감소시키는 것을 확인하였다(도 4). As a result, it was confirmed that the hemp seed meal extract reduced the expression of the MITF gene increased by α-MSH in a concentration-dependent manner (Figure 4).
<110> NIKKYUHBIO andong national university industry-Academic cooperation Foundation <120> Composition for skin whitening comprising extract of hempseed meal as effective component <130> PN20448 <160> 2 <170> KoPatentIn 3.0 <210> 1 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 1 agcgtgtatt ttccccacag 20 <210> 2 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 2 tagctcctta atgcggtcgt 20 <110> NIKKYUHBIO andong national university industry-Academic cooperation Foundation <120> Composition for skin whitening comprising extract of hempseed meal as effective component <130> PN20448 <160> 2 <170> KoPatentIn 3.0 <210> 1 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 1 agcgtgtatt ttccccacag 20 <210> 2 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 2 tagctcctta atgcggtcgt 20
Claims (7)
(2) 상기 추출액을 원심분리하여 상등액인 오일 성분을 제거하고 건조시켜 헴프씨드박을 회수하는 단계; 및
(3) 상기 헴프씨드박에 에탄올을 첨가하고 실온에서 교반하여 추출한 후, 원심분리하여 얻은 상등액을 여과하여 헴프씨드박 추출물을 회수하는 단계;를 포함하는 방법으로 제조된 헴프씨드박 추출물을 유효성분으로 함유하는 피부 미백용 화장료 조성물.(1) homogenizing hemp seeds by adding hexane and extracting them by stirring at room temperature;
(2) recovering the hemp seed meal by centrifuging the extract to remove the oil component of the supernatant and drying it; and
(3) adding ethanol to the hemp seed meal, extracting it by stirring at room temperature, and filtering the supernatant obtained by centrifugation to recover the hemp seed meal extract; A cosmetic composition for skin whitening containing.
(2) 상기 추출액을 원심분리하여 상등액인 오일 성분을 제거하고 건조시켜 헴프씨드박을 회수하는 단계; 및
(3) 상기 헴프씨드박에 에탄올을 첨가하고 실온에서 교반하여 추출한 후, 원심분리하여 얻은 상등액을 여과하여 헴프씨드박 추출물을 회수하는 단계;를 포함하는 방법으로 제조된 헴프씨드박 추출물을 유효성분으로 함유하는 멜라닌 색소 과다 침착 질환의 예방 또는 개선용 건강기능식품 조성물.(1) homogenizing hemp seeds by adding hexane and extracting them by stirring at room temperature;
(2) recovering the hemp seed meal by centrifuging the extract to remove the oil component of the supernatant and drying it; and
(3) adding ethanol to the hemp seed meal, extracting it by stirring at room temperature, and filtering the supernatant obtained by centrifugation to recover the hemp seed meal extract; A health functional food composition for preventing or improving melanin hyperpigmentation disease.
(2) 상기 추출액을 원심분리하여 상등액인 오일 성분을 제거하고 건조시켜 헴프씨드박을 회수하는 단계; 및
(3) 상기 헴프씨드박에 에탄올을 첨가하고 실온에서 교반하여 추출한 후, 원심분리하여 얻은 상등액을 여과하여 헴프씨드박 추출물을 회수하는 단계;를 포함하는 방법으로 제조된 헴프씨드박 추출물을 유효성분으로 함유하는 멜라닌 색소 과다 침착 질환의 예방 또는 치료용 약학 조성물.(1) homogenizing hemp seeds by adding hexane and extracting them by stirring at room temperature;
(2) recovering the hemp seed meal by centrifuging the extract to remove the oil component of the supernatant and drying it; and
(3) adding ethanol to the hemp seed meal, extracting it by stirring at room temperature, and filtering the supernatant obtained by centrifugation to recover the hemp seed meal extract; A pharmaceutical composition for preventing or treating melanin hyperpigmentation disease containing.
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