KR102628074B1 - skin external application composition for anti-aging containing Luzula capitate extract - Google Patents
skin external application composition for anti-aging containing Luzula capitate extract Download PDFInfo
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- KR102628074B1 KR102628074B1 KR1020180063394A KR20180063394A KR102628074B1 KR 102628074 B1 KR102628074 B1 KR 102628074B1 KR 1020180063394 A KR1020180063394 A KR 1020180063394A KR 20180063394 A KR20180063394 A KR 20180063394A KR 102628074 B1 KR102628074 B1 KR 102628074B1
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- South Korea
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- skin
- skin wrinkles
- glycation
- pheasant
- extract
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Classifications
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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
- A61K8/96—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]
- A61K8/9794—Liliopsida [monocotyledons]
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/88—Liliopsida (monocotyledons)
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
- A61Q19/08—Anti-ageing preparations
Abstract
본 발명은 피부의 콜라겐 당화를 억제하여 피부 내 콜라겐 분해 저해를 통한 주름 개선과 피부의 투명도 개선에 도움을 줄 수 있는 꿩의밥 추출물을 유효성분으로 포함하는 항노화용 피부외용제 조성물을 제공한다. 상기 조성물은 항산화 효과가 우수하고, 당화반응(glycation)에 의한 콜라겐 당화를 억제하여 피부의 주름 생성을 예방하거나 개선할 수 있으며, 최종당화산물(advanced glycation end products, AGEs)의 생성을 억제하여 피부 단백질 당화에 의한 피부 투명도가 저하되는 것을 개선할 수 있다. 따라서 본 발명의 조성물은 피부노화 방지, 주름 개선, 피부 톤 개선 및 피부모공 축소 등을 위한 의약품, 화장품, 퍼스널 케어 제품 등으로 활용될 수 있다.The present invention provides an anti-aging external skin preparation composition containing pheasant rice extract as an active ingredient, which can help improve wrinkles and skin transparency by inhibiting collagen decomposition in the skin by inhibiting collagen glycation in the skin. The composition has excellent antioxidant effect, can prevent or improve the formation of wrinkles on the skin by inhibiting collagen glycation due to glycation, and inhibits the production of advanced glycation end products (AGEs), thereby protecting the skin. It can improve the decrease in skin transparency caused by protein glycation. Therefore, the composition of the present invention can be used as medicine, cosmetics, personal care products, etc. to prevent skin aging, improve wrinkles, improve skin tone, and reduce skin pores.
Description
본 발명은 꿩의밥 추출물을 유효성분으로 포함하는 항노화용 피부 외용제 조성물에 관한 것이다.The present invention relates to an external skin composition for anti-aging containing pheasant rice extract as an active ingredient.
피부의 노화는 나이가 들어가면서 자연적으로 발생하는 변화이며 그 밖에도 자외선, 담배, 식이습관 및 환경오염 등 많은 외부적 인자에 의해서 발생한다. 피부의 노화에는 겉보기 나이의 저하, 살결의 저하, 주름증가, 칙칙함의 증가, 피부기능의 저하 등이 있다. 피부는 나이 듦에 따라 붉은색이 감소, 노란색이 증가, 명도가 낮아지는 색조로 변화하여 피부가 칙칙한 상태로 된다. 칙칙함의 요인에는 피부의 색소침착, 단백질의 당화나 산화에 의한 착색 등이 관여한다.Skin aging is a natural change that occurs as we age, and is also caused by many external factors such as ultraviolet rays, cigarettes, dietary habits, and environmental pollution. Skin aging includes a decrease in apparent age, decrease in skin texture, increase in wrinkles, increase in dullness, and decrease in skin function. As the skin ages, the red color decreases, the yellow color increases, and the color tone changes to lower brightness, making the skin dull. Factors contributing to dullness include pigmentation of the skin and discoloration due to glycation or oxidation of proteins.
현재, 단백질 당화 억제제로 아미노구아니딘이 친핵성 히드라진(hydrazine)으로 아마도리 산물과 결합하여 단백질과의 교차결합을 방지함으로써 최종당화산물의 생성을 억제하여 가장 유망한 당화 억제제 의약품으로 제 3상 임상실험까지 진행되었으나, 장기간 투여 시 카탈라아제의 저해와 관련된 부작용이 우려되고 있어 안전하고 효능이 우수한 새로운 단백질 당화 억제 소재를 탐색하고자 하는 노력이 계속되어 왔다. 또한 체내 생성된 활성산소는 생체 단백질을 변성시키며, 변성된 생체 단백질은 당과의 반응성이 높아져 당화반응이 유도된다. 이러한 메커니즘에 따라 활성산소로 유도된 산화스트레스 억제능이 우수하고 페놀성 화합물이 풍부한 추출물의 항당화 연구가 최근까지도 활발히 진행되고 있지만 아직 실질적 효능을 구현하기에는 부족한 수준이다.Currently, aminoguanidine, a protein glycation inhibitor, binds to Amadori products with nucleophilic hydrazine and prevents cross-linking with proteins, thereby suppressing the production of advanced glycation products, making it the most promising glycation inhibitor drug in phase 3 clinical trials. However, there are concerns about side effects related to inhibition of catalase during long-term administration, so efforts have been made to explore new, safe and effective materials for inhibiting protein glycation. In addition, active oxygen generated in the body denatures biological proteins, and the denatured biological proteins become more reactive with sugars, leading to glycation reactions. According to this mechanism, anti-glycation research on extracts rich in phenolic compounds, which have an excellent ability to suppress oxidative stress induced by free radicals, has been actively conducted until recently, but are still insufficient to realize practical efficacy.
상기 문제점을 해결하기 위한 본 발명의 목적은 피부노화 방지, 피부주름 방지, 피부주름 개선, 피부 톤 개선 및 피부모공 축소를 상승시키고, 인체에 안전하며 장기간 사용하여도 부작용이 없는 항노화용 피부외용제 조성물을 제공하는 것이다.The purpose of the present invention to solve the above problems is to provide an anti-aging external skin preparation composition that prevents skin aging, prevents skin wrinkles, improves skin wrinkles, improves skin tone and increases skin pore reduction, is safe for the human body and has no side effects even when used for a long period of time. is to provide.
상기 목적을 달성하기 위한 본 발명의 양상은 꿩의밥 추출물을 유효성분으로 포함하는 항노화용 피부외용제 조성물을 제공하는 것이다.An aspect of the present invention to achieve the above object is to provide an external skin preparation composition for anti-aging containing pheasant rice extract as an active ingredient.
본 발명에 있어서, 상기 꿩의밥 추출물은 우수한 항산화 효과를 나타내며, 콜라겐 당화반응(collagen glycation)와 최종당화산물(advanced glycation end products, AGEs)의 생성을 억제시켜 피부노화 방지, 주름 개선, 피부 톤 개선 및 피부모공 축소에 효과적이다.In the present invention, the pheasant rice extract exhibits excellent antioxidant effects and inhibits collagen glycation and the production of advanced glycation end products (AGEs) to prevent skin aging, improve wrinkles, and improve skin tone. It is effective in improving and reducing skin pores.
또한, 피부 세포에 악영향을 미치지 않은 안전성을 가지면서 피부 세포 노화를 방지하므로 피부노화 방지, 주름 개선, 피부 톤 개선 및 피부모공 축소 등을 위한 의약품, 화장품, 퍼스널 케어 제품 등으로 활용될 수 있다.In addition, since it is safe and does not adversely affect skin cells and prevents skin cell aging, it can be used as medicine, cosmetics, and personal care products to prevent skin aging, improve wrinkles, improve skin tone, and reduce skin pores.
도 1은 피부 섬유아세포에 대한 꿩의밥 추출물의 농도 별 세포 생존율을 나타낸 그래프이다.
도 2는 꿩의밥 추출물과 아미노구아니딘 염산염의 섬유아세포 내 카복시메칠라이신(carboxymethyl lysine, CML) 생성 억제 정도를 나타낸 그래프이다.Figure 1 is a graph showing the cell survival rate by concentration of pheasant rice extract for skin fibroblasts.
Figure 2 is a graph showing the degree of inhibition of carboxymethyl lysine (CML) production in fibroblasts by pheasant rice extract and aminoguanidine hydrochloride.
이하 본 발명을 보다 상세히 설명한다. 다만, 본 발명의 설명을 위한 것으로, 본 발명의 범위는 청구범위에 기재된 사항을 기초로 한다.Hereinafter, the present invention will be described in more detail. However, this is for explanation of the present invention, and the scope of the present invention is based on the matters stated in the claims.
본 발명은 꿩의밥 추출물을 유효성분으로 포함하는 항노화용 피부외용제 조성물에 관한 것이다.The present invention relates to an external skin preparation composition for anti-aging containing pheasant rice extract as an active ingredient.
상기 꿩의밥 추출물은 상기 꿩의밥의 잎, 꽃, 열매, 줄기, 뿌리 및 종자로 이루어진 군으로부터 선택된 1종 이상의 추출물일 수 있으며, 전초를 사용할 수 있다.The pheasant rice extract may be one or more extracts selected from the group consisting of leaves, flowers, fruits, stems, roots, and seeds of the pheasant rice, and whole plant may be used.
상기 꿩의밥 추출물은 항산화력이 우수하고, 콜라겐 당화를 감소시켜 효과적으로 최종당화산물의 생성을 억제하여 피부노화 방지, 피부주름 방지, 피부주름 개선, 피부 톤 개선 및 피부모공 축소에 유용하게 사용될 수 있다.The pheasant rice extract has excellent antioxidant power and reduces collagen glycation, effectively suppressing the production of advanced glycation products, so it can be usefully used to prevent skin aging, prevent skin wrinkles, improve skin wrinkles, improve skin tone, and reduce skin pores. there is.
본 발명에 있어서, 상기 ‘항노화’란 피부보호, 피부상태 개선, 피부미백, 피부노화 및 주름의 예방 또는 개선, 피부모공 축소, 피부보호, 피부의 염증 반응 완화, 면역질환 개선능, 피부 장벽기능 개선, 피부자극 완화, 피부세포 증진, 재생능, 항산화능, 콜라겐 합성 증진능 등을 모두 포함하는 개념이다.In the present invention, the term 'anti-aging' refers to skin protection, improvement of skin condition, skin whitening, prevention or improvement of skin aging and wrinkles, reduction of skin pores, skin protection, alleviation of inflammatory reactions in the skin, ability to improve immune diseases, and skin barrier. It is a concept that includes functional improvement, skin irritation relief, skin cell enhancement, regenerative ability, antioxidant ability, and collagen synthesis enhancement ability.
또한, 상기 ‘유효성분으로 포함’의 의미는 피부외용제 조성물로써 항노화에 따른 피부 개선 효과를 나타낼 수 있는 예컨대 피부외용제 조성물로써 주름개선 효능과 관련된 콜라겐 합성, 멜라닌 형성(melanogenesis) 저해에 따른 피부미백 효능, 피부보습, 모공축소 또는 유도형 산화질소 합성 효소(inducible nitric oxide synthase, iNOS) 억제 활성, 피지억제, 항균력 등을 나타낼 수 있는 정도의 유효량을 함유하는 것을 의미한다.In addition, the meaning of 'included as an active ingredient' is a composition for external application for skin that can exhibit anti-aging skin improvement effects, such as collagen synthesis related to wrinkle improvement efficacy, and skin whitening due to inhibition of melanogenesis. It means that it contains an effective amount that can exhibit efficacy, skin moisturization, pore reduction, inducible nitric oxide synthase (iNOS) inhibitory activity, sebum inhibition, antibacterial activity, etc.
본 발명에 있어서, 상기 항노화용 피부외용제 조성물의 유효성분인 꿩의밥은 다음과 같은 특징을 가지고 있다.In the present invention, pheasant rice, which is an active ingredient of the anti-aging external skin preparation composition, has the following characteristics.
상기 꿩의밥(Luzula capitate)은 골풀목 골풀과의 여러해살이풀로 산기슭의 양지초원에 자생하며 동아시아의 온대와 난대에 분포한다. 줄기는 덩이 모양의 땅속줄기에서 뭉쳐나며 곧게 서고 높이가 10~30 cm이다. 꽃은 4~5월에 줄기 끝에 작은 꽃들이 뭉쳐 피어 둥근 공 모양의 두상꽃차례를 이룬다. 열매는 삭과이고 길이 2.5 mm인 모난 달걀 모양이며, 종자는 둥글거나 넓은 달걀 모양이며 검은 빛을 띠는 짙은 갈색이다. 꿩의밥의 지상부를 약으로 쓰며, 지양매라고 부른다. 적백리(곱과 피고름이 섞인 대변을 보는 이질)을 치료하는데 효능이 있다.The above-mentioned pheasant rice (Luzula capitate) is a perennial herb of the rush family and grows naturally in sunny meadows at the foot of mountains and is distributed in the temperate and subtropical regions of East Asia. The stems grow from tuber-shaped underground stems and stand upright, reaching a height of 10 to 30 cm. Flowers bloom in clusters of small flowers at the ends of stems in April and May, forming round, ball-shaped capitate inflorescences. The fruit is a capsule, angular and egg-shaped, 2.5 mm long, and the seeds are round or wide egg-shaped and dark brown with a black tint. The above-ground part of pheasant rice is used as medicine and is called Jiyangmae. It is effective in treating jeokbaekri (dysentery with feces mixed with mucus and blood).
본 발명의 일 실시예에 있어서, 상기 꿩의밥 추출물은 피부 콜라겐 당화반응(collagen glycation)과 최종당화산물(advanced glycation end products, AGEs)의 생성을 억제시킬 수 있다.In one embodiment of the present invention, the pheasant meal extract can inhibit skin collagen glycation and the production of advanced glycation end products (AGEs).
피부 노화의 주원인 중 하나인 당화반응(glycation)이란 단백질의 유리 아미노기인 리신(lysine)이나 아르기닌(arginine)과 환원당의 카보닐(carbonyl)기가 반응하여 초기 당화(glycation) 생성물인 시프 염기(schiff base)를 형성한다. 이 반응은 당류 농도에 의존적으로 진행되며, 수 시간 내 반응이 이루어진다. 생성된 시프 염기(schiff base)는 화학적 재배열(chemical rearrangement)을 통해 안정된 아마도리 산물(amadori product)을 생성하며, 수 일 내 반응이 이루어진다. 아마도리 산물이 체내 축적되어 일련의 산화, 탈수, 축합, 재전위, 분열, 고리화 등의 반응을 통해 단백질과 가교결합을 형성하는데 이 반응은 비가역적으로 이루어진다. 이러한 반응이 수 주에서 수 개월 정도 이루어지면 최종적으로 생성된 갈색 화합물의 당화산물인 최종당화산물(advanced glycation end products, AGEs)을 형성하는데 이 물질은 매우 안정하여 체내 축적될 경우 세포 내 외부에 이상을 초래한다.Glycation, one of the main causes of skin aging, is a reaction between lysine or arginine, a free amino group of a protein, and the carbonyl group of a reducing sugar, resulting in an initial glycation product, Schiff base. ) to form. This reaction proceeds depending on the saccharide concentration and occurs within a few hours. The resulting Schiff base generates a stable Amadori product through chemical rearrangement, and the reaction occurs within a few days. Amadori products accumulate in the body and form cross-links with proteins through a series of reactions such as oxidation, dehydration, condensation, retranslocation, cleavage, and cyclization, and this reaction occurs irreversibly. When this reaction takes place over several weeks to several months, advanced glycation end products (AGEs), which are the final glycation products of brown compounds, are formed. These substances are very stable and can cause abnormalities inside and outside of cells when accumulated in the body. causes
또한, 피부 내 생성된 갈색화합물의 최종당화산물(AGEs)이 축적될 경우 피부의 황색증가 및 얼룩을 유발하고, 멜라닌 생성을 촉진시키는 등 겉보기 노화에 관여하기 때문에 피부 항노화 대책으로 당화스트레스의 억제(항당화)를 의식하는 것이 중요하다. 당화반응은 고혈당을 가지는 당뇨환자에서 정상인보다 급격히 일어나지만 정상 혈당의 경우에도 노화가 진행됨에 따라 체내에서 일어나며, 구체적인 당화 대책으로는 식후 고혈당을 억제하고 당화반응을 억제하며 생성한 AGEs를 분해 배설하는 것 등이 있다.In addition, when advanced glycation end products (AGEs) of brown compounds generated in the skin accumulate, they cause increased yellowness and staining of the skin and are involved in apparent aging by promoting melanin production, so it is recommended to suppress glycation stress as a skin anti-aging measure. It is important to be conscious of (anti-glycosylation). The glycation reaction occurs more rapidly in diabetic patients with high blood sugar levels than in normal people, but even in normal blood sugar levels, it occurs in the body as aging progresses. Specific glycation measures include suppressing postprandial hyperglycemia, suppressing the glycation reaction, and breaking down and excreting the generated AGEs. There are things, etc.
또한, 최종당화산물(AGEs) 안에 가장 풍부하게 있는 카복시메칠라이신(carboxymethyl lysine, CML)은 당화 초기생성물의 산화적 분해반응에 의해 발생한다. I 형 콜라겐은 진피 안에서 가장 많은 단백질로 당화의 아주 좋은 표적이 된다. 당화스트레스로 인한 피부 콜라겐의 당화 발생시 콜라겐 단백질 변성 및 진피 내 CML 축적, 그리고 이미 생성된 최종당화산물(AGEs)과 가교결합 등의 반응이 지속됨에 따라 MMP-1(matrix metalloproteinase) 단백질 발현을 높게 유지시켜 콜라겐 분해를 촉진하여 피부의 유연성 및 탄력성이 감소되어 주름이 생성된다.In addition, carboxymethyl lysine (CML), which is most abundant in advanced glycation end products (AGEs), is generated by oxidative decomposition reaction of early glycation products. Type I collagen is the most abundant protein in the dermis and is a good target for glycation. When glycation of skin collagen occurs due to glycation stress, reactions such as collagen protein denaturation, CML accumulation in the dermis, and cross-linking with advanced glycation end products (AGEs) that have already been formed continue, maintaining high MMP-1 (matrix metalloproteinase) protein expression. This promotes the breakdown of collagen, reducing the flexibility and elasticity of the skin, resulting in the formation of wrinkles.
따라서, 상기 꿩의밥 추출물은 피부 콜라겐 당화반응(collagen glycation)과 최종당화산물(advanced glycation end products, AGEs)의 생성을 억제시켜 피부노화 방지, 피부주름 방지, 피부주름 개선, 피부 톤 개선 및 피부모공 축소의 효과가 뛰어나다.Therefore, the pheasant rice extract inhibits skin collagen glycation and the production of advanced glycation end products (AGEs), thereby preventing skin aging, preventing skin wrinkles, improving skin wrinkles, improving skin tone, and improving skin tone. It is highly effective in reducing pores.
상기 꿩의밥 추출물은 상기 피부외용제 조성물 전체 중량에 대하여 0.0001 내지 30중량%로 포함될 수 있으며, 바람직하게는 0.001 내지 10중량%이고, 보다 바람직하게는 0.1 내지 5중량%이다.The pheasant rice extract may be included in an amount of 0.0001 to 30% by weight, preferably 0.001 to 10% by weight, and more preferably 0.1 to 5% by weight, based on the total weight of the external skin composition.
상기 꿩의밥 추출물의 함량이 0.0001중량% 미만이면 유효한 효과를 나타낼 수 없으며, 30중량%를 초과하면 변색, 변취 등 제형 안정성에 문제가 발생될 수 있다.If the content of the pheasant rice extract is less than 0.0001% by weight, it cannot exhibit an effective effect, and if it exceeds 30% by weight, problems with formulation stability such as discoloration and off-odor may occur.
본 발명의 일 실시예에 있어서, 상기 꿩의밥 추출물은 물, 유기용매 또는 이들의 혼합물을 사용하여 당업계에 잘 알려진 통상적인 추출 방법에 의하여 추출할 수 있으며, 유기용매를 사용하여 추출하는 것이 바람직하다.In one embodiment of the present invention, the pheasant rice extract can be extracted by a conventional extraction method well known in the art using water, an organic solvent, or a mixture thereof, and extraction using an organic solvent is desirable.
상기 유기용매는 탄소수 1 내지 10의 무수 또는 함유 메탄올, 에탄올, 프로필 알코올, 부틸알코올, 글리세린, 프로필렌글리콜 및 부틸렌글리콜로 이루어진 군으로부터 선택되는 1종 이상일 수 있다.The organic solvent may be at least one selected from the group consisting of anhydrous or containing methanol, ethanol, propyl alcohol, butyl alcohol, glycerin, propylene glycol, and butylene glycol having 1 to 10 carbon atoms.
본 발명에 있어서, 상기 꿩의밥 줄기 추출물은 꿩의밥 줄기를 추출 대상으로 하여 얻어지는 모든 형태의 추출물로서, 예를 들어 식물 줄기에 직/간접적으로 열을 가해 얻는 진액 형태의 추출물, 살아있는 식물 줄기의 윗부분을 잘라 흘러나오는 진액 형태의 추출물, 식물을 유기용매, 또는 물로 추출하여 얻는 추출물, 식물 줄기를 연소시킬 때 기체를 냉각하여 얻는 추출물 등을 모두 포함하는 것이다.In the present invention, the pheasant rice stem extract refers to all types of extracts obtained by using pheasant rice stems as the extraction object, for example, extracts in the form of essence obtained by directly or indirectly heating plant stems, and living plant stems. It includes extracts in the form of a sap that flows out by cutting the upper part of the plant, extracts obtained by extracting plants with organic solvents or water, and extracts obtained by cooling the gas when burning plant stems.
본 발명에 있어서 상기 꿩의밥 열매 추출물은 유기 용매 또는 물을 사용하여 얻어지는 추출물과 꿩의밥 열매를 압착하여 얻어지는 진행 형태의 추출물, 용매 추출물을 감압하여 얻어지는 오일 추출물을 포함한다.In the present invention, the pheasant rice fruit extract includes an extract obtained using an organic solvent or water, a progressive extract obtained by pressing the pheasant rice fruit, and an oil extract obtained by reducing the solvent extract.
본 발명에 있어서 상기 꿩의밥 종자 추출물은 유기 용매 또는 물을 사용하여 얻어진 추출물과, 꿩의밥 종자를 압착하여 얻어지는 진액 추출물, 또는 용매 추출물을 감압하여 얻어진 오일 추출물을 포함한다.In the present invention, the pheasant rice seed extract includes an extract obtained using an organic solvent or water, a essence extract obtained by pressing pheasant rice seeds, or an oil extract obtained by reducing the solvent extract.
본 발명에 있어서, 상기 꿩의밥 잎 추출물은 유기 용매 또는 물 추출물을 의미한다. 예를 들어, 상기 꿩의밥 추출물은 각각의 줄기, 잎, 열매 및 종자를 분쇄한 후 유기 용매, 예를 들어, 50~100%의 탄소수 1 내지 10의 알코올, 에틸아세테이트, 헥산 및 디클로로메탄으로 이루어진 군으로부터 선택된 1종 이상의 추출용매로 추출된 추출물 또는 물 추출물일 수 있다.In the present invention, the pheasant rice leaf extract refers to an organic solvent or water extract. For example, the pheasant rice extract is obtained by pulverizing each stem, leaf, fruit, and seed, and then mixing it with an organic solvent, such as 50-100% alcohol with 1 to 10 carbon atoms, ethyl acetate, hexane, and dichloromethane. It may be an extract extracted with one or more extraction solvents selected from the group consisting of or a water extract.
추출물의 제조방법은 냉침 추출법, 온침 추출법 또는 열 추출법 등이 사용될 수 있으며, 통상의 추출기기, 초음파 분쇄 추출기 또는 분획기를 이용할 수 있다.The extract may be prepared using a cold needle extraction method, a warm needle extraction method, or a heat extraction method, and a conventional extraction device, an ultrasonic grinding extractor, or a fractionator can be used.
또 다른 예에 의하면, 상기 꿩의밥 추출물은 상기 열매 및/또는 종자의 유기용매 추출물을 감압하여 오일로 만든 것을 사용할 수도 있다.According to another example, the pheasant meal extract may be made into oil by depressurizing the organic solvent extract of the fruit and/or seed.
상기 제조된 추출물은 여과하거나 농축 또는 건조하여 용매를 제거할 수 있으며, 구체적으로 상기 여과는 여과지를 이용하거나 감압 여과기를 이용할 수 있으며, 상기 농축은 감압 농축기, 일예로 회전 증발기를 이용하여 감압 농축할 수 있고, 상기 건조는 일예로 동결 건조법으로 수행할 수 있다.The prepared extract may be filtered, concentrated, or dried to remove the solvent. Specifically, the filtration may be performed using filter paper or a reduced-pressure filter, and the concentration may be performed using a reduced-pressure concentrator, for example, a rotary evaporator. For example, the drying may be performed using a freeze-drying method.
본 발명에 있어서, 상기 추출물은 농축, 또는 희석하여 사용할 수 있고, 추출물의 증류액을 사용할 수도 있다. In the present invention, the extract can be used concentrated or diluted, and a distillate of the extract can also be used.
본 발명의 일 실시예에 있어서, 상기 피부외용제 조성물은 피부주름 방지용, 피부주름 개선용, 피부 톤 개선용 또는 피부모공 축소용 조성물일 수 있다.In one embodiment of the present invention, the composition for external application for skin may be a composition for preventing skin wrinkles, improving skin wrinkles, improving skin tone, or reducing skin pores.
상기 피부주름 방지, 피부주름 개선은 주름과 관련된 능력을 유지 또는 강화시키는 것을 의미하는 것으로, 콜라겐 당화반응을 감소시켜 효과적으로 최종당화산물의 생성을 억제하여 피부주름 등과 같은 피부노화 현상을 방지할 수 있으며, 상기 피부 톤 개선은 피부에서의 혈액의 순환을 돕고 피부 콜라겐 당화반응을 억제하여 투명하고, 균일한 피부 톤을 만들어 주는 것이다.Preventing skin wrinkles and improving skin wrinkles means maintaining or strengthening wrinkle-related abilities. By reducing the collagen glycation reaction, the production of advanced glycation end products can be effectively suppressed, thereby preventing skin aging phenomena such as skin wrinkles. , The skin tone improvement is to help blood circulation in the skin and inhibit skin collagen glycation reaction to create a transparent and uniform skin tone.
또한, 피부 모공이 넓어지는 원인은 피부에 탄력을 주고 지지대 역할을 하는 콜라겐과 엘라스틴의 힘이 떨어지게 되거나, 피부가 늘어져 모공이 커 보이는 것으로 피부 콜라겐 당화반응 억제 활성을 통해 피부 모공을 축소할 수 있다.In addition, the cause of enlarged skin pores is a decrease in the strength of collagen and elastin, which provide elasticity to the skin and act as a support, or the skin sags and the pores appear larger. Skin pores can be reduced by inhibiting the glycation reaction of skin collagen. .
또한, 본 발명에 따른 피부 외용제 조성물은 특히, 프로콜라겐 증가, 콜라겐 생합성 증가능을 가지고 있어, 피부 노화 방지, 피부 주름 개선/방지용으로 적용될 수 있으며, 이외에도 미백, 모공수축, 피지억제, 항균, iNOS 억제 활성을 통한 항염, 피부 염증 예방, 완화, 경감, 치료용으로 적용될 수 있다.In addition, the skin external composition according to the present invention has the ability to increase procollagen and collagen biosynthesis, and can be applied to prevent skin aging and improve/prevent skin wrinkles. In addition, it can be used for whitening, pore contraction, sebum inhibition, antibacterial, and iNOS. It can be applied for anti-inflammatory and skin inflammation prevention, relief, alleviation, and treatment through inhibitory activity.
본 발명의 일 실시예에 있어서, 상기 피부 외용제 조성물은 상기 꿩의밥 추출물 이외에 글리세린, 부틸렌글리콜, 프로필렌글키롤, 폴리옥시에틸렌 경화피마자유, 에탄올, 트리에탄올아민 등을 포함할 수 있으며, 방부제, 항료, 착색료, 정제수 등을 필요에 따라 미량 포함할 수 있다.In one embodiment of the present invention, the composition for external application for skin may include glycerin, butylene glycol, propylene glycol, polyoxyethylene hydrogenated castor oil, ethanol, triethanolamine, etc. in addition to the pheasant rice extract, a preservative, A small amount of flavoring, coloring, purified water, etc. may be included as needed.
본 발명에 있어서, 상기 피부 외용제 조성물은, 다양한 형태로 제조될 수 있는데, 예컨대, 화장수, 에센스, 젤, 에멀젼, 로션, 크림(수중유적형, 유중수적형, 다중상), 용액, 현탁액(무수 및 수계), 무수 생성물(오일 및 글리콜계), 젤, 마스크, 팩, 분말, 또는 젤라틴 등의 피막이 있는 캅셀(소프트 캅셀, 하드 캅셀), 연고, 패취, 파운데이션, 메이크업베이스, 자외선차단제, 입욕제, 비누, 액체 세정료, 분무제 제형 등의 형태로 제조될 수 있다.In the present invention, the composition for external skin application can be prepared in various forms, such as lotion, essence, gel, emulsion, lotion, cream (oil-in-water type, water-in-oil type, multi-phase), solution, suspension (anhydrous) and water-based), anhydrous products (oil and glycol-based), gels, masks, packs, powders, capsules with a coating such as gelatin (soft capsules, hard capsules), ointments, patches, foundations, makeup bases, sunscreens, bath additives, It can be manufactured in the form of soap, liquid detergent, spray formulation, etc.
본 발명에 있어서 피부는 얼굴뿐만 아니라, 두피, 전신도 포함되는 개념으로, 이러한 두피에 적용될 수 있는 피부 외용제 조성물로써, 샴푸, 린스, 트리트먼트, 발모제 등이 있고, 전신에 적용될 수 있는 바디클렌져 등의 용도로써 다양한 형태로 제조될 수 있다.In the present invention, skin includes not only the face, but also the scalp and the entire body. External skin compositions that can be applied to the scalp include shampoos, rinses, treatments, hair growth agents, etc., and body cleansers that can be applied to the entire body. It can be manufactured in various forms for various purposes.
본 발명에 있어서, 상기 꿩의밥 추출물을 함유 피부 외용제 조성물의 제조방법은 전술한 제조방법에 한정되는 것은 아니며, 본 발명이 속하는 기술분야에서 통상의 지식을 가진 자라면 상기 제조방법을 일부 변형시킨 방법으로도 본 발명에 따른 꿩의밥 추출물을 함유 피부 외용제 조성물을 제조할 수 있다.In the present invention, the manufacturing method of the skin external composition containing the pheasant rice extract is not limited to the above-mentioned manufacturing method, and those skilled in the art will know that the manufacturing method has been partially modified. A composition for external use for skin containing pheasant rice extract according to the present invention can also be prepared by this method.
특히, 상기 피부 외용제 조성물은 본 발명에 특별히 개시된 제조방법 이외에도, 통상적으로 알려진 제조방법을 이용하여, 일반적인 유화 제형 및 가용화 제형의 형태로 제조될 수 있다. 화장료 조성물로 제조될 경우, 유화 제형의 화장품으로는 영양화장수, 크림, 에센스 등이 있으며, 가용화 제형의 화장품으로는 유연화장수가 있다. 또한, 피부과학적으로 허용 가능한 매질 또는 기제를 함유함으로써 피부과 학 분야에서 통상적으로 사용되는 국소적용 또는 전신적용할 수 있는 보조제 형태로 제조될 수 있다. 또한, 적합한 화장품의 제형으로는, 예를 들면 용액, 겔, 고체 또는 반죽 무수 생성물, 수상에 유상을 분산시켜 얻은 에멀젼, 현탁액, 마이크로에멀젼, 마이크로캡슐, 미세과립구 또는 이온형(리포좀), 비이온형의 소낭 분산제의 형태, 크림, 스킨, 로션, 파우더, 연고, 스프레이 또는 콘실스틱의 형태로 제공될 수 있다. 또한, 포말(foam)의 형태 또는 압축된 추진제를 더 함유한 에어로졸 조성물의 형태로도 제조될 수 있다. 또한, 본 발명의 피부 외용제 조성물은 추가로 지방 물질, 유기 용매, 용해제, 농축제 및 겔화제, 연화제, 항산화제, 현탁화제, 안정화제, 발포제(foaming agent), 방향제, 계면활성제, 물, 이온형 또는 비이온형 유화제, 충전제, 금속이온봉쇄제 및 킬레이트화제, 보존제, 비타민, 차단제, 습윤화제, 필수 오일, 염료, 안료, 친수성 또는 친유성 활성제, 지질 소낭 또는 화장품에 통상적으로 사용되는 임의의 다른 성분과 같은 화장품학 또는 피부과학 분야에서 통상적으로 사용되는 보조제를 함유할 수 있다. 그리고 상기의 성분들은 피부과학 분야에서 일반적으로 사용되는 양으로 도입될 수 있다. In particular, the composition for topical skin application can be prepared in the form of a general emulsified formulation and solubilized formulation using commonly known production methods in addition to the production method specifically disclosed in the present invention. When manufactured as a cosmetic composition, emulsified cosmetics include nutritional lotion, cream, essence, etc., and solubilized cosmetics include soft lotion. In addition, by containing a dermatologically acceptable medium or base, it can be manufactured in the form of an adjuvant that can be applied topically or systemically, which is commonly used in the field of dermatology. In addition, suitable cosmetic formulations include, for example, solutions, gels, solid or pasty anhydrous products, emulsions obtained by dispersing the oil phase in the water phase, suspensions, microemulsions, microcapsules, microgranules or ionic (liposomes), non-ionic. It may be provided in the form of a vesicular dispersion, cream, skin, lotion, powder, ointment, spray or conceal stick. Additionally, it can be prepared in the form of foam or in the form of an aerosol composition further containing compressed propellant. In addition, the composition for topical skin application of the present invention further contains fatty substances, organic solvents, solubilizers, thickening and gelling agents, softeners, antioxidants, suspending agents, stabilizers, foaming agents, fragrances, surfactants, water, and ions. Formal or non-ionic emulsifiers, fillers, sequestering and chelating agents, preservatives, vitamins, blocking agents, wetting agents, essential oils, dyes, pigments, hydrophilic or lipophilic active agents, lipid vesicles or any of those commonly used in cosmetics. It may contain other ingredients and auxiliaries commonly used in the fields of cosmetology or dermatology. And the above ingredients can be introduced in amounts commonly used in the field of dermatology.
본 발명에 있어서, 상기 피부 외용제 조성물은 화장료 조성물 또는 약제학적 조성물일 수 있다.In the present invention, the external skin composition may be a cosmetic composition or a pharmaceutical composition.
상기 화장료 조성물에 있어서는, 화장품 제제에 있어서 수용 가능한 담체를 포함할 수 있다. 여기서, "화장품 제제에 있어서 수용 가능한 담체"란 화장품 제제에 포함될 수 있는 이미 공지되어 사용되고 있는 화합물 또는 조성물이거나 앞으로 개발될 화합물 또는 조성물로서 피부와의 접촉시 인체가 적응 가능한 이상의 독성, 불안정성 또는 자극성이 없는 것을 말한다.The cosmetic composition may contain a carrier acceptable in cosmetic preparations. Here, “acceptable carrier in cosmetic preparations” means a compound or composition that is already known and used or a compound or composition that will be developed in the future that can be included in cosmetic preparations and has no toxicity, instability or irritation beyond what the human body can adapt to upon contact with the skin. It says something that doesn't exist.
상기 담체는 본 발명의 피부 외용제 조성물에 그것의 전체 중량에 대하여 약 1중량% 내지 약 99.99중량%, 바람직하게는 조성물의 중량의 약 90중량% 내지 약 99.99중량%로 포함될 수 있다. 그러나 상기 비율은 본 발명의 피부 외용제 조성물이 제조되는 후술한 바의 제형에 따라 또 그것의 구체적인 적용 부위(얼굴, 목 등)나 그것의 바람직한 적용량 등에 따라 달라지는 것이기 때문에, 상기 비율은 어떠한 측면으로든 본 발명의 범위를 제한하는 것으로 이해되어서는 안 된다.The carrier may be included in the external skin preparation composition of the present invention in an amount of about 1% to about 99.99% by weight based on the total weight, and preferably in an amount of about 90% by weight to about 99.99% by weight of the composition. However, since the above ratio varies depending on the formulation as described below in which the external skin composition of the present invention is manufactured, its specific application site (face, neck, etc.), and its preferred application amount, the above ratio can be viewed in any way. It should not be construed as limiting the scope of the invention.
상기 담체로서는 알코올, 오일, 계면활성제, 지방산, 실리콘 오일, 습윤제, 보습제, 점성 변형제, 유제, 안정제, 자외선 산란제, 자외선흡수제, 발색제, 향료 등이 예시될 수 있다. 상기 알코올, 오일, 계면활성제, 지방산, 실리콘 오일, 습윤제, 보습제, 점성 변형제, 유제, 안정제, 자외선산란제, 자외선흡수제, 발색제, 향료로 사용될 수 있는 화합물/조성물 등은 이미 당업계에 공지되어 있기 때문에 당업자라면 적절한 해당 물질/조성물을 선택하여 사용할 수 있다.Examples of the carrier include alcohol, oil, surfactant, fatty acid, silicone oil, wetting agent, moisturizing agent, viscosity modifier, emulsion, stabilizer, ultraviolet light scattering agent, ultraviolet absorber, coloring agent, fragrance, etc. Compounds/compositions that can be used as alcohols, oils, surfactants, fatty acids, silicone oils, wetting agents, moisturizers, viscosity modifiers, emulsions, stabilizers, UV scattering agents, UV absorbers, coloring agents, and fragrances are already known in the art. Therefore, a person skilled in the art can select and use the appropriate material/composition.
본 발명에 있어서, 상기 약제학적 조성물은 피부질환 치료, 상처치유 및 피부재생용 조성물로써, 로션, 연고, 겔, 크림, 패취 또는 분무제와 같은 경피 투여형 제형일 수 있으나, 이에 제한되지 않는다.In the present invention, the pharmaceutical composition is a composition for treating skin diseases, wound healing, and skin regeneration, and may be a transdermal dosage form such as lotion, ointment, gel, cream, patch, or spray, but is not limited thereto.
이하, 하기 구체예들을 통하여 본 발명을 더욱 상세하게 설명하지만, 이는 본 발명의 예시일 뿐 본 발명의 권리범위가 이에 한정되는 것은 아니다.Hereinafter, the present invention will be described in more detail through the following specific examples, but these are only examples of the present invention and the scope of the present invention is not limited thereto.
[[ 제조예Manufacturing example ] ] 꿩의밥Pheasant's Rice 추출물 제조 Extract Preparation
꿩의밥 전초 100 g을 수거하여 70% 에탄올 수용액 5 L를 넣고 실온에서 48시간 동안 냉침한 후 여과 및 감압 농축하여 추출물 15 g을 얻었다. 세포 실험을 진행하기 위해 추출물을 DMSO(Dimethyl sulfoxide)에 녹여 시료를 제조하였다.100 g of pheasant rice starter was collected, 5 L of 70% ethanol aqueous solution was added, cold soaked at room temperature for 48 hours, filtered, and concentrated under reduced pressure to obtain 15 g of extract. To conduct cell experiments, samples were prepared by dissolving the extract in DMSO (Dimethyl sulfoxide).
[[ 시험예Test example 1] One] 꿩의밥Pheasant's Rice 추출물의 항산화력 측정 Measurement of antioxidant power of extract
[[ 시험예Test example 1-1] 총 폴리페놀( 1-1] Total polyphenols ( polyphenolpolyphenols )함량 측정)Content measurement
폴리페놀은 몸에 있는 활성산소를 해가 없는 물질로 바꿔주는 항산화물질 중 하나이다. 본 발명의 꿩의밥 추출물의 총 폴리페놀 함량을 측정하기 위하여 폴린-데니스(Folin-Denis) 법을 일부 변형하여 폴린-시약(Folin-reagent)이 추출물의 페놀성 화합물에 의해 환원되면 몰리브덴의 청색으로 발색되는 원리를 이용하여 정량하였다. 꿩의밥 추출물(1 mg/ml) 100 ㎕에 폴린-시오칼토 시약(folin-ciocalteu reagent) 200 ㎕을 가한 후 700 mM Na2CO3용액 800 ㎕를 혼합하고 실온에서 40분 방치한 후 microplate reader를 이용하여 750 nm에서 흡광도를 측정하였다. 갈릭 산(Gallic acid, sigma)를 표준물질로 사용하여 0~500 μg/ml 농도로 제조한 후 시료와 동일한 방법으로 분석하여 얻은 표준검량 선으로부터 꿩의밥 추출물의 폴리페놀 함량을 분석하고, 그 결과를 갈릭산 당량 (GAE, gallic acid equivalents)으로 표 1에 나타내었다.Polyphenol is one of the antioxidant substances that changes free radicals in the body into harmless substances. In order to measure the total polyphenol content of the pheasant rice extract of the present invention, the Folin-Denis method is partially modified to obtain a blue color of molybdenum when the Folin-reagent is reduced by the phenolic compounds of the extract. It was quantified using the principle of color development. Add 200 μl of folin-ciocalteu reagent to 100 μl of pheasant rice extract (1 mg/ml), mix with 800 μl of 700 mM Na 2 CO 3 solution, leave at room temperature for 40 minutes, and then use a microplate reader. Absorbance was measured at 750 nm using . Gallic acid (sigma) was used as a standard material to prepare the concentration of 0-500 μg/ml, and the polyphenol content of the pheasant rice extract was analyzed from the standard calibration line obtained by analyzing in the same manner as the sample. The results are shown in Table 1 in terms of gallic acid equivalents (GAE).
(μg GAE/mg 꿩의밥 추출물)Total polyphenol content
(μg GAE/mg pheasant meal extract)
표 1에서 나타나는 바와 같이, 1 mg의 꿩의밥 추출물은 폴리페놀을 총 63.63 μg을 함유하고 있어 꿩의밥 추출물의 총 폴리페놀 함량이 높음을 알 수 있다.As shown in Table 1, 1 mg of pheasant rice extract contains a total of 63.63 μg of polyphenols, indicating that the total polyphenol content of the pheasant rice extract is high.
[[ 시험예Test example 1-2] 1-2] DPPHDPPH 항산화 효과 측정 Measurement of antioxidant effect
DPPH(1,1-diphenyl-2-picrydrazyl) 법에 의한 항산화 활성을 측정하였다. DPPH법은 DPPH라는 유리기를 사용하여 환원력에 의한 자유라디칼 소거활성을 측정하는 것이다.Antioxidant activity was measured by the DPPH (1,1-diphenyl-2-picrydrazyl) method. The DPPH method measures free radical scavenging activity by reducing power using a free radical called DPPH.
96-웰 플레이트(well-plate)에 연속희석법(serial dilution)으로 각 시료 50 ㎕를 넣은 후, 0.2 mM DPPH(Sigma, St. Louis, USA) 용액 100 ㎕를 첨가하여 30분간 반응을 시켰다. 시료에 의해 DPPH가 환원되어 흡광도가 감소하는 정도를 대조군의 흡광도와 비교하여 파장 517 nm에서 자유라디칼 소거율을 측정하였고, 그 결과를 표 1에 나타내었다.After adding 50 ㎕ of each sample to a 96-well plate by serial dilution, 100 ㎕ of 0.2 mM DPPH (Sigma, St. Louis, USA) solution was added and allowed to react for 30 minutes. The degree to which DPPH was reduced by the sample and the absorbance decreased was compared with the absorbance of the control group, and the free radical scavenging rate was measured at a wavelength of 517 nm, and the results are shown in Table 1.
자유라디칼 소거활성(scavenging activity, %)은 다음의 수학식 1로 산출하였다.Free radical scavenging activity (%) was calculated using Equation 1 below.
<수학식 1><Equation 1>
자유라디칼 소거활성 (%) = (A - B) / A X 100Free radical scavenging activity (%) = (A - B) / A
A : 시료를 처리하지 않은 대조군의 흡광도A: Absorbance of control group without sample treatment
B : 시료를 처리한 시험군의 흡광도B: Absorbance of the test group treated with the sample
자유라디칼 소거활성이 50% 감소하는데 필요한 시료의 농도를 SC 50으로 나타냈다.The sample concentration required to reduce free radical scavenging activity by 50% was expressed as SC 50.
상기 표 2에서 볼 수 있는 바와 같이, 꿩의밥 추출물은 항산화 실험에서 대조군으로 사용된 대표적인 항산화 물질인 라스베라트롤의 약 20% 수준의 우수한 항산화 효과를 나타내었다.As can be seen in Table 2, the pheasant rice extract showed an excellent antioxidant effect of about 20% of that of resveratrol, a representative antioxidant used as a control in the antioxidant experiment.
[[ 시험예Test example 2] 단백질 2] Protein 당화Glycosylation (protein (protein glycationglycation ) 반응 저해 효과 측정) Measurement of response inhibition effect
우혈청 알부민(bovine serum albumin, BSA; Sigma, St. Louis, USA)을 반응 기질로 하여 당화 반응을 수행하고 형광 변화 정도를 측정함으로써 각 시료의 단백질 당화 반응 저해 효과를 측정하였다.The glycation reaction was performed using bovine serum albumin (BSA; Sigma, St. Louis, USA) as a reaction substrate, and the degree of fluorescence change was measured to determine the effect of inhibiting the protein glycation reaction of each sample.
우선 글리옥살(glyoxal) 용액(100 mM) 0.4 ml과 PBS buffer (pH 7.4)에 희석한 500 ppm 농도의 각 시료 용액 0.4 ml을 첨가하여 37℃에서 2시간동안 반응시킨 후, 0.2 ml BSA 용액 (20 mg/ml)을 넣어 추가로 48시간동안 같은 온도에서 반응시켰다. 또한 0.02% 소디움 아자이드(sodium azide)를 넣어 반응기간 동안 미생물의 오염을 방지하였다. 각 시료에 의한 형광도의 간섭을 배제하기 위하여 BSA를 처리하지 않은 시료 처리군도 함께 제조하여 같은 기간 반응시켰다. 시료의 경우 당화반응이 일어나지 않은 각 시료의 형광도를 배제한 값을 사용하였다. 양성대조군으로는 아미노구아니딘 염산염(aminoguanidine-HCl)을 사용하였고, 반응 후 Victor X2 형광분광분석기(Perkin Elmer Inc.)를 이용하여 들뜸파장(Ex: Excitation wavelength) 355 nm, 방출파장(Em: Emission wavelength) 485 nm로 조정하여 형광도를 측정하였다.First, 0.4 ml of glyoxal solution (100 mM) and 0.4 ml of each sample solution with a concentration of 500 ppm diluted in PBS buffer (pH 7.4) were added and reacted at 37°C for 2 hours, followed by 0.2 ml of BSA solution ( 20 mg/ml) was added and reacted at the same temperature for an additional 48 hours. Additionally, 0.02% sodium azide was added to prevent microbial contamination during the reaction period. To exclude interference in fluorescence by each sample, a sample treatment group not treated with BSA was also prepared and reacted for the same period. In the case of samples, the value excluding the fluorescence of each sample in which glycation reaction did not occur was used. Aminoguanidine-HCl was used as a positive control, and after reaction, the excitation wavelength (Ex: Excitation wavelength) was 355 nm and the emission wavelength (Em: Emission wavelength) was measured using a Victor ) Fluorescence was measured by adjusting to 485 nm.
최종당화산물 생성 억제율은 다음의 수학식 2로 산출하였다.The inhibition rate of advanced glycation end products was calculated using Equation 2 below.
<수학식 2><Equation 2>
최종당화산물 생성 억제율 (%) = 100- {(샘플처리군-블랭크값)/(음성대조군-블랭크값)}X100 Advanced glycation end product production inhibition rate (%) = 100- {(sample treatment group-blank value)/(negative control group-blank value)}X100
(Aminoguanidine HCl)
(양성대조군)Aminoguanidine Hydrochloride
(Aminoguanidine HCl)
(Positive control group)
상기 표 3에서 알 수 있는 바와 같이, 500 ppm의 꿩의밥 추출물에서 17.1%의 최종당화산물 생성 저해율이 확인되었으며, 같은 농도의 아미노구아니딘 염산염의 약 26.5% 수준의 우수한 당화반응 억제효과를 나타내었다.As can be seen in Table 3, an inhibition rate of advanced glycation end products of 17.1% was confirmed in 500 ppm of pheasant rice extract, and it showed an excellent glycation reaction inhibition effect of about 26.5% of aminoguanidine hydrochloride at the same concentration. .
[[ 시험예Test example 3] 3] 꿩의밥Pheasant's Rice 추출물의 피부 섬유아세포 생존율 측정 Measurement of skin fibroblast survival rate of extract
본 발명의 꿩의밥 추출물의 인간 정상 피부 섬유아세포(CCD-986sk)에 대한 세포독성을 측정하기 위해 세포를 1X104 cells/well 농도로 96-well plate에 분주한 후 37℃, 5% CO2 조건에서 24시간 동안 배양하였다. 배양배지를 제거 후 무혈청배지에 희석한 10~100 ppm 농도의 꿩의밥 추출물을 세포에 처리하여 24시간 동안 추가 배양하였다. 배양 후, 세포 생존율을 확인하기 위하여 배양배지를 제거 후 각 well 당 0.5 mg/ml 농도의 MTT용액 200 ㎕를 처리하여 3시간동안 반응시켰다. 생성된 포마잔(formazan)을 DMSO 100 ㎕로 용해시켜 micro-plate reader를 이용하여 540 nm 파장에서 흡광도를 측정하였다. 정상 대조군으로는 추출물을 처리하지 않은 well의 세포 생존율을 100%으로 하여 꿩의밥 추출물 처리군의 세포 생존율을 수학식 3에 따라 계산하여 산출한 결과를 도 1에 나타내었다.To measure the cytotoxicity of the pheasant meal extract of the present invention on human normal skin fibroblasts (CCD-986sk), the cells were distributed in a 96-well plate at a concentration of 1X10 4 cells/well and incubated at 37°C, 5% CO 2 It was cultured for 24 hours under these conditions. After removing the culture medium, the cells were treated with pheasant rice extract at a concentration of 10 to 100 ppm diluted in serum-free medium and cultured for an additional 24 hours. After culturing, to check cell viability, the culture medium was removed, and each well was treated with 200 ㎕ of MTT solution at a concentration of 0.5 mg/ml and reacted for 3 hours. The produced formazan was dissolved in 100 μl of DMSO, and the absorbance was measured at a wavelength of 540 nm using a micro-plate reader. As a normal control group, the cell survival rate of wells not treated with the extract was set as 100%, and the cell survival rate of the pheasant rice extract-treated group was calculated according to Equation 3, and the calculated results are shown in Figure 1.
<수학식 3><Equation 3>
섬유아세포 생존율(%) = (A/B) X 100Fibroblast survival rate (%) = (A/B)
A: 시료를 처리한 시험군의 흡광도A: Absorbance of the test group treated with the sample
B: 시료를 처리하지 않은 대조군의 흡광도B: Absorbance of control group without sample treatment
도 1에 나타낸 바와 같이, 꿩의밥 추출물을 10~100 ppm 피부 섬유아세포에 처리하였을 때 100 ppm까지 독성이 보이지 않은 것으로 확인되었다.As shown in Figure 1, when 10 to 100 ppm of pheasant rice extract was treated with skin fibroblasts, it was confirmed that no toxicity was observed up to 100 ppm.
[[ 시험예Test example 4] 4] 당화스트레스에In glycation stress 의한 콜라겐 by collagen 당화Glycosylation 저해 효과 측정 Measurement of inhibition effect
시험예 3과 같은 방법으로 세포 분주를 하여 24시간 배양을 한 뒤, 배양배지를 제거 후 무혈청배지에 희석한 10, 25, 50 ppm 농도의 꿩의밥 추출물을 세포에 처리하여 24시간 동안 추가 배양하였다. 24시간 후, 무혈청배지에 희석한 0.5 mM의 글리옥살(glyoxal)을 48시간 처리하여 세포에 당화스트레스를 유발하였다. 48시간 반응이 끝난 후, 각 웰의 상층액을 모아 Human pro-collagen 1 alpha 1 분석 키트 (R&D system, DY6220-05)를 이용하여 상층액의 프로콜라겐 함량(pg/ml)을 측정하였다. 꿩의밥 추출물을 처리시 콜라겐 당화 억제로 인한 콜라겐 회복률(%)을 구하기 위해 정상 대조군으로는 시료와 글리옥살을 처리하지 않은 well의 상층액 내 콜라겐 함량을 100%으로 하여 정상 대조군 대비 글리옥살만 처리군 (음성대조군)과 시료 및 글리옥살 처리군의 콜라겐 생성률(%)을 계산하여 결과를 표 4에 나타내었다.Cells were distributed in the same manner as in Test Example 3 and cultured for 24 hours. After removing the culture medium, pheasant rice extract at concentrations of 10, 25, and 50 ppm diluted in serum-free medium was treated with the cells and added for 24 hours. Cultured. After 24 hours, cells were treated with 0.5 mM glyoxal diluted in serum-free medium for 48 hours to induce glycation stress. After the 48-hour reaction was completed, the supernatant from each well was collected and the procollagen content (pg/ml) of the supernatant was measured using the Human pro-collagen 1 alpha 1 analysis kit (R&D system, DY6220-05). To determine the collagen recovery rate (%) due to inhibition of collagen glycation when treated with pheasant rice extract, the collagen content in the supernatant of the sample and the well not treated with glyoxal was set as 100% as the normal control, and only glyoxal was compared to the normal control. The collagen production rate (%) of the treatment group (negative control group), sample, and glyoxal treatment group was calculated, and the results are shown in Table 4.
콜라겐 생성률(%)Compared to normal control group
Collagen production rate (%)
콜라겐 생성 회복률(%)Compared to negative control group
Collagen production recovery rate (%)
(glyoxal)
0.5 mMglyoxal
(glyoxal)
0.5mM
추출물Pheasant's Rice
extract
표 4에서 알 수 있는 바와 같이, 음성대조군에서 정상대조군 대비 71.61%의 콜라겐 생성률을 보여 세포에 당화스트레스를 유도시 콜라겐 생합성 저해 및 콜라겐 당화가 일어나는 것을 확인하였다. 한편 꿩의밥 추출물을 10, 25, 50ppm 농도로 선처리 후 당화스트레스를 유도시 농도의존적으로 음성대조군 대비 콜라겐 생성률이 증가하였으며, 이에 따라 콜라겐 생성 회복률 또한 유의적으로 증가하는 것을 확인하였다. 상기 결과에 따라, 꿩의밥 추출물은 우수한 피부 콜라겐 당화 억제 활성을 나타냄을 확인하였다. As can be seen in Table 4, the negative control group showed a collagen production rate of 71.61% compared to the normal control group, confirming that collagen biosynthesis is inhibited and collagen glycation occurs when glycation stress is induced in cells. Meanwhile, when glycation stress was induced after pretreatment with pheasant rice extract at concentrations of 10, 25, and 50 ppm, the collagen production rate increased compared to the negative control group in a concentration-dependent manner, and the collagen production recovery rate was also confirmed to significantly increase accordingly. According to the above results, it was confirmed that the pheasant rice extract exhibits excellent skin collagen glycation inhibition activity.
또한, 표 4에는 명시하지 않았지만 0.5mM의 글리옥살(glyoxal)을 48시간 처리시 정상대조군 대비 90% 이상의 세포 생존율을 확인하여 세포 사멸에 크게 영향을 미치지 않은 글리옥살 농도 내에서 세포에 당화스트레스를 유발하였다. In addition, although not specified in Table 4, when treated with 0.5mM glyoxal for 48 hours, a cell survival rate of more than 90% compared to the normal control was confirmed, showing that glycation stress was applied to cells within the glyoxal concentration that did not significantly affect cell death. It was caused.
[[ 시험예Test example 5] 5] 당화스트레스에In glycation stress 의한 섬유아세포 내 within fibroblasts by carboxymethylcarboxymethyl lysine( lysine( CMLCML ) 생성 저해효과 측정) Measurement of production inhibition effect
본 발명의 꿩의밥 추출물의 피부 섬유아세포 내 최종당화산물인 CML의 생성 저해효과를 측정하기 위해 세포를 4x105 cells/well 농도로 100 mm dish에 분주한 후 37℃, 5% CO2 조건에서 24시간 동안 배양하였다. 배양배지를 제거 후 무혈청배지에 희석한 25, 50, 100 ppm 농도의 꿩의밥 추출물과 10 ppm 농도의 아미노구아니딘 염산염을 세포에 처리하여 24시간 추가 배양하였다. 24시간 후, 무혈청배지에 희석한 0.5 mM의 글리옥살(glyoxal)을 72시간 처리하여 세포에 당화스트레스를 유발하였다. 72시간 반응이 끝난 세포는 PBS buffer로 2회 세척한 후 RIPA lysis buffer를 첨가하고 4℃에서 5분간 lysis 시킨 후 12,000 rpm에서 10분간 원심분리하였다. 상층액을 취하여 동일하게 단백질 정량한 후 CML competitive ELISA kit (Cell biolabs, STA-816)를 이용하여 상층액의 CML 함량(μg/ml)을 측정하였다. 꿩의밥 추출물 처리시 피부 섬유아세포 당화 억제로 인한 세포 내 CML 생성 억제 정도를 확인하기 위해 글리옥살만 처리한 음성 대조군의 CML 함량을 100%으로 하여, 음성 대조군 대비 정상 대조군(시료와 글리옥살 무첨가)과 꿩의밥 추출물 및 아미노구아니딘 염산염 처리군의 CML 생성률(%)을 계산하여 결과를 도 2에 나타내었다.To measure the inhibitory effect of the pheasant rice extract of the present invention on the production of CML, an advanced glycation product, in skin fibroblasts, cells were seeded in a 100 mm dish at a concentration of 4x10 5 cells/well and incubated at 37°C and 5% CO 2 conditions. Cultured for 24 hours. After removing the culture medium, the cells were treated with pheasant rice extract at concentrations of 25, 50, and 100 ppm and aminoguanidine hydrochloride at a concentration of 10 ppm diluted in serum-free medium, and cultured for an additional 24 hours. 24 hours later, cells were treated with 0.5 mM glyoxal diluted in serum-free medium for 72 hours to induce glycation stress. After 72 hours of reaction, the cells were washed twice with PBS buffer, RIPA lysis buffer was added, lysed for 5 minutes at 4°C, and then centrifuged at 12,000 rpm for 10 minutes. The supernatant was taken and the protein was quantified in the same manner, and then the CML content (μg/ml) of the supernatant was measured using a CML competitive ELISA kit (Cell biolabs, STA-816). In order to confirm the degree of inhibition of intracellular CML production due to inhibition of skin fibroblast glycation when treated with pheasant rice extract, the CML content of the negative control group treated with glyoxal only was set at 100%, and the normal control group (sample and no glyoxal added) was compared to the negative control group. ) and the CML production rate (%) of the pheasant rice extract and aminoguanidine hydrochloride treated groups were calculated and the results are shown in Figure 2.
도 2에 나타낸 바와 같이, 피부 섬유아세포에 0.5 mM의 글리옥살을 처리시 정상대조군 대비 4배가량 CML 생성률이 증가되었으며, 꿩의밥 추출물을 25, 50, 100 ppm 처리하였을 때 음성 대조군 대비 CML 생성률이 각각 17.6%, 18.31%, 21.18%씩 억제되는 것을 확인하였다. 아미노구아니딘 염산염은 농도 별로 24시간 처리하였을 때 10 ppm까지 세포 독성을 보이지 않아 해당 농도를 세포에 처리하였으며, CML 생성률이 21.66% 억제되는 것을 확인하였다. 꿩의밥 추출물의 세포 독성이 나타나지 않은 농도 범위 한에서는 100 ppm에서 최고 21.18%으로 10 ppm의 아미노구아니딘 염산염과 비슷한 수준의 CML 생성 억제 정도를 보임으로써 훨씬 안전하면서 피부 세포 당화 억제 효과가 우수한 것을 확인하였다.As shown in Figure 2, when skin fibroblasts were treated with 0.5 mM glyoxal, the CML production rate increased by about 4 times compared to the normal control, and when pheasant rice extract was treated at 25, 50, and 100 ppm, the CML production rate was increased compared to the negative control group. It was confirmed that this was suppressed by 17.6%, 18.31%, and 21.18%, respectively. Aminoguanidine hydrochloride did not show cytotoxicity up to 10 ppm when treated for 24 hours at each concentration, so cells were treated at that concentration, and it was confirmed that the CML production rate was inhibited by 21.66%. Within the concentration range where pheasant rice extract did not show cytotoxicity, it showed a similar level of inhibition of CML formation as 10 ppm aminoguanidine hydrochloride, up to 21.18% at 100 ppm, confirming that it was much safer and had an excellent effect in inhibiting skin cell glycation. .
상기 실험 결과에 따르면, 본 발명에 따른 꿩의밥 추출물을 처리시 항산화 활성, 콜라겐 당화 억제, 그리고 최종당화산물 생성 억제하는 효과가 있음을 확인하였다. 특히 단백질 당화 억제제로 알려져 있는 아미노구아니딘 염산염보다 세포 독성은 적으면서, 세포내 당화 억제효과가 비슷한 것을 확인하여 본 발명에 따른 조성물은 피부 내 당화스트레스로 유도된 피부노화, 주름 및 피부 투명감 저하를 효과적으로 개선시킬 수 있다.According to the above experimental results, it was confirmed that the treatment of the pheasant rice extract according to the present invention had the effect of inhibiting antioxidant activity, collagen glycation, and production of advanced glycation products. In particular, it was confirmed that the composition according to the present invention has less cytotoxicity than aminoguanidine hydrochloride, which is known as a protein glycation inhibitor, and has a similar inhibitory effect on intracellular glycation, effectively preventing skin aging, wrinkles, and decreased skin transparency induced by glycation stress in the skin. It can be improved.
이하, 상기한 실험예의 결과를 근거로 하여, 꿩의밥 추출물 추출물을 이용한 여러 제형예를 조성하여 제시한다. 그러나 이들 제형예는 본 발명을 설명하기 위한 것으로, 본 발명의 제형이 이들 제형예에만 국한되지 않는다는 것은 당업계에서 통상의 지식을 가진자에게 있어서 자명한 것이다.Below, based on the results of the above-described experimental examples, several formulation examples using pheasant rice extract extract are prepared and presented. However, these formulation examples are for illustrating the present invention, and it is obvious to those skilled in the art that the formulation of the present invention is not limited to these formulation examples.
Claims (10)
A cosmetic composition for preventing skin wrinkles, improving skin wrinkles, or reducing skin pores, comprising pheasant rice extract as an active ingredient.
상기 꿩의밥 추출물은 상기 꿩의밥의 잎, 꽃, 열매, 줄기, 뿌리 및 종자로 이루어진 군으로부터 선택된 1종 이상의 추출물인 것을 특징으로 하는 피부주름 방지용, 피부주름 개선용, 또는 피부모공 축소용 화장료 조성물.
According to paragraph 1,
The pheasant rice extract is for preventing skin wrinkles, improving skin wrinkles, or reducing skin pores, characterized in that it is one or more extracts selected from the group consisting of leaves, flowers, fruits, stems, roots, and seeds of the pheasant rice. Cosmetic composition.
상기 꿩의밥 추출물은 피부 콜라겐 당화반응(collagen glycation) 및 최종당화산물(advanced glycation end products, AGEs)의 생성을 억제시키는 것을 특징으로 하는 피부주름 방지용, 피부주름 개선용, 또는 피부모공 축소용 화장료 조성물.
According to paragraph 1,
The pheasant rice extract is a cosmetic for preventing skin wrinkles, improving skin wrinkles, or reducing skin pores, which is characterized by inhibiting skin collagen glycation and the production of advanced glycation end products (AGEs). Composition.
상기 꿩의밥 추출물은 상기 화장료 조성물 전체 중량에 대하여 0.0001 내지 30중량%로 포함되는 것을 특징으로 하는 피부주름 방지용, 피부주름 개선용, 또는 피부모공 축소용 화장료 조성물.
According to paragraph 1,
A cosmetic composition for preventing skin wrinkles, improving skin wrinkles, or reducing skin pores, characterized in that the pheasant rice extract is contained in an amount of 0.0001 to 30% by weight based on the total weight of the cosmetic composition.
상기 꿩의밥 추출물은 물, 유기용매 또는 이들의 혼합물로 추출되는 것을 특징으로 하는 피부주름 방지용, 피부주름 개선용, 또는 피부모공 축소용 화장료 조성물.
According to paragraph 1,
A cosmetic composition for preventing skin wrinkles, improving skin wrinkles, or reducing skin pores, characterized in that the pheasant rice extract is extracted with water, an organic solvent, or a mixture thereof.
상기 유기용매는 탄소수 1 내지 10의 무수 또는 함유 메탄올, 에탄올, 프로필알코올, 부틸알코올, 글리세린, 프로필렌글리콜 및 부틸렌글리콜로 이루어진 군으로부터 선택된 1종 이상인 것을 특징으로 하는 피부주름 방지용, 피부주름 개선용, 또는 피부모공 축소용 화장료 조성물.
According to clause 5,
The organic solvent is used for preventing skin wrinkles and improving skin wrinkles, wherein the organic solvent is at least one selected from the group consisting of anhydrous or containing carbon atoms of 1 to 10 methanol, ethanol, propyl alcohol, butyl alcohol, glycerin, propylene glycol, and butylene glycol. , or a cosmetic composition for reducing skin pores.
상기 화장료 조성물은 화장수, 에센스, 로션, 크림, 팩, 젤, 연고, 패취, 파운데이션, 메이크업베이스, 자외선차단제, 입욕제, 비누, 액체 세정료 및 분무제로 이루어진 군으로부터 선택된 1종 이상의 제형인 것을 특징으로 하는 피부주름 방지용, 피부주름 개선용, 또는 피부모공 축소용 화장료 조성물.
According to paragraph 1,
The cosmetic composition is characterized in that it is one or more formulations selected from the group consisting of lotion, essence, lotion, cream, pack, gel, ointment, patch, foundation, makeup base, sunscreen, bath agent, soap, liquid detergent, and spray. A cosmetic composition for preventing skin wrinkles, improving skin wrinkles, or reducing skin pores.
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| KR1020180063394A KR102628074B1 (en) | 2018-06-01 | 2018-06-01 | skin external application composition for anti-aging containing Luzula capitate extract |
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| KR1020180063394A KR102628074B1 (en) | 2018-06-01 | 2018-06-01 | skin external application composition for anti-aging containing Luzula capitate extract |
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2003201208A (en) * | 2001-12-28 | 2003-07-18 | Ichimaru Pharcos Co Ltd | Cosmetic composition |
| US20110311661A1 (en) * | 2003-11-18 | 2011-12-22 | Stephen Behr | Plant Extracts and Dermatological Uses Thereof |
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2003201208A (en) * | 2001-12-28 | 2003-07-18 | Ichimaru Pharcos Co Ltd | Cosmetic composition |
| US20110311661A1 (en) * | 2003-11-18 | 2011-12-22 | Stephen Behr | Plant Extracts and Dermatological Uses Thereof |
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| KR20190137330A (en) | 2019-12-11 |
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