KR20190005369A - Composition for preventing or improving skin wrinkle comprising c-phycocyanin as effective component - Google Patents
Composition for preventing or improving skin wrinkle comprising c-phycocyanin as effective component Download PDFInfo
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- KR20190005369A KR20190005369A KR1020170085909A KR20170085909A KR20190005369A KR 20190005369 A KR20190005369 A KR 20190005369A KR 1020170085909 A KR1020170085909 A KR 1020170085909A KR 20170085909 A KR20170085909 A KR 20170085909A KR 20190005369 A KR20190005369 A KR 20190005369A
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- lotion
- phycocyanin
- preventing
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- cosmetic composition
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Abstract
Description
본 발명은 씨-피코시아닌을 유효성분으로 함유하는 피부 주름의 예방 또는 개선용 조성물에 관한 것이다.The present invention relates to a composition for preventing or improving skin wrinkles containing cipicocyanin as an active ingredient.
피부는 외부환경의 자극으로부터 체내의 기관들을 보호해주며, 체온조절 등의 생체 항상성 유지에 중요한 역할을 한다. 이러한 피부는 여러 가지 내외적 요인에 의해서 노화가 발생하며, 크게 유전적 원인에 의한 내인성 노화와 환경적 원인에 의한 외인성 노화로 구분된다. 이 중, 광노화는 자외선(ultraviolet; UV)에 의한 노화를 의미한다. 최근 환경오염으로 인한 오존층 파괴는 자외선의 양을 증가시켰고 이에 따라 광노화에 대한 연구가 주목되고 있다(Wang SQ et al., 2010, Dermatol. Ther. 23(1):31-47). 광노화 된 피부에서는 거칠어짐, 탄력 손실, 주름 발생 및 불규칙한 색소 침착 등과 같은 외관상의 특징이 관찰되며, 이 중 광노화의 주된 연구 분야는 피부 주름의 변화에 대한 것이다. 상기 광노화에 의한 피부 주름 형성에 관해 피부의 주요 구성성분인 콜라겐(collagen)의 합성, 분해 및 수분 함유량 등의 기초적인 생리 대사 변화에 대한 연구 결과가 다수 보고되고 있다(Brenneisen et al., 2002, Ann. N. Y. Acad. Sci., 973:31-43). 특히 자외선에 의해 활성 산소종(reactive oxygen species) 생성이 증가되고 피부의 효소적, 비효소적 항산화 방어체계가 붕괴되어 콜라겐의 분해 증가 및 생합성을 감소시켜 진피층 내의 콜라겐이 현저하게 감소된다(Bickers DR, Athar M, 2006, J. Invest. Dermatol., 126(12):2565-2575). 상기 콜라겐 감소에 큰 영향을 미치는 것은 콜라겐 분해 효소(MMPs; matrix metalloproteinases)로, 이는 세포외기질(extracellular matrix)과 기저막(basement membrane)의 분해에 관여한다. 상기 효소는 자외선에 의해 활성이 증가되며 이를 억제함으로써 자외선에 의해 유도되는 피부 두께 증가 및 주름 형성이 감소된다는 연구 결과들이 보고되어 있다(Inomata S et al., 2003, J. Invest. Dermatol., 120(1):128-134). 따라서 광노화의 예방 및 치료를 위해서는 MMPs를 조절하는 것이 효과적인 방법으로 알려져 있다.Skin protects the organs in the body from external stimuli and plays an important role in the maintenance of body homeostasis, such as body temperature control. Such skin is aged by various internal and external factors and is divided into endogenous aging by genetic cause and exogenous aging by environmental cause. Among them, photoaging refers to aging by ultraviolet (UV). Recently, the destruction of the ozone layer due to environmental pollution has increased the amount of ultraviolet rays, and accordingly, research on photoaging has been attracting attention (Wang SQ et al., 2010, Dermatol. Ther. 23 (1): 31-47). In the photoaged skin, appearance characteristics such as roughness, loss of elasticity, wrinkles and irregular pigmentation are observed. Among them, the main research field of photoaging is the change of the skin wrinkles. There have been many reports on the basic physiological metabolism changes such as synthesis, degradation, and moisture content of collagen, which is a major constituent of skin, in the formation of skin wrinkles by photoaging (Brenneisen et al., 2002, Ann. NY Acad. Sci., 973: 31-43). Particularly, ultraviolet rays increase the production of reactive oxygen species, collapse of enzymatic and non-enzymatic antioxidant defenses of skin, decrease collagen decomposition and biosynthesis, and decrease collagen in the dermis (Bickers DR , Athar M, 2006, J. Invest. Dermatol., 126 (12): 2565-2575). Collagen degradation enzymes (MMPs) are a major contributor to collagen degradation, which is involved in the degradation of the extracellular matrix and basement membrane. Studies have been reported that the enzyme increases activity by ultraviolet rays and inhibits it, thereby increasing skin thickness and wrinkling induced by ultraviolet light (Inomata S et al., 2003, J. Invest. Dermatol., 120 (1): 128-134). Therefore, it is known that effective control of MMPs is required for prevention and treatment of photoaging.
한편, 씨-피코시아닌(c-phycocyanin)이라는 청색의 천연색소 단백질이 많이 함유된 것으로 알려져 있다. 씨-피코시아닌은 수용성이며 아주 강한 형광을 나타내고 항산화 기능이 매우 뛰어나, 식품, 의약품 및 화장품 원료로서 주목을 받고 있다. 일례로, 씨-피코시아닌은 담즙 색소와 같이 지방의 소화를 도와주고, 면역조절 기능, 항산화, 항암, 항혈액응고 및 항바이러스 효과 등을 의료에 이용하기 위한 연구가 이루어지고 있다. 또한 씨-피코시아닌은 염증을 유발하는 COX-2 효소의 억제제로서 알레르기성 염증 반응을 억제하는 것으로 알려져 있어, 아토피성 피부염과 같은 알레르기성 질환의 개선 및 치료 용도로도 연구되고 있다.On the other hand, it is known that c-phycocyanin contains a large amount of blue pigment. C-Pococyanin is water-soluble, has a very strong fluorescence and is highly antioxidant, and has attracted attention as a food, medicine and cosmetic raw material. For example, ciprocyanine has been studied for the purpose of helping the digestion of fat, such as bile pigment, and for immuno-controlling function, antioxidant, anti-cancer, anti-blood coagulation and antiviral effect. It is also known that ciprocycin inhibits the allergic inflammatory response as an inhibitor of the inflammatory COX-2 enzyme and has been studied for the improvement and treatment of allergic diseases such as atopic dermatitis.
씨-피코시아닌 관련 선행기술로는 한국공개특허 제2017-0036879호에 화장품 원료로 이용하기 위한 피코시아닌의 추출 및 안정화 방법이 개시되어 있고, 한국공개특허 제2015-0143914호에 스피루리나 추출물을 유효성분으로 함유하는 비만 예방 및 치료용 약학적 조성물이 개시되어 있으나, 본 발명의 씨-피코시아닌을 유효성분으로 함유하는 피부 주름의 예방 또는 개선용 조성물에 관한 선행기술은 개시된 바 없다.As a prior art related to cicapacyin, Korean Patent Laid-Open Publication No. 2017-0036879 discloses a method for extracting and stabilizing phycocyanin for use as a cosmetic raw material, and Korean Patent Laid-Open Publication No. 2015-0143914 discloses a method for extracting spirulina There is disclosed a pharmaceutical composition for preventing and treating obesity containing an active ingredient, but no prior art relating to a composition for preventing or improving skin wrinkles containing the present c-phycocyanin as an active ingredient has been disclosed.
본 발명은 상기와 같은 요구에 의해 도출된 것으로서, 본 발명은 씨-피코시아닌을 유효성분으로 함유하는 피부 주름의 예방 또는 개선용 조성물을 제공하고, 본 발명의 유효성분인 씨-피코시아닌(c-phycocyanin)은 세포 독성이 없으며, 자외선 조사에 의한 세포 손상으로부터 세포를 보호하고, 자외선 조사에 의해 증가된 MMP-1의 발현을 현저하게 억제하는 것을 확인함으로써, 본 발명을 완성하였다.The present invention provides a composition for prevention or improvement of skin wrinkles containing cipicocyanin as an active ingredient and a composition for preventing or improving wrinkles of skin, (c-phycocyanin) has no cytotoxicity, protects cells from damage by ultraviolet irradiation, and significantly inhibits the expression of MMP-1, which is increased by ultraviolet irradiation, thereby completing the present invention.
상기 목적을 달성하기 위하여, 본 발명은 씨-피코시아닌을 유효성분으로 함유하는 피부 주름의 예방 또는 개선용 화장료 조성물을 제공한다.In order to accomplish the above object, the present invention provides a cosmetic composition for preventing or improving skin wrinkles containing cipicocyanin as an active ingredient.
또한, 본 발명은 씨-피코시아닌을 유효성분으로 함유하는 피부 주름의 예방 또는 개선용 건강기능식품 조성물을 제공한다.The present invention also provides a health functional food composition for preventing or improving skin wrinkles containing cipicocyanin as an active ingredient.
또한, 본 발명은 씨-피코시아닌을 유효성분으로 함유하는 피부 주름의 예방 또는 치료용 약학 조성물을 제공한다.The present invention also provides a pharmaceutical composition for preventing or treating skin wrinkles containing cipicocyanin as an active ingredient.
본 발명은 씨-피코시아닌을 유효성분으로 함유하는 피부 주름의 예방 또는 개선용 조성물에 관한 것으로, 본 발명의 씨-피코시아닌이 자외선 조사에 의해 손상된 세포를 보호하며, 자외선 조사에 의해 증가된 MMP-1의 발현을 현저하게 억제하는 것이다. 따라서 본 발명의 화장료 조성물은 피부 주름의 예방 또는 개선용 화장료에 유용하게 사용될 수 있다. FIELD OF THE INVENTION The present invention relates to a composition for preventing or improving skin wrinkles containing cipicocyanin as an active ingredient. The present invention relates to a composition for protecting or repairing damaged skin cells by irradiation with ultraviolet rays, Gt; MMP-1 < / RTI > Therefore, the cosmetic composition of the present invention can be usefully used in cosmetics for preventing or improving skin wrinkles.
도 1은 본 발명의 씨-피코시아닌의 농도별(10, 50, 100, 200 및 250㎍/㎖) 처리에 따른 HaCaT 세포의 세포생존률(%)을 확인한 MTS 어세이 결과이다.
도 2는 UVB 조사에 따른 세포 손상에 대하여, 본 발명의 씨-피코시아닌의 HaCaT 세포의 세포 보호 효과를 확인한 것으로, HaCaT 세포(사람 각질형성 세포주)에 24시간 동안 각각의 시료를 농도별(10, 50, 100 및 200㎍/㎖)로 처리하고, 자외선(UV Crosslinker, UltR5 Lum)을 30분간 조사한 후, MTS 어세이를 수행한 결과이다. #은 UVB 조사군의 세포 생존률이 Control 군에 비해 통계적으로 유의미하게 감소했다는 것으로, p<0.05임을 의미하고, *은 씨-피코시아닌을 처리하고 UVB를 조사한 실험군의 세포 생존률이 UVB 조사군의 세포 생존률에 비교하여 통계적으로 유의미하게 증가하였다는 것으로, p<0.05임을 의미한다.
도 3은 본 발명의 씨-피코시아닌 처리에 따른 MMP-1의 발현량 변화를 확인한 결과이다. #은 UVB 조사군의 MMP-1 발현량이 Control 군에 비해 통계적으로 유의미하게 증가했다는 것으로, p<0.05임을 의미하고, *은 씨-피코시아닌을 처리하고 UVB를 조사한 실험군의 MMP-1 발현량이 UVB 조사군의 MMP-1 발현량에 비교하여 통계적으로 유의미하게 감소하였다는 것으로, p<0.05임을 의미한다.FIG. 1 shows MTS assay results showing the cell survival rate (%) of HaCaT cells according to the treatment of cicapicansin (10, 50, 100, 200 and 250 / / ml) according to the present invention.
FIG. 2 shows the cytoprotective effect of HaCaT cells of the present invention against HaCaT cells (human keratinocyte line) for 24 hours against the cell damage caused by UVB irradiation. (UV Crosslinker, UltR5 Lum) for 30 minutes, and then subjected to an MTS assay. # Indicates that the cell survival rate of the UVB irradiation group was statistically decreased compared with that of the control group, which means that the cell survival rate of the UVB-irradiated group treated with cipicocyanin was lower than that of the UVB irradiation group Which is statistically significant compared to the cell survival rate, which means that p <0.05.
FIG. 3 shows the results of confirming the change in the expression level of MMP-1 according to the cipicocyanin treatment of the present invention. # Indicates that the amount of MMP-1 expression in the UVB-irradiated group was statistically significantly increased compared to that in the control group, which means that p <0.05, and * indicates the amount of MMP-1 expression in the experimental group treated with cipicocyanin and UVB , Which is a statistically significant decrease compared to the amount of MMP-1 expression in the UVB-irradiated group, which means that p <0.05.
본 발명은 씨-피코시아닌(c-phycocyanin)을 유효성분으로 함유하는 피부 주름의 예방 또는 개선용 화장료 조성물에 관한 것이다. The present invention relates to a cosmetic composition for preventing or improving skin wrinkles containing c-phycocyanin as an active ingredient.
상기 씨-피코시아닌은 미세조류에서 유래한 것이 바람직하며, 더 바람직하게는 시아노박테리아 또는 홍조류로부터 유래한 것이지만 이에 한정하지 않는다. The cipicocyanin is preferably derived from microalgae, more preferably from cyanobacteria or red algae, but is not limited thereto.
상기 화장료 조성물은 MMP-1의 발현을 저해하는 것이 특징이다. The cosmetic composition is characterized by inhibiting the expression of MMP-1.
본 발명의 일 구현 예에 따른 화장료 조성물에 있어서, 상기 피부 주름의 예방 또는 개선용 화장료 조성물은 피부 외용 연고, 크림, 유연화장수, 영양화장수, 팩, 에센스, 헤어토닉, 샴푸, 린스, 헤어 컨디셔너, 헤어 트리트먼트, 젤, 스킨로션, 스킨소프너, 스킨토너, 아스트린젠트, 로션, 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 prevention or improvement of wrinkles of the skin may be used as an external ointment for skin, cream, softening longevity, nutritional lotion, pack, essence, hair tonic, shampoo, rinse, hair conditioner, Lotion, milky lotion, moisturizing lotion, nutrition lotion, massage cream, nutritional cream, moisturizing cream, hand cream, foundation, nutrition essence, sunscreen, soap, skin lotion, skin lotion, skin toner, astringent, lotion, But are not limited to, any one form selected from the group consisting of cleansing foams, cleansing lotions, cleansing creams, body lotions and body cleansers. The cosmetic composition comprising each of these formulations may contain various bases and additives necessary for formulation of the formulation, and the kinds and amounts of these ingredients can be easily selected by those 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, a cream or a gel, an animal fiber, a plant fiber, a wax, a paraffin, a starch, a tragacanth, a cellulose derivative, a polyethylene glycol, a silicone, a bentonite, Etc. may 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 추가로 클로로플루오로히드로카본, 프로판-부탄 또는 디메틸 에테르와 같은 추진체를 포함할 수 있다.When the formulation of the cosmetic composition of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as a carrier component, Propellants such as hydrocarbons, propane-butanes or dimethyl ethers.
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 an emulsion, a solvent, a solvent or an emulsifier is used as a carrier component, and examples thereof include water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, Glycol, 1,3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol or fatty acid esters of 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, a carrier such as water, a liquid diluent such as ethanol or propylene glycol, a suspension such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, Microcrystalline cellulose, aluminum metahydroxide, bentonite, agar or tracant, etc. 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, 리놀린 유도체 또는 에톡실화 글리세롤 지방산 에스테르 등이 이용될 수 있다.When the formulation of the cosmetic composition of the present invention is an interfacial-active agent-containing cleansing, the carrier component may include aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, acethionate, imidazolinium derivative, methyltaurate, sarcosinate , Fatty acid amide ether sulfate, alkylamidobetaine, aliphatic alcohol, fatty acid glyceride, fatty acid diethanolamide, vegetable oil, linolenic derivative or ethoxylated glycerol fatty acid ester.
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 an excipient including a fluorescent substance, a fungicide, a hygroscopic substance, a moisturizer, a fragrance, a fragrance carrier, a protein, a solubilizer, a sugar derivative, a sunscreen, a vitamin, .
또한, 본 발명은 씨-피코시아닌을 유효성분으로 함유하는 피부 주름의 예방 또는 개선용 건강기능식품 조성물에 관한 것이다.The present invention also relates to a health functional food composition for preventing or improving skin wrinkles containing cipicocyanin as an active ingredient.
본 발명의 건강기능식품 조성물은 씨-피코시아닌을 그대로 첨가하거나 다른 식품 또는 식품 성분과 함께 혼합하여 제조될 수 있고, 통상적인 방법에 따라 적절하게 제조될 수 있다. 상기 씨-피코시아닌을 첨가할 수 있는 식품의 예로는 카라멜, 육류, 소시지, 빵, 초콜릿, 캔디류, 스넥류, 과자류, 피자, 라면, 기타 면류, 껌류, 아이스크림류를 포함한 낙농제품, 각종 수프, 음료수, 차, 드링크제, 알코올 음료 및 비타민 복합제 중에서 선택된 어느 하나의 형태일 수 있으며, 통상적인 의미에서의 건강기능식품을 모두 포함한다. 즉, 상기 식품의 종류에는 특별한 제한은 없다. 상기 건강기능식품 조성물은 여러 가지 영양제, 비타민, 광물(전해질), 합성 및 천연 풍미제, 착색제 및 증진제(치즈, 초콜릿 등), 펙트산 및 그의 염, 알킨산 및 그의 염, 유기산, 보호성 콜로이드 증점제, pH 조절제, 안정화제, 방부제, 글리세린, 알코올, 탄산음료에 사용되는 탄산화제 등을 함유할 수 있다. 또한, 천연 과일 주스 및 야채 음료의 제조를 위한 과육을 함유할 수 있다. 상기의 성분은 독립적으로 또는 조합하여 사용할 수 있다. 또한, 본 발명의 건강기능식품 조성물은 여러 가지 향미제 또는 천연 탄수화물 등을 추가 성분으로서 함유할 수 있으며, 상기 천연 탄수화물은 포도당, 과당과 같은 단당류, 말토스, 슈크로스와 같은 이당류, 및 덱스트린, 사이클로덱스트린과 같은 다당류, 자일리톨, 소르비톨, 에리트리톨 등의 당알콜이다. 상기 천연 탄수화물의 비율은 크게 중요하지 않지만, 본 발명의 조성물 100g에 대하여, 0.01~0.04g 인 것이 바람직하고, 더욱 바람직하게는 0.02~0.03g을 포함하는 것이지만 이에 한정하지 않는다. 감미제로서는 타우마틴, 스테비아 추출물과 같은 천연 감미제나, 사카린, 아스파르탐과 같은 합성 감미제 등을 사용할 수 있다. The health functional food composition of the present invention can be prepared by directly adding cipicocyanin or mixing with other food or food ingredients, and can be suitably prepared according to a conventional method. Examples of foods that can be added with the above-mentioned cicapycia include dairy products including caramel, meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gums, ice cream, Beverage, tea, drink, alcoholic beverage, and vitamin complex, and includes all the health functional foods in the conventional sense. That is, there is no particular limitation on the kind of the food. The health functional food composition may contain various nutrients, vitamins, minerals (electrolytes), synthetic and natural flavors, colorants and enhancers (cheese, chocolate etc.), pectic acid and its salts, alkynic acid and its salts, Thickening agents, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonating agents used in carbonated drinks, and the like. It may also contain flesh for the production of natural fruit juices and vegetable drinks. The above components can be used independently or in combination. In addition, the health functional food composition of the present invention may contain various flavors or natural carbohydrates as an additional ingredient. The natural carbohydrates may be selected from the group consisting of glucose, monosaccharides such as fructose, disaccharides such as maltose and sucrose, Polysaccharides such as cyclodextrin, and sugar alcohols such as xylitol, sorbitol and erythritol. Although the ratio of the natural carbohydrate is not critical, it is preferably 0.01 to 0.04 g, more preferably 0.02 to 0.03 g, per 100 g of the composition of the present invention, but is not limited thereto. Examples of sweeteners include natural sweeteners such as tau martin and stevia extract, synthetic sweeteners such as saccharin and aspartame, and the like.
또한, 본 발명은 씨-피코시아닌을 유효성분으로 함유하는 피부 주름의 예방 또는 치료용 약학 조성물을 제공한다.The present invention also provides a pharmaceutical composition for preventing or treating skin wrinkles containing cipicocyanin as an active ingredient.
본 발명의 약학 조성물은 약학적으로 허용되는 담체, 부형제 또는 희석제를 추가로 포함할 수 있다. 본 발명의 약학 조성물에 포함되는 약학적으로 허용되는 담체는 제제 시에 통상적으로 이용되는 것으로서, 식염수, 멸균수, 링거액, 완충 식염수, 덱스트로즈 용액, 말토덱스트린 용액, 글리세롤, 에탄올, 락토스, 덱스트로스, 수크로스, 솔비톨, 만니톨, 전분, 아카시아 고무, 인산 칼슘, 알기네이트, 젤라틴, 규산 칼슘, 미세결정성 셀룰로스, 폴리비닐피롤리돈, 셀룰로스, 시럽, 메틸 셀룰로스, 메틸히드록시벤조에이트, 프로필히드록시벤조에이트, 활석, 스테아르산 마그네슘 및 미네랄 오일 등을 포함하나, 이에 한정되는 것은 아니다. 또한 상기 성분들 이외에 항산화제, 완충액, 정균제, 희석제, 계면활성제, 결합제, 윤활제, 습윤제, 감미제, 향미제, 유화제, 현탁제 또는 보존제 등을 추가로 포함할 수 있다. 본 발명의 약학 조성물은 경구 또는 비경구 투여할 수 있으며, 비경구 투여는 주사 또는 피부에 도포하는 방법으로 투여할 수 있는 것이다. 본 발명의 약학 조성물의 적합한 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 pharmaceutical composition of the present invention may further comprise a pharmaceutically acceptable carrier, excipient or diluent. The pharmacologically acceptable carrier to be contained in the pharmaceutical composition of the present invention is one which is usually used at the time of formulation and includes saline, sterilized water, Ringer's solution, buffered saline, dextrose solution, maltodextrin solution, glycerol, ethanol, But are not limited to, sucrose, sucrose, sucrose, sorbitol, mannitol, starch, acacia, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, syrup, methylcellulose, methylhydroxybenzoate, Hydroxybenzoate, talc, magnesium stearate and mineral oil, and the like. In addition, the composition may further contain an antioxidant, a buffer, a bacteriostatic agent, a diluent, a surfactant, a binder, a lubricant, a wetting agent, a sweetener, a flavoring agent, an emulsifier, a suspending agent or a preservative. The pharmaceutical composition of the present invention can be administered orally or parenterally, and parenteral administration can be carried out by injection or application to skin. A suitable dose of the pharmaceutical composition of the present invention may be variously prescribed by factors such as the formulation method, the administration method, the age, body weight, sex, pathological condition, food, administration time, administration route, excretion rate and responsiveness of the patient .
이하, 실시예를 이용하여 본 발명을 더욱 상세하게 설명하고자 한다. 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 described in more detail with reference to Examples. It is to be understood by those skilled in the art that these embodiments are merely illustrative of the present invention and that the scope of the present invention is not limited thereto.
1. 씨-피코시아닌의 준비1. Mr. - Preparation of Pycocyanin
본 발명의 일실시예에서 사용한 씨-피코시아닌은 시그마알드리치(주)로부터 구입하였고, 정제없이 사용하였다. The cipicocyanin used in one embodiment of the present invention was purchased from Sigma-Aldrich and used without purification.
2. 세포배양2. Cell culture
세포 생존율, 세포보호 및 MMP-1의 발현량 분석은 HaCaT 세포를 이용하였으며, HaCaT 세포는 Dulbecco's Modified Eagle Medium (DMEM, Gibco)에 10% 소태아 혈청(Fetal bovine serum;FBS, Gibco)과 1% 페니실린-스트렙토마이신(penicillin-streptomycin;PS, Gibco)이 함유된 배지에서 37℃, 5% CO2 조건에서 배양하였다.HaCaT cells were cultured in Dulbecco's Modified Eagle Medium (DMEM, Gibco) supplemented with 10% fetal bovine serum (FBS, Gibco) and 1% And cultured in a medium containing penicillin-streptomycin (PS, Gibco) at 37 ° C and 5% CO 2 .
실시예Example 1. One. 세포생존률Cell survival rate 분석 analysis
본 발명의 유효성분인 씨-피코시아닌을 농도별(10, 50, 100, 200 및 250㎍/㎖)로 희석하여, 상기 배양한 HaCaT 세포에 24시간 동안 처리하고, MTS 어세이(CellTiter Aqueous One Solution Cell proliferation assay kit, 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophynyl)-2H-tetrazolium, inner salt; MTS, Promega Co. Madison, WI, USA) 기법으로 세포 생존율을 490nm에서 마이크로플레이트 리더(Molecular Devices, Sunnyvale, CA, USA)로 측정하였다. The cultured HaCaT cells were treated for 24 hours, and the MTS assay (CellTiter Aqueous) was applied to the cultured HaCaT cells by diluting the active ingredient of the present invention, cicapycyanine, at different concentrations (10, 50, 100, 200 and 250 / / One Solution Cell proliferation assay kit was prepared by adding 3- (4,5-dimethylthiazol-2-yl) -5- (3-carboxymethoxyphenyl) -2- (4-sulfophynyl) -2H-tetrazolium, inner salt MTS, Promega Co. Madison , WI, USA), the cell viability was measured at 490 nm using a microplate reader (Molecular Devices, Sunnyvale, Calif., USA).
그 결과, 도 1에 개시한 바와 같이 본 발명의 시료를 처리하였을 때의 상대적 세포생존률(%)이 다소 증가하였거나 유지하였다. 따라서, 본 발명의 씨-피코시아닌은 세포 독성이 거의 없다는 것을 알 수 있었다.As a result, the relative cell viability (%) when the sample of the present invention was treated was slightly increased or maintained as shown in Fig. Therefore, it was found that the present c-phycocyanin had little cytotoxicity.
실시예Example 2. 자외선 조사에 따른 세포 보호 효과 확인 2. Confirmation of cytoprotective effect by ultraviolet irradiation
HaCaT 세포에 24시간 동안 각각의 시료를 농도별로 희석하여 처리하고 자외선(UV Crosslinker, Ultra Lum)을 30분간 조사한 후, MTS 어세이(CellTiter Aqueous One Solution Cell proliferation assay kit, 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophynyl)-2H-tetrazolium, inner salt; MTS, Promega Co. Madison, WI, USA) 기법으로 세포 생존능을 490nm에서 마이크로플레이트 리더(Molecular Devices, Sunnyvale, CA, USA) 로 측정하여 시험하였다. After incubation for 30 min with ultraviolet light (UV Crosslinker, Ultra Lum), HaCaT cells were treated with dilution of each sample for 24 hours. Cells were stained with CellTiter Aqueous One Solution Cell proliferation assay kit, 3- (4,5- The cell viability was measured at 490 nm using a microplate reader (MTS, Promega Co., Madison, Wis., USA) Reader (Molecular Devices, Sunnyvale, Calif., USA).
그 결과, 도 2에 개시한 바와 같이 UVB 조사에 의해 세포생존률(%)이 약 50% 정도 감소한 것을 확인할 수 있었고, UV 조사 후, 본 발명의 씨-피코시아닌을 처리한 경우 세포생존률(%)이 농도 의존적으로 증가하는 것을 확인하여 본 발명의 씨-피코시아닌이 세포보호 효과가 있다고 판단하였다.As a result, it was confirmed that the cell viability (%) was reduced by about 50% by UVB irradiation as shown in FIG. 2, and the cell survival rate (%) after treatment with UV- ) Was increased in a concentration-dependent manner, indicating that the cell-protective effect of the present invention was demonstrated.
실시예Example 3. 3. MMPMMP -1의 발현량 분석-1 expression level
HaCaT 세포에 24시간 동안 각각의 시료를 처리하고 자외선(UV Crosslinker, Ultra Lum)을 20mJ/cm2로 조사한 후, MMP-1 ELISA kit(R&D Systems, Inc., Minneapolis, MN, USA)를 이용하여 MMP-1의 발현량을 측정하였다. Each sample was treated with HaCaT cells for 24 hours and irradiated with ultraviolet light (UV Crosslinker, Ultra Lum) at 20 mJ / cm 2. Then , using an MMP-1 ELISA kit (R & D Systems, Inc., Minneapolis, The expression level of MMP-1 was measured.
상세하게는 96웰 플레이트에 HaCaT 세포를 도말한 후, 본 발명의 씨-피코시아닌을 각각 농도별(10, 50, 100 및 200㎍/㎖)로 처리하였다. 이후, 자외선(UV) 조사하고, 배양물의 상등액을 수집하고 13,000rpm으로 5분 동안 원심분리하였다. 이후 상등액을 취해 ELISA 키트를 이용한 형광 어세이를 수행하여 MMP-1의 발현량을 정량분석하였다.Specifically, HaCaT cells were plated on a 96-well plate, and then the seed-phycocyanin of the present invention was treated with concentrations (10, 50, 100 and 200 μg / ml), respectively. Then, ultraviolet (UV) irradiation was performed, and the supernatant of the culture was collected and centrifuged at 13,000 rpm for 5 minutes. The supernatant was then subjected to fluorescent assay using an ELISA kit to quantitatively analyze the expression level of MMP-1.
그 결과, 도 3 에 개시한 바와 같이 자외선을 조사하지 않은 음성대조군에 비해, 자외선을 조사한 군에서 현저하게 MMP-1의 발현량이 증가한다는 것을 확인하였고, 본 발명의 씨-피코시아닌의 처리군에서 MMP-1의 발현량이 자외선 조사군에 비해 현저하게 농도의존적으로 감소하는 것을 확인하였다. As a result, it was confirmed that the expression level of MMP-1 was remarkably increased in the group irradiated with ultraviolet light, as compared with the negative control group not irradiated with ultraviolet light as shown in Fig. 3. In the group treated with cipicocyanine of the present invention , The expression level of MMP-1 was significantly decreased in a concentration-dependent manner as compared with the ultraviolet irradiation group.
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| CN115003269A (en) * | 2020-01-31 | 2022-09-02 | 株式会社资生堂 | Maintenance or promotion of collagen production ability |
| CN115023216A (en) * | 2020-01-31 | 2022-09-06 | 株式会社资生堂 | Antioxidant or skin stress inhibitor |
| CN115023213A (en) * | 2020-01-31 | 2022-09-06 | 株式会社资生堂 | Inflammation inhibitor |
| CN115038424A (en) * | 2020-01-31 | 2022-09-09 | 株式会社资生堂 | Photoaging inhibitor |
| CN115052579A (en) * | 2020-01-31 | 2022-09-13 | 株式会社资生堂 | Hyaluronic acid production promoter |
| WO2021153773A1 (en) * | 2020-01-31 | 2021-08-05 | 株式会社 資生堂 | Photoaging inhibitor |
| CN114557907A (en) * | 2022-03-14 | 2022-05-31 | 江苏大学 | Application of phycocyanin nanoparticles in preparation of sunscreen preparation |
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