KR20230010994A - Multifunctional cosmetic composition containing peat extracts and salts effective for skin wrinkle improvement, whitening, cell regeneration, and wound healing as an active ingredient - Google Patents

Multifunctional cosmetic composition containing peat extracts and salts effective for skin wrinkle improvement, whitening, cell regeneration, and wound healing as an active ingredient Download PDF

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KR20230010994A
KR20230010994A KR1020210091474A KR20210091474A KR20230010994A KR 20230010994 A KR20230010994 A KR 20230010994A KR 1020210091474 A KR1020210091474 A KR 1020210091474A KR 20210091474 A KR20210091474 A KR 20210091474A KR 20230010994 A KR20230010994 A KR 20230010994A
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extract
cosmetic composition
skin
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손진성
박상욱
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농업회사법인 제일인터내셔널주식회사
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    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00Cosmetics or similar toiletry preparations
    • A61K8/18Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/96Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
    • A61K8/97Cosmetics 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
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00Cosmetics or similar toiletry preparations
    • A61K8/18Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/19Cosmetics or similar toiletry preparations characterised by the composition containing inorganic ingredients
    • A61K8/20Halogens; Compounds thereof
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00Cosmetics or similar toiletry preparations
    • A61K8/18Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/30Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
    • A61K8/64Proteins; Peptides; Derivatives or degradation products thereof
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00Cosmetics or similar toiletry preparations
    • A61K8/18Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/96Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
    • A61K8/97Cosmetics 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
    • A61K8/9783Angiosperms [Magnoliophyta]
    • A61K8/9794Liliopsida [monocotyledons]
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00Cosmetics or similar toiletry preparations
    • A61K8/18Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/96Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
    • A61K8/98Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution of animal origin
    • A61K8/987Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution of animal origin of species other than mammals or birds
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61QSPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q19/00Preparations for care of the skin
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2800/00Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
    • A61K2800/40Chemical, physico-chemical or functional or structural properties of particular ingredients
    • A61K2800/59Mixtures
    • A61K2800/592Mixtures of compounds complementing their respective functions
    • A61K2800/5922At least two compounds being classified in the same subclass of A61K8/18

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  • Cosmetics (AREA)

Abstract

Provided is a multifunctional cosmetic composition containing a peat extract and salt effective for elasticity improvement, wrinkle improvement, whitening, anti-inflammatory, atopy, cell regeneration, and wound healing of skin as active ingredients comprising: the peat extract which is extracted with water of 65-85 ℃ as an extract solvent; and 5-15 parts by weight of the salt for 100 parts by weight of the peat extract as the active ingredients. The multifunctional cosmetic composition selectively further comprises: one or more ingredients selected from a group consisting of fermented protaetia brevitarsis seulensis larval peptide powder; fermented protaetia brevitarsis seulensis larval oil; a protaetia brevitarsis seulensis larval ethanol extract; and a kamchatka pleurospermum ethanol and acorus calamus ethanol extract.

Description

피부의 주름 개선, 미백, 세포 재생, 상처 치유에 유효한 피트 추출물과 소금을 활성성분으로 함유하는 다 기능성 화장품 조성물{Multifunctional cosmetic composition containing peat extracts and salts effective for skin wrinkle improvement, whitening, cell regeneration, and wound healing as an active ingredient}Multifunctional cosmetic composition containing peat extracts and salts effective for skin wrinkle improvement, whitening, cell regeneration, and wound healing healing as an active ingredient}

본 발명은 피트 추출물을 활성 성분으로 포함하여 피부 탄력 증진, 주름 개선, 피부 미백, 항염, 아토피, 피부 재생, 피부 상처 회복 효과가 우수한 다 기능성 화장품 조성물에 관한 것이다.The present invention relates to a multifunctional cosmetic composition containing pit extract as an active ingredient and having excellent skin elasticity enhancement, wrinkle improvement, skin whitening, anti-inflammatory, atopy, skin regeneration, and skin wound recovery effects.

피트는 죽은 나무·관목·이끼 등이 얕은 산성 수에서 썩은 다음 그 위에 쌓인 다른 나무 때문에 압축되어서 만들어지는 것으로서 이와 같은 피트의 효과는 여드름, 아토피, 건선 피부 등 피부 발진과 두피 처방에 좋은 효과가 있으며, 혈액 순환과 신진대사를 활발하게 하여 손, 발의 차가운 증상을 개선하는 효과가 확인되었으며, 특히 피부를 탄력적이게 하며, 보습 등 피부 개선에 효과가 우수한 것으로 알려져 있다.Pit is made by rotting dead trees, shrubs, moss, etc. in shallow acidic water and then compressing it with other trees piled on top of it. , The effect of improving cold symptoms of hands and feet by activating blood circulation and metabolism was confirmed, and in particular, it is known to be effective in improving skin elasticity and moisturizing.

또한, 미국 등록특허 6267962호(1997년 6월 30일 출원)에서는 곰팡이, 박테리아, 리케치아, 바이러스 감염, 건선(psoriasis), 알레르기 및 다른 피부염, 습진(eczema)에 의해 일어나는 상처 치료, 고통, 가려움증, 염증, 비정상적인 세포 증식 또는 감염의 치료에 사용할 수 있는 피트 또는 이의 유사물로부터 제조된 조성물을 개시하고 있다. In addition, US Patent No. 6267962 (filed on June 30, 1997) treats wounds caused by fungi, bacteria, rickettsia, viral infections, psoriasis, allergies and other dermatitis, eczema, pain, itching, A composition made from peat or the like that can be used for the treatment of inflammation, abnormal cell proliferation or infection is disclosed.

그러나 상술한 바와 같은 피트의 다양한 효과에도 피트를 주성분으로 하여 화장품 조성물을 상품화하기에는 그 활성을 약하여 한계를 가지고 있다. However, despite the various effects of pits as described above, commercialization of cosmetic compositions containing pits as a main component has limitations due to their weak activity.

1. 미국 등록특허 6267962호1. US Patent No. 6267962

따라서 본 발명이 이루고자 하는 과제는 피트 추출물에 그 활성 강화할 수 있는 천연물을 더 부가하여 피부 탄력 증진, 주름 개선, 피부 미백, 항염, 아토피, 피부 재생, 피부 상처 회복 효과가 우수한 다 기능성 화장품 조성물을 제공하는 것을 과제로 한다.Therefore, the problem to be achieved by the present invention is to provide a multifunctional cosmetic composition with excellent skin elasticity enhancement, wrinkle improvement, skin whitening, anti-inflammatory, atopy, skin regeneration, and skin wound recovery by adding a natural substance capable of enhancing its activity to a pit extract. make it a task to do

상기 기술적 과제를 달성하기 위하여, 본 발명은In order to achieve the above technical problem, the present invention

추출 용매를 65 내지 85℃의 물로 하여 추출한 피트 추출물; 및peat extract extracted with water at 65 to 85° C. as the extraction solvent; and

상기 피트 추출물 100중량부에 대하여 5 내지 15중량부의 소금을 활성성분으로 포함하는 것을 특징으로 하는 다 기능성 화장품 조성물을 제공한다.It provides a multi-functional cosmetic composition comprising 5 to 15 parts by weight of salt as an active ingredient based on 100 parts by weight of the pit extract.

상술한 바와 같은 본 발명에 따른 다 기능성 화장품 조성물은 발효 흰점박이꽃무지 유충 펩타이드 분말, 발효 흰점박이꽃무지 유충 오일, 흰점박이꽃무지 유충 에탄올 추출물, 왜우산풀 에탄올 및 창포 에탄올 추출물로 이루어진 군에서 선택되는 어느 하나 이상의 성분을 활성성분으로 더 포함하며, 그 함량은 상기 피트 추출물 100중량부에 대하여 5중량부인 것을 특징으로 한다.As described above, the multifunctional cosmetic composition according to the present invention is in the group consisting of fermented white-spotted radish larvae peptide powder, fermented white-spotted radish larvae oil, white-spotted radish larvae ethanol extract, Japanese umbrella plant ethanol, and calamus ethanol extract. It further includes any one or more selected ingredients as an active ingredient, and the content is characterized in that 5 parts by weight based on 100 parts by weight of the pit extract.

상술한 바와 같은 본 발명에 따른 다 기능성 화장품 조성물에 있어서,In the multifunctional cosmetic composition according to the present invention as described above,

상기 다 기능성은 피부 탄력 증진, 피부 미백, 피부 주름 방지, 항산화, 항염증, 항아토피, 피부 재생 및 피부 상처 치유에 대하여 현저한 효과를 보이는 것이다.The multifunctionality is to show remarkable effects on skin elasticity enhancement, skin whitening, skin wrinkle prevention, antioxidant, anti-inflammatory, anti-atopic, skin regeneration and skin wound healing.

본 발명에 의하면 소금, 흰점박이꽃무지 유충, 왜우산풀, 창포를 이용한 피트의 피부 활성 효과 강화를 통하여 피부 탄력 증진, 주름 개선, 피부 미백, 항염, 아토피, 피부 재생, 피부 상처 회복 효과를 동시에 발휘할 수 있는 다 기능성 화장품의 제조가 가능하다.According to the present invention, skin elasticity enhancement, wrinkle improvement, skin whitening, anti-inflammation, atopy, skin regeneration, and skin wound recovery effects are simultaneously enhanced by enhancing the skin activity effect of pits using salt, white-spotted flower larvae, Japanese umbrella grass, and calamus. It is possible to manufacture multi-functional cosmetics that can exert their full potential.

이하 본 발명을 실시하기 위한 구체적인 내용을 실시예 및 시험예를 통하여 상세하게 설명하기로 한다.Hereinafter, specific details for carrying out the present invention will be described in detail through examples and test examples.

본 발명은 피부 탄력 증진, 주름 개선, 피부 미백, 항염, 아토피, 피부 재생, 피부 상처 회복 효과가 우수한 다 기능성 화장품 조성물을 제공하기 위하여 추출 용매를 65 내지 85℃의 물로 하여 추출한 피트 추출물; 및 상기 피트 추출물 100중량부에 대하여 5 내지 15중량부의 소금을 활성성분으로 포함하는 것을 조성물을 제공한다.The present invention is a peat extract extracted with water at 65 to 85 ° C. as an extraction solvent to provide a multifunctional cosmetic composition with excellent skin elasticity enhancement, wrinkle improvement, skin whitening, anti-inflammatory, atopy, skin regeneration, and skin wound recovery effects; And it provides a composition comprising 5 to 15 parts by weight of salt as an active ingredient based on 100 parts by weight of the pit extract.

상술한 바와 같은 본 발명은 100℃의 열수가 아닌 65 내지 85℃의 물로 하여 추출하였을 때 피트 추출물의 피부에 대한 활성이 증가하고 또 피트 추출물 100중량부에 대하여 5 내지 15중량부의 소금을 용해하면 상승 작용에 의하여 현저한 효과 향상이 있음을 확인하여 완성한 것이다.As described above, the present invention increases the activity of the pit extract on the skin when extracted with water at 65 to 85 ° C instead of hot water at 100 ° C, and dissolves 5 to 15 parts by weight of salt with respect to 100 parts by weight of the pit extract It was completed by confirming that there is a significant effect improvement by synergistic action.

상기 피트 추출물 대비 소금의 함량이 5중량부 미만이면 상승효과가 나타나지 않으며, 15중량부를 초과하는 경우는 피부 자극 등에 문제가 있으며 상승효과가 발생하지 않아 첨가하는 의미가 없어진다.If the content of salt compared to the peat extract is less than 5 parts by weight, synergistic effect does not appear, and if it exceeds 15 parts by weight, there is a problem such as skin irritation and the synergistic effect does not occur, so the meaning of adding is lost.

또한, 상기 소금은 특별한 제한은 없으나 자연에서 제조한 천일염 내지 자염인 것이 바람직하며, 상기 천일염은 염전에서 바닷물을 증발시켜 만든 소금이며, 자염은 바닷물을 끓여서 만든 소금을 의미한다.In addition, the salt is not particularly limited, but it is preferable that it is natural natural salt or self-salt.

또한. 본 발명은 여기에 선택적으로 발효 흰점박이꽃무지 유충 펩타이드 분말, 발효 흰점박이꽃무지 유충 오일, 흰점박이꽃무지 유충 에탄올 추출물, 왜우산풀 에탄올 및 창포 에탄올 추출물로 이루어진 군에서 선택되는 어느 하나 이상의 성분을 활성성분으로 더 포함하는 것을 특징으로 하는 다 기능성 화장품 조성물을 제공하며, 이들은 피트 추출물 100중량부 대비 5중량부 포함되는 것이 바람직하며, 왜우산풀과 창포는 뿌리는 제거한 지상부를 건조하여 사용하며 에탄올을 이용한 추출법은 본 발명이 속하는 기술분야에 널리 알려진 것이라면 특별한 제한이 없으며, 흰점박이꽃무지 유충을 절식하여 내부의 이물질을 제거한 후에 건조하여 에탄올로 추출한 것을 의미한다.also. In the present invention, any one or more components selected from the group consisting of selectively fermented white-spotted flower radish larvae peptide powder, fermented white-spotted flower radish larvae oil, white-spotted flower radish larvae ethanol extract, Japanese umbrella plant ethanol, and calamus ethanol extract It provides a multi-functional cosmetic composition characterized by further comprising as an active ingredient, and it is preferable that they contain 5 parts by weight relative to 100 parts by weight of peat extract, and Japanese umbrella grass and calamus are used by drying the above-ground part from which the roots are removed, The extraction method using ethanol is not particularly limited as long as it is widely known in the art to which the present invention belongs, and means that the white-spotted flower larvae are fasted to remove foreign substances therein, dried, and extracted with ethanol.

또한, 발효 흰점박이꽃무지 유충 펩타이드 분말 및 발효 흰점박이꽃무지 유충 오일은 흰점박이꽃무지 유충을 유산균으로 발효를 한 후에 얻어지는 펩타이드 분말 내지 오일로서 충남 태안에 소재하는 에이치엠오건강드림영농조합법인에 의해 화장품 원료 등재한 물질로서, 본 발명에서도 이를 이용하며, 구체적인 제조방법은 대한민국 등록특허 제10-1938348오 개시되어 있는데, 그 방법은 (a) 굼벵이 또는 분쇄한 굼벵이를 물에 넣고 Bacillus subtilis var. natto 균주, Bacillus subtilis 균주 및Aspergillus kawachii 균주로 이루어진 군에서 선택되는 어느 하나 이상의 균주를 접종하여 발효시키는 단계; 및 (b) 상기 (a) 단계 완료 후 지방산 오일을 함유하는 상층부의 유상과 수용성 펩타이드를 함유하는 하층부의 수상을 분리하여 수득하는 단계를 포함하는 것을 특징으로 하는 굼벵이로부터 지방산 오일 및 수용성 펩타이드를 분리하는 방법으로 구성되어 있다.In addition, the fermented white-spotted radish larvae peptide powder and the fermented white-spotted radish larvae oil are peptide powders or oils obtained after fermenting the white-spotted radish larvae with lactic acid bacteria. As a substance registered as a cosmetic raw material by , it is also used in the present invention, and a specific manufacturing method is disclosed in Korean Registered Patent No. 10-1938348, the method of which is (a) put slugs or pulverized slugs in water and add Bacillus subtilis var. fermenting by inoculating one or more strains selected from the group consisting of natto strains, Bacillus subtilis strains, and Aspergillus kawachii strains; And (b) separating and obtaining the oil phase of the upper part containing fatty acid oil and the water phase of the lower part containing water-soluble peptide after completion of step (a). Separating fatty oil and water-soluble peptide from slugs, characterized in that It is composed in such a way that

본 발명에서의 화장품 조성물은 화장품 조성물에 통상적으로 이용되는 성분들을 포함할 수 있으며, 일 예로 항산화제, 안정화제, 용해화제, 비타민, 안료 및 향료와 같은 통상적인 보조제, 그리고 담체를 포함한다.The cosmetic composition of the present invention may include components commonly used in cosmetic compositions, and for example, antioxidants, stabilizers, solubilizers, vitamins, pigments, and flavoring agents, and carriers.

이와 같은 화장품 조성물은 당업계에서 통상적으로 제조되는 어떠한 제형으로도 제조될 수 있다. 일 구체 예로 고점도 유화제형, 저점도 유화제형 및 가용화 제형으로 이루어진 군에서 선택된 어느 하나의 제형일 수 있으나, 이에 제한되는 것은 아니다. 제조하고자 하는 제형에 따라 발명이 속하는 기술 분야에서 통상적으로 사용되는 화장품 조성물 배합 성분을 포함할 수 있다. 일 예로 용액, 현탁액, 유탁액, 페이스트, 겔, 크림, 로션, 파우더, 비누, 계면활성제-함유 클렌징, 오일, 분말 파운데이션, 유탁액 파운데이션, 왁스 파운데이션, 팩, 마사지크림 및 스프레이 등으로 제형화 될 수 있으나, 이에 한정되는 것은 아니다. 보다 상세하게는, 유연 화장수, 영양 화장수, 영양 크림, 마사지 크림, 에센스, 아이 크림, 선로션, 선크림, 메이크업 베이스, 비비크림, 클렌징크림, 클렌징폼, 클렌징 워터, 팩, 스틱상 제품, 밤(Balm) 타입 제품, 스프레이 또는 파우더의 제형으로 제조될 수 있다. Such a cosmetic composition may be prepared in any formulation conventionally prepared in the art. As a specific example, it may be any one formulation selected from the group consisting of a high-viscosity emulsifier type, a low-viscosity emulsifier type, and a solubilization formulation, but is not limited thereto. Depending on the formulation to be prepared, cosmetic composition ingredients commonly used in the technical field to which the invention pertains may be included. For example, solutions, suspensions, emulsions, pastes, gels, creams, lotions, powders, soaps, surfactant-containing cleansing, oils, powder foundations, emulsion foundations, wax foundations, packs, massage creams and sprays can be formulated. It may, but is not limited thereto. More specifically, softening lotion, nourishing lotion, nourishing cream, massage cream, essence, eye cream, sun lotion, sun cream, makeup base, BB cream, cleansing cream, cleansing foam, cleansing water, pack, stick product, balm ( Balm) type product, spray or powder formulation.

제조하고자 하는 제형에 따라 추가로 수상성분, 유상성분, 계면활성제, 보습제, 저급알코올, 점증제, 킬레이트제, 방부제, 향료 등을 선택하여 배합 첨가할 수 있다.Depending on the formulation to be prepared, an aqueous phase component, an oil phase component, a surfactant, a moisturizer, a lower alcohol, a thickener, a chelating agent, a preservative, a flavoring agent, etc. may be selected and added.

이하, 본 발명을 하기의 실시예 및 실험예에 의해 상세히 설명한다. 단, 하기 실시예 및 실험예는 본 발명을 예시하는 것일 뿐, 본 발명의 내용이 하기 실시예 및 실험예에 의해 한정되는 것은 아니다.Hereinafter, the present invention will be described in detail by the following Examples and Experimental Examples. However, the following Examples and Experimental Examples are only to illustrate the present invention, and the content of the present invention is not limited by the following Examples and Experimental Examples.

<< 실시예Example 1> 1>

피트 6kg을 75℃로 가온한 정제수 60ℓ와 혼합하여 3시간 동안 75℃를 유지하면서 추출을 하여 피트 추출물을 제조한 다음에 여기에 태안 천일염으로 만든 태현보호작업장의 RED SALT를 용해해 본 발명에 따른 조성물을 제조하였다.6 kg of peat was mixed with 60 liters of purified water heated to 75 ° C, extracted while maintaining 75 ° C for 3 hours to prepare a pit extract, and then dissolved RED SALT from Taehyeon Protection Works made from Taean sun salt to produce according to the present invention A composition was prepared.

여기서 RED SALT의 사용량은 피트 추출물 100중량부에 대하여 10중량부이었다.Here, the amount of RED SALT used was 10 parts by weight based on 100 parts by weight of the pit extract.

<< 실시예Example 2> 2>

실시예 1과 동일하며 다만 흰점박이꽃무지 유충 에탄올 추출물을 피트 추출물 100중량부에 대하여 5중량부 더 포함하여 제조한 것에 차이가 있다.It is the same as in Example 1, except that it was prepared by further including 5 parts by weight of the ethanol extract of the white-spotted flower larvae based on 100 parts by weight of the pit extract.

<< 실시예Example 3> 3>

실시예 1과 동일하며 다만 왜우산풀 에탄올 추출물을 피트 추출물 100중량부에 대하여 5중량부 더 포함하여 제조한 것에 차이가 있다.It is the same as Example 1, but there is a difference in that it was prepared by further including 5 parts by weight of the ethanol extract of Japanese umbrella plant relative to 100 parts by weight of the pit extract.

<< 실시예Example 4> 4>

실시예 1과 동일하며 다만 창포 에탄올 추출물을 피트 추출물 100중량부에 대하여 5중량부 더 포함하여 제조한 것에 차이가 있다.It is the same as in Example 1, except that the ethanol extract of Calamus was prepared by further including 5 parts by weight based on 100 parts by weight of pit extract.

<< 실시예Example 5> 5>

실시예 1과 동일하며 다만 흰점박이꽃무지 유충, 왜우산풀, 창포를 동일 중량비율로 혼합한 후에 제조한 에탄올 추출물을 피트 추출물 100중량부에 대하여 5중량부 더 포함하여 제조한 것에 차이가 있다.It is the same as in Example 1, except that the ethanol extract prepared after mixing the white-spotted flower larvae, Japanese umbrella plant, and calamus in the same weight ratio was prepared by further including 5 parts by weight based on 100 parts by weight of the peat extract. .

<< 실시예Example 6> 6>

실시예 1과 동일하며 다만 발효 흰점박이꽃무지 유충 펩타이드 분말을 피트 추출물 100중량부에 대하여 5중량부 더 포함하여 제조한 것에 차이가 있다.It is the same as in Example 1, except that it was prepared by further including 5 parts by weight of the fermented white-spotted radish larvae peptide powder based on 100 parts by weight of the pit extract.

상기 발효 흰점박이꽃무지 유충 펩타이드 분말은 충남 태안에 소재하는 에이치엠오건강드림영농조합법인으로부터 공급받아 사용하였다.The fermented white-spotted radish larvae peptide powder was supplied and used from HMO Health Dream Farming Association located in Taean, Chungcheongnam-do.

<< 실시예Example 7> 7>

실시예 1과 동일하며 다만 발효 흰점박이꽃무지 유충 오일을 피트 추출물 100중량부에 대하여 5중량부 더 포함하여 제조한 것에 차이가 있다.It is the same as in Example 1, except that it was prepared by further including 5 parts by weight of fermented white-spotted radish larvae oil based on 100 parts by weight of pit extract.

상기 발효 흰점박이꽃무지 유충 오일은 충남 태안에 소재하는 에이치엠오건강드림영농조합법인으로부터 공급받아 사용하였다.The fermented white-spotted flower radish larvae oil was supplied and used from HMO Health Dream Farming Association located in Taean, Chungcheongnam-do.

<< 비교예comparative example 1> 1>

피트 6kg을 75℃로 가온한 정제수 60ℓ와 혼합하여 3시간 동안 75℃를 유지하면서 추출을 하여 피트 추출물을 활성성분으로 포함하는 조성물을 제조하였다.6 kg of peat was mixed with 60 liters of purified water heated to 75° C. and extracted while maintaining the temperature at 75° C. for 3 hours to prepare a composition containing the peat extract as an active ingredient.

<< 비교예comparative example 2> 2>

피트 6kg을 100℃로 가온한 정제수 60ℓ와 혼합하여 3시간 동안 100℃를 유지하면서 추출을 하여 피트 추출물을 활성성분으로 포함하는 조성물을 제조하였다.6 kg of peat was mixed with 60 L of purified water heated to 100 ° C. and extracted while maintaining 100 ° C. for 3 hours to prepare a composition containing the peat extract as an active ingredient.

<< 시험예test example 1: One: ElastaseElastase 저해 활성을 통한 피부 탄력 증진 및 주름 방지 효능 평가> Evaluation of skin elasticity enhancement and anti-wrinkle efficacy through inhibitory activity>

실시예 및 비교예 조성물을 메탄올에 희석한 시험용액 40㎕에 50mM Tris-HCl buffer (pH 8.6)에 녹인 proporcine pancreas elastase (2.5 U/㎖) 용액 40㎕을 가한 후 기질로 50mM Tris-HCl buffer (pH 8.6)에 녹인 N-succinyl-(L-Ala)3-p-nitroanilide (0.5mg/㎖)을 80㎕ 첨가하여 혼합하였으며. 37℃에서 30분간 반응시킨 후 410nm에서 흡광도를 측정하였다. 양성 대조군으로는 oleanolic acid를 사용하여다. After adding 40 μl of a solution of proporcine pancreas elastase (2.5 U/ml) dissolved in 50 mM Tris-HCl buffer (pH 8.6) to 40 μl of the test solution diluted with the compositions of Examples and Comparative Examples in methanol, 50 mM Tris-HCl buffer ( 80 μl of N-succinyl-(L-Ala)3-p-nitroanilide (0.5 mg/ml) dissolved in pH 8.6) was added and mixed. After reacting at 37° C. for 30 minutes, absorbance was measured at 410 nm. As a positive control, oleanolic acid was used.

Elastase 저해활성은 하기 계산식과 같이 시료용액의 첨가구와 무첨가구의 흡광도 감소율로 계산하여 그 결과를 표 1에 나타냈다.Elastase inhibitory activity was calculated by the absorbance reduction rate of the sample solution added and unadded as in the following formula, and the results are shown in Table 1.

계산식: 저해율(%)= 1-(시료첨가군의 흡광도/시료무첨가군의 흡광도) x 100 Calculation formula: Inhibition rate (%) = 1-(absorbance of sample added group/absorbance of sample-free group) x 100

SampleSample Concentration (100 ㎍/㎖)Concentration (100 μg/mL) 실시예 1 Example 1 82.3±0.682.3±0.6 실시예 2 Example 2 85.9±0.785.9±0.7 실시예 3 Example 3 86.3±0.686.3±0.6 실시예 4 Example 4 86.2±0.586.2±0.5 실시예 5 Example 5 90.3±0.690.3±0.6 실시예 6Example 6 91.5±0.491.5±0.4 실시예 7Example 7 92.6±0.692.6±0.6 비교예 1Comparative Example 1 55.4±0.655.4±0.6 비교예 2Comparative Example 2 32.7±0.632.7±0.6 oleanolic acid oleanolic acid 70.6±0.570.6±0.5

인체의 중성구 과립구내에 존재하는 elastase는 동물 결합 조직의 불용성 탄성 섬유 단백질인 elastin을 분해시켜 피부의 진피조직의 그물망 구조 결합을 끊어줌으로써 피부 탄력 증진 감소 및 주름 생성의 주원인 효소로 알려져 있으며, 이의 활성을 억제하는 경우 피부 탄력 증진 유지 내지 회복, 피부 주름 생성을 방지할 수 있는데, 상기 표 1의 결과를 보면, 본 발명의 실시예 추출물 모두 양성 대조군보다 우수한 효과를 보여 소금의 상승 효과 등 그 효과의 현저성을 확인할 수 있었으며, 특히 실시예 5, 6, 7에서 매우 우수한 효과를 보였다. Elastase present in human neutrophil granulocytes degrades elastin, an insoluble elastic fiber protein of animal connective tissue, and breaks the network structure of the dermal tissue of the skin. In the case of suppression, it is possible to maintain or restore skin elasticity and prevent skin wrinkles. Looking at the results of Table 1 above, all the extracts of the examples of the present invention showed superior effects than the positive control group, and the synergistic effect of salt, etc. It was confirmed, and especially in Examples 5, 6, and 7, very good effects were shown.

또한, 비교예를 보면 열수로 추출한 비교예 2에서 비교예 1보다 효과가 떨어지는 것으로 보아 피트는 100℃보다 낮은 온도에서 추출하는 것은 효율적임을 알 수 있었다.In addition, looking at Comparative Example, it was found that extracting pits at a temperature lower than 100 ° C. was effective, as the effect was lower than that of Comparative Example 1 in Comparative Example 2 extracted with hot water.

<< 시험예test example 2: 미백 효능 평가> 2: Evaluation of whitening efficacy>

1) One) tyrosinasetyrosinase 저해 활성 측정 Inhibition activity measurement

0.175M sodium phosphate buffer(pH 6.8) 0.5㎖에 기질액인 10mM L-DOPA 0.2㎖ 과 시료용액 0.1㎖을 96 well plate에 넣고 혼합한 혼합액에 mushroom tyrosinase (110 U/㎖) 0.2㎖을 첨가하여 25℃에서 2분간 반응시켜 반응액 중에 생성된 DOPA chrome을 475nm에서 측정하였으며, 양성 대조군으로 Arbutin을 사용하였다. 0.175M sodium phosphate buffer (pH 6.8) 0.2ml of 10mM L-DOPA and 0.1ml of sample solution were added to 0.5ml of 0.175M sodium phosphate buffer (pH 6.8) in a 96 well plate. DOPA chrome generated in the reaction solution by reacting at ℃ for 2 minutes was measured at 475 nm, and Arbutin was used as a positive control.

Tyrosinase 저해활성은 하기 계산식과 같이 시료용액의 첨가구와 무첨가구의 흡광도 감소율로 계산하여 그 결과를 표 2에 나타냈다.Tyrosinase inhibitory activity was calculated by the absorbance reduction rate of the sample solution added and unadded as in the following formula, and the results are shown in Table 2.

계산식: 저해율(%)= 1-(시료첨가군의 흡광도/시료무첨가군의 흡광도) x 100Calculation formula: Inhibition rate (%) = 1-(absorbance of sample added group/absorbance of sample-free group) x 100

SampleSample Concentration (100 ㎍/㎖)Concentration (100 μg/ml) 실시예 1 Example 1 67.8±0.367.8±0.3 실시예 2 Example 2 76.5±0.576.5±0.5 실시예 3 Example 3 75.9±0.675.9±0.6 실시예 4 Example 4 74.2±0.574.2±0.5 실시예 5 Example 5 81.9±0.681.9±0.6 실시예 6Example 6 82.7±0.582.7±0.5 실시예 7Example 7 82.4±0.682.4±0.6 비교예 1Comparative Example 1 42.5±0.842.5±0.8 비교예 2Comparative Example 2 29.5±0.829.5±0.8 ArbutinArbutin 55.8±0.855.8±0.8

tyrosinase는 피부 멜라닌 생성에 있어서 매우 중요한 역할을 하고 있으며, melanosome 내에서 tyrosine을 산화시켜 DOPA를 만드는 tyrosine hydroxylase로 DOPA를 산화시켜 DOPA quinone을 만드는 DOPA oxidase로서 작용하여 멜라닌 중합체를 합성하는데 중요한 효소로 작용하며, 이를 억제하면 멜라닌의 생성의 억제되어 피부 미백 효과를 나타내게 되는데, 상기 표 2의 결과를 보면, 본 발명의 실시예 추출물 모두 양성 대조군보다 우수한 효과를 보여 소금을 통한 상승효과 등 그 효과의 현저성을 확인할 수 있었으며, 특히 실시예 5, 6, 7에서 매우 우수한 효과를 보였다.Tyrosinase plays a very important role in the production of melanin in the skin. It is a tyrosine hydroxylase that oxidizes tyrosine in the melanosome to make DOPA. It oxidizes DOPA to make DOPA quinone. , If this is suppressed, the production of melanin is suppressed to exhibit a skin whitening effect. Looking at the results of Table 2, all the extracts of the examples of the present invention showed a superior effect than the positive control group, and the synergistic effect through salt It was confirmed, and in particular, Examples 5, 6, and 7 showed very good effects.

또한, 비교예를 보면 열수로 추출한 비교예 2에서 비교예 1보다 효과가 떨어지는 것으로 보아 피트는 100℃보다 낮은 온도에서 추출하는 것은 효율적임을 알 수 있었다.In addition, looking at Comparative Example, it was found that extracting pits at a temperature lower than 100 ° C. was effective, as the effect was lower than that of Comparative Example 1 in Comparative Example 2 extracted with hot water.

2) melanin 생합성 저해 활성 측정 2) Measurement of melanin biosynthesis inhibitory activity

melanoma cell line인 B16F10 cell을 한국 세포주 은행 (Korean Cell Line Bank)으로부터 구입하였으며, 1% antibiotic (Gibco, USA)과 10% fetal bovine serum (FBS; Gibco, Grand Island, USA)이 함유된 Dulbecco's modified Eagle's medium (DMEM) 배지를 사용하여 37℃, 5% CO2 incubator에서 배양하였으며, 2일에 한번씩 계대 배양을 실시하였다. B16F10 cells, a melanoma cell line, were purchased from the Korean Cell Line Bank, Dulbecco's modified Eagle's supplemented with 1% antibiotic (Gibco, USA) and 10% fetal bovine serum (FBS; Gibco, Grand Island, USA). It was cultured in a 37°C, 5% CO 2 incubator using a medium (DMEM) medium, and subculture was performed once every 2 days.

DMEM 배지로 배양된 B16F10 세포를 24 well plate에 2.0 X 104 cells/well로 접종한 후 37℃, 5% CO2 incubator에서 24시간 배양 후 시료와 혼합하였다.B16F10 cells cultured in DMEM medium were inoculated in a 24 well plate at 2.0 X 10 4 cells/well, and then cultured for 24 hours in a 37°C, 5% CO 2 incubator and mixed with the sample.

시료와 cell에 자극하기 위하여 자극제인 α-MSH를 200nM로 혼합하여 첨가한 후 37℃, 5% CO2 incubator에서 48시간 동안 배양한 후에 배양액을 제거한 후 인산완충액(pH 7.4)으로 각 well을 세척하였다.To stimulate the samples and cells, α-MSH, a stimulant, was mixed and added at 200 nM, cultured for 48 hours in an incubator at 37°C, 5% CO 2 , then the culture medium was removed, and each well was washed with phosphate buffer (pH 7.4). did

이어서 1N NaOH를 100㎕씩 가하여 cell을 떼어낸 후 1.5㎖ tube에 옮겨준다. 80℃에서 1시간 반응시킨 후 분광광도계 405nm에서 흡광도를 측정하였으며, melanin 생합성 저해는 시료 용액의 첨가구와 무첨가구의 흡광도 감소율로 계산하였으며, 그 결과는 표 3에 나타냈다.Subsequently, 100 μl of 1N NaOH was added to detach the cells, and then transferred to a 1.5 ml tube. After reacting at 80 ℃ for 1 hour, the absorbance was measured at 405 nm with a spectrophotometer, and melanin biosynthesis inhibition was calculated by the absorbance decrease rate of the sample solution added and unadded, and the results are shown in Table 3.

samplesample Melanin contents (%)Melanin contents (%) α-MSH(-)α-MSH(-) -- α-MSH(+, 200nm)α-MSH (+, 200 nm) 100100 α-MSH(+, 200nM) + 실시예 1 α-MSH (+, 200 nM) + Example 1 42.8 ± 1.242.8 ± 1.2 α-MSH(+, 200nM) + 실시예 2 α-MSH (+, 200 nM) + Example 2 35.9 ± 1.535.9±1.5 α-MSH(+, 200nM) + 실시예 3 α-MSH (+, 200 nM) + Example 3 35.1 ± 1.135.1 ± 1.1 α-MSH(+, 200nM) + 실시예 4 α-MSH (+, 200 nM) + Example 4 34.5 ± 1.334.5 ± 1.3 α-MSH(+, 200nM) + 실시예 5 α-MSH (+, 200 nM) + Example 5 26.1 ± 1.226.1 ± 1.2 α-MSH(+, 200nM) + 실시예 6α-MSH (+, 200 nM) + Example 6 24.8 ± 1.124.8 ± 1.1 α-MSH(+, 200nM) + 실시예 7α-MSH (+, 200 nM) + Example 7 25.1 ± 1.225.1 ± 1.2 α-MSH(+, 200nM) + 비교예 1α-MSH (+, 200 nM) + Comparative Example 1 66.2 ± 1.366.2 ± 1.3 α-MSH(+, 200nM) + 비교예 2α-MSH (+, 200 nM) + Comparative Example 2 77.8 ± 1.377.8 ± 1.3

상기 표 3을 보면 α-MSH 단독 처리군을 통해 α-MSH에 의해 멜라닌 합성이 유도됨을 확인할 수 있었으며, α-MSH 단독 처리군에 비교하여 본 발명의 실시예 추출물 모두 우수한 효과를 보여 소금을 통한 상승효과 등 그 효과의 현저성을 확인할 수 있었으며, 특히 실시예 5, 6, 7에서 매우 우수한 효과를 보였다.Looking at Table 3, it was confirmed that melanin synthesis was induced by α-MSH through the α-MSH alone treatment group, and all of the example extracts of the present invention showed excellent effects compared to the α-MSH alone treatment group. Significance of the effect, such as synergistic effect, was confirmed, and particularly excellent effects were shown in Examples 5, 6, and 7.

<< 시험예test example 3: Nitric oxide (NO) 생성 억제 활성을 통한 항염증 효능 평가> 3: Evaluation of anti-inflammatory efficacy through inhibition of nitric oxide (NO) production>

Murine macrophage cell line인 RAW 264.7 cell을 한국 세포주 은행 (Korean Cell Line Bank)으로부터 구입하였으며, 1% antibiotic (Gibco, USA)과 10% fetal bovine serum (FBS; Gibco, Grand Island, USA)이 함유된 Dulbecco's modified Eagle's medium (DMEM) 배지를 사용하여 37℃, 5% CO2 incubator에서 배양하였으며, 2일에 한번씩 계대 배양을 실시하였다. RAW 264.7 cells, a murine macrophage cell line, were purchased from the Korean Cell Line Bank and were supplemented with 1% antibiotic (Gibco, USA) and 10% fetal bovine serum (FBS; Gibco, Grand Island, USA) from Dulbecco's. It was cultured in a 37°C, 5% CO 2 incubator using modified Eagle's medium (DMEM) medium, and subculture was performed once every 2 days.

RAW 264.7 cell를 10% FBS가 첨가된 DMEM 배지를 이용하여 2.0 × 105 cells/㎖로 24 well plate에 넣고 18시간 배양하였으며, 준비된 시료에 LPS를 1ug/㎖의 농도로 혼합하여 cell에 동시에 처리하여 37℃, 5% CO2 incubator에서 24시간 배양하였다.RAW 264.7 cells were placed in a 24 well plate at 2.0 × 10 5 cells/ml using DMEM medium supplemented with 10% FBS and cultured for 18 hours. LPS was mixed with the prepared sample at a concentration of 1 ug/ml and the cells were simultaneously treated. and cultured for 24 hours in a 37°C, 5% CO 2 incubator.

이어서 세포배양 상등액 100㎕와 Griess 시약 100㎕를 혼합하여 96 well plate에서 10분 동안 반응시킨 후 540nm에서 흡광도를 측정하였다. Subsequently, 100 μl of the cell culture supernatant and 100 μl of Griess reagent were mixed and reacted in a 96 well plate for 10 minutes, and then absorbance was measured at 540 nm.

생성된 NO의 양은 Griess 시약 [1% (w/v) sulfanilamide, 0.1% (w/v) naphylethylenediamine in 2.5% (v/v) phosphoric acid]을 이용하여 세포배양액 중에 존재하는 NO2 -의 형태로 측정하고 sodium nitrite (NaNO2)를 standard로 사용하여 확인할 수 있었으며, 그 결과는 표 4에 나타냈으며, 양성 대조군으로 대조군인 2-amino- 4-methylpyridine을 사용하였다.The amount of NO produced was determined in the form of NO 2 - present in the cell culture medium using Griess reagent [1% (w/v) sulfanilamide, 0.1% (w/v) naphylethylenediamine in 2.5% (v/v) phosphoric acid]. It was measured and confirmed using sodium nitrite (NaNO 2 ) as a standard. The results are shown in Table 4, and 2-amino-4-methylpyridine, a control group, was used as a positive control.

SampleSample NO production (%)NO production (%) LPS (-)LPS (-) -- LPS (+, 1㎍/㎖ )LPS (+, 1 μg/ml) 100100 2-amino-4-methylpyridine 2-amino-4-methylpyridine 36.8 ± 1.936.8 ± 1.9 LPS (1㎍/㎖) + 실시예 1(1000㎍/㎖)LPS (1 μg/ml) + Example 1 (1000 μg/ml) 29.9 ± 1.329.9 ± 1.3 LPS (1㎍/㎖) + 실시예 2(1000㎍/㎖)LPS (1 μg/ml) + Example 2 (1000 μg/ml) 20.7 ± 1.920.7 ± 1.9 LPS (1㎍/㎖) + 실시예 3(1000㎍/㎖)LPS (1 μg/ml) + Example 3 (1000 μg/ml) 20.9 ± 1.420.9 ± 1.4 LPS (1㎍/㎖) + 실시예 4(1000㎍/㎖)LPS (1 μg/ml) + Example 4 (1000 μg/ml) 20.9 ± 1.220.9 ± 1.2 LPS (1㎍/㎖) + 실시예 5(1000㎍/㎖)LPS (1 μg/ml) + Example 5 (1000 μg/ml) 17.1 ± 1.317.1 ± 1.3 LPS (1㎍/㎖) + 실시예 6(1000㎍/㎖)LPS (1 μg/ml) + Example 6 (1000 μg/ml) 15.7 ± 1.215.7±1.2 LPS (1㎍/㎖) + 실시예 7(1000㎍/㎖)LPS (1 μg/ml) + Example 7 (1000 μg/ml) 16.2 ± 1.316.2 ± 1.3 LPS (1㎍/㎖) + 비교예 1(1000㎍/㎖)LPS (1 μg/ml) + Comparative Example 1 (1000 μg/ml) 55.7 ± 1.955.7 ± 1.9 LPS (1㎍/㎖) + 비교예 2(1000㎍/㎖)LPS (1 μg/ml) + Comparative Example 2 (1000 μg/ml) 71.2 ± 1.971.2 ± 1.9

상기 표 4의 결과를 보면, 본 발명의 실시예 추출물 모두 양성 대조군보다 우수한 효과를 보여 소금을 통한 상승효과 등 그 효과의 현저성을 확인할 수 있었으며, 특히 실시예 5, 6, 7에서 매우 우수한 효과를 보였다.Looking at the results of Table 4, all of the extracts of the examples of the present invention showed a superior effect than the positive control group, and it was confirmed that the effect such as the synergistic effect through salt was remarkable, especially in Examples 5, 6, and 7. showed

<< 시힘예Sihimye 4: 염증성 4: inflammatory chemokineschemokines ( ( TARCTARC , , MDCMDC ) 생성 억제 활성을 통한 ) through production inhibition activity 항아토피anti-atopy 효능 평가> Efficacy evaluation>

Human keratinocyte cell line인 HaCaT cell을 한국 세포주 은행으로부터 구입하였으며, 1% antibiotic 과 10% fetal bovine serum 이 함유된 Dulbecco's modified Eagle's medium 배지를 사용하여 37℃, 5% CO2 incubator에서 배양하였으며, 3일에 한번씩 계대 배양을 실시하였다.HaCaT cells, a human keratinocyte cell line, were purchased from the Korean Cell Line Bank and cultured in a 37°C, 5% CO 2 incubator using Dulbecco's modified Eagle's medium containing 1% antibiotic and 10% fetal bovine serum. Subculture was performed once.

HaCaT cell을 3.0 * 105 cells/㎖로 24 well plate에 18시간 전 배양하였으며, 준비된 시료에 IFN-γ 와 TNF-α를 각각 10ng/㎖ 농도로 혼합하여 cell에 동시에 처리하여 37℃, 5% CO2 incubator에서 24시간 배양하였다. HaCaT cells 3.0 * 10 5 cells / ㎖ was cultured in a 24 well plate for 18 hours before, and IFN-γ and TNF-α were mixed with the prepared sample at a concentration of 10 ng / ㎖, respectively, and the cells were simultaneously treated at 37 ℃, 5% CO 2 incubator incubated for 24 hours.

이후 배양액을 1.5㎖ tube에 옮겨 원심 분리하여 얻어진 상층액의 chemonkines 합성량을 측정하였 그 결과를 표 5에 나타냈으며, 모든 시료는 정량 전까지 냉동보관 하엿으며, 염증성 chemokines는 Human enzyme-linked immnunosorbent assay (ELISA) kit (R&D Systems)를 이용하여 정량하였으며 standard에 대한 표준곡선의 r 2 값은 0.99 이상이었다.Subsequently, the culture medium was transferred to a 1.5 ml tube and centrifuged to measure the amount of chemonkines synthesized in the supernatant obtained. ELISA) kit (R&D Systems), and the r 2 value of the standard curve for the standard was 0.99 or more.

SampleSample TARC production (%)TARC production (%) -- 00 TNF-α (10ng/㎖) + IFN-γ (10ng/㎖)TNF-α (10ng/ml) + IFN-γ (10ng/ml) 100100 TNF-α (10ng/㎖) + IFN-γ (10ng/㎖) +
실시예 1(1000㎍/㎖)
TNF-α (10ng/ml) + IFN-γ (10ng/ml) +
Example 1 (1000 μg/ml)
35.1 ± 1.335.1 ± 1.3
TNF-α (10ng/㎖) + IFN-γ (10ng/㎖) +
실시예 2(1000㎍/㎖)
TNF-α (10ng/ml) + IFN-γ (10ng/ml) +
Example 2 (1000 μg/ml)
27.9 ± 2.127.9 ± 2.1
TNF-α (10ng/㎖) + IFN-γ (10ng/㎖) +
실시예 3(1000㎍/㎖)
TNF-α (10ng/ml) + IFN-γ (10ng/ml) +
Example 3 (1000 μg/ml)
28.3 ± 1.628.3 ± 1.6
TNF-α (10ng/㎖) + IFN-γ (10ng/㎖) +
실시예 4(1000㎍/㎖)
TNF-α (10ng/ml) + IFN-γ (10ng/ml) +
Example 4 (1000 μg/ml)
27.6 ± 1.527.6±1.5
TNF-α (10ng/㎖) + IFN-γ (10ng/㎖) +
실시예 5(1000㎍/㎖)
TNF-α (10ng/ml) + IFN-γ (10ng/ml) +
Example 5 (1000 μg/ml)
20.4 ± 1.620.4 ± 1.6
TNF-α (10ng/㎖) + IFN-γ (10ng/㎖) +
실시예 6(1000㎍/㎖)
TNF-α (10ng/ml) + IFN-γ (10ng/ml) +
Example 6 (1000 μg/ml)
19.5 ± 1.719.5 ± 1.7
TNF-α (10ng/㎖) + IFN-γ (10ng/㎖) +
실시예 7(1000㎍/㎖)
TNF-α (10ng/ml) + IFN-γ (10ng/ml) +
Example 7 (1000 μg/ml)
19.6 ± 1.319.6 ± 1.3
TNF-α (10ng/㎖) + IFN-γ (10ng/㎖) +
비교예 1(1000㎍/㎖)
TNF-α (10ng/ml) + IFN-γ (10ng/ml) +
Comparative Example 1 (1000 μg/ml)
67.1 ± 1.267.1 ± 1.2
TNF-α (10ng/㎖) + IFN-γ (10ng/㎖) +
비교예 2(1000㎍/㎖)
TNF-α (10ng/ml) + IFN-γ (10ng/ml) +
Comparative Example 2 (1000 μg/ml)
79.2 ± 1.279.2 ± 1.2

아토피 피부염 환자에서 TARC의 발현이 증가하는 것으로 알려져 있는데, 상기 표 5의 결과를 보면, TNF-α, IFN-γ 단독 처리군을 통해 TARC의 발현이 유도됨을 확인하였고, 실시예의 시료로 처리하였을 때 TNF-α, IFN-γ 단독 처리군과 비교하여 TARC의 발현에 대하여 높은 억제율을 보이는 것을 확인할 수 있었으며, 특히 실시예 5, 6, 7에서 매우 우수한 효과를 보였다.It is known that the expression of TARC is increased in patients with atopic dermatitis. Looking at the results of Table 5 above, it was confirmed that the expression of TARC was induced through the TNF-α and IFN-γ alone treatment group, and when treated with the sample of Example Compared to the TNF-α and IFN-γ treatment groups, it was confirmed that the expression of TARC was highly inhibited, and in particular, examples 5, 6, and 7 showed excellent effects.

<< 시힘예Sihimye 5: fibroblast 이동성 평가를 통한 피부 세포 재생 및 상처 치유 효과 평가> 5: Evaluation of skin cell regeneration and wound healing effects through fibroblast mobility evaluation>

Human dermal fibroblast cell을 한국 세포주 은행으로부터 구입하였으며, 1% antibiotic 과 10% fetal bovine serum 이 함유된 Dulbecco's modified Eagle's medium 배지를 사용하여 37℃, 5% CO2 incubator에서 배양하였으며, 3, 4일에 한번씩 계대 배양을 실시하였다. Human dermal fibroblast cells were purchased from the Korea Cell Line Bank and cultured in a 37°C, 5% CO 2 incubator using Dulbecco's modified Eagle's medium containing 1% antibiotic and 10% fetal bovine serum, and cultured once every 3 or 4 days. Subculture was performed.

fibroblast의 이동성 평가를 위해 CytoSelectTM 24-Well Wound Healing Assay kit (CELL BIOLABS INC. USA)를 사용하였다. 멸균된 포셉을 이용하여 wound field inserts를 24 well plate의 각 well에 잘 고정시킨 후 fibroblast를 24well plate에 각각 8.0x104 cells/well, 3.0x105 cells/well로 접종하였다. To evaluate the mobility of fibroblast, CytoSelect TM 24-Well Wound Healing Assay kit (CELL BIOLABS INC. USA) was used. After the wound field inserts were well fixed to each well of a 24-well plate using sterilized forceps, fibroblasts were inoculated into the 24-well plate at 8.0x10 4 cells/well and 3.0x10 5 cells/well, respectively.

이어서 37℃, 5% CO2 incubator에서 18시간 전배양 한 후 시료를 농도별로 준비한다. 멸균된 포셉을 이용하여 wound field insert를 제거한 후 배양액을 제거하고 PBS로 2번 세척한 다음 준비된 시료인 실시예 1의 조성물을 각 well에 처리한 후 37℃, 5% CO2 incubator에서 2시간 동안 배양한 후 배양액을 제거한 다음 PBS로 세척 후 시료가 들어있지 않은 배양액으로 교체하였다.After 18 hours of pre-incubation in a 37°C, 5% CO 2 incubator, samples are prepared by concentration. After removing the wound field insert using sterilized forceps, the culture medium was removed, washed twice with PBS, and then treated with the prepared sample, the composition of Example 1, in each well, and then incubated at 37°C, 5% CO 2 incubator for 2 hours. After culturing, the culture medium was removed, washed with PBS, and replaced with a culture medium containing no sample.

그리고 37℃, 5% CO2 incubator에서 24시간, 48시간, 72시간 동안 시간별로 cell 상태를 확인하고, 배양을 멈춰야 하는 시간 때에 배양액을 제거한 후 cell의 잔여물이 남지 않도록 PBS로 2번 세척한 다음 각 well에 cell stain solution을 250㎕ 씩 넣은 후 실온에서 15분 동안 반응시키고 다시 PBS로 3번 세척한 다음 실온에서 건조한 plate를 가지고 현미경으로 cell의 이동성 상태를 확인하였다.Then, check the cell status hourly for 24 hours, 48 hours, and 72 hours in a 37℃, 5% CO 2 incubator, remove the culture medium at the time to stop the culture, and wash twice with PBS so that no cell residue remains. Next, 250 μl of cell stain solution was added to each well, reacted at room temperature for 15 minutes, washed again with PBS three times, and the cell mobility was checked under a microscope with the plate dried at room temperature.

피부는 외부환경에 대한 일차 장벽으로 개체를 보호하는 주된 역할을 담당한다. fibroblast는 피부 내 진피 층 건조중량의 70%를 차지하는 collagen을 생성하며, 진피층을 이루는 주요 세포이다. 결손 된 피부의 진피층이 재생되기 위하여 fibroblast의 재생이 가장 중요한데, 실시예 추출물이fibroblast에 대한 상처치유에 영향을 미치는지 확인해 보기 위해 wound healing assay를 수행하였으며, 아무 처리 하지 않은 control군과 실시예 1 추출물을 각각 6.25㎍/㎖씩 처리한 세포군의 증식 정도를 관찰하여 그 결과를 표 6에 나타냈다. The skin plays a major role in protecting the individual as the primary barrier to the external environment. Fibroblasts produce collagen, which accounts for 70% of the dry weight of the dermal layer in the skin, and is the main cell constituting the dermal layer. In order to regenerate the dermal layer of the damaged skin, the regeneration of fibroblast is the most important, and a wound healing assay was performed to check whether the Example extracts affect wound healing on fibroblast. The untreated control group and Example 1 extract The degree of proliferation of each cell group treated with 6.25 μg/ml was observed and the results are shown in Table 6.

fibroblastfibroblast 시료sample N.CN.C. 시간hour 48시간 48 hours 72시간72 hours

Figure pat00001

Figure pat00001

Figure pat00002



Figure pat00002


시료sample 실시예 1 조성물Example 1 composition 시긴Sigyn 48시간48 hours 72시간72 hours
Figure pat00003


Figure pat00003


Figure pat00004


Figure pat00004

상기 표 6을 보면 48시간과 72시간을 배양하여 확인한 결과 실시예 1 추출물로 처리한 세포에서 시간이 지날수록 negative control보다 세포의 이동과 증식의 속도가 빠른 것으로 관찰되어 피부 세포 재생 및 상처 치유에 현저한 효과를 확인한 수 있었다.Looking at Table 6, as a result of culturing for 48 hours and 72 hours, it was observed that the speed of cell migration and proliferation was faster than that of the negative control over time in the cells treated with the extract of Example 1. A significant effect could be confirmed.

이상에서 살펴본 바와 같이 본 발명에 의하면 소금, 흰점박이꽃무지 유충, 왜우산풀, 창포를 이용한 피트의 피부 활성 효과 강화를 통하여 피부 탄력 증진, 주름 개선, 피부 미백, 항염, 아토피, 피부 재생, 피부 상처 회복 효과를 동시에 발휘할 수 있는 다 기능성 화장품의 제조가 가능하다.As described above, according to the present invention, skin elasticity enhancement, wrinkle improvement, skin whitening, anti-inflammation, atopy, skin regeneration, skin regeneration, and It is possible to manufacture multi-functional cosmetics that can simultaneously exhibit wound healing effects.

Claims (4)

추출 용매를 65 내지 85℃의 물로 하여 추출한 피트 추출물; 및
상기 피트 추출물 100중량부에 대하여 5 내지 15중량부의 소금을 활성성분으로 포함하는 것을 특징으로 하는 다 기능성 화장품 조성물.
peat extract extracted with water at 65 to 85° C. as the extraction solvent; and
A multifunctional cosmetic composition comprising 5 to 15 parts by weight of salt as an active ingredient based on 100 parts by weight of the pit extract.
제 1항에 있어서,
발효 흰점박이꽃무지 유충 펩타이드 분말, 발효 흰점박이꽃무지 유충 오일, 흰점박이꽃무지 유충 에탄올 추출물, 왜우산풀 에탄올 및 창포 에탄올 추출물로 이루어진 군에서 선택되는 어느 하나 이상의 성분을 활성성분으로 더 포함하는 것을 특징으로 하는 다 기능성 화장품 조성물.
According to claim 1,
Further containing as an active ingredient at least one component selected from the group consisting of fermented white-spotted radish larvae peptide powder, fermented white-spotted radish larvae oil, white-spotted radish larvae ethanol extract, Japanese umbrella grass ethanol, and calamus ethanol extract A multifunctional cosmetic composition, characterized in that.
제 2항에서 있어서,
상기 흰점박이꽃무지 유충 에탄올 추출물, 왜우산풀 에탄올 및 창포 에탄올 추출물로 이루어진 군에서 선택되는 어느 하나 이상의 성분이 상기 피트 추출물 100중량부에 대하여 5중량부 포함되는 것을 특징으로 하는 다 기능성 화장품 조성물.
In claim 2,
A multifunctional cosmetic composition, characterized in that 5 parts by weight of at least one component selected from the group consisting of the ethanol extract of the white-spotted flower radish larva, ethanol extract of Japanese umbrella plant, and ethanol extract of calamus is included based on 100 parts by weight of the pit extract.
전술한 항 중 어느 한 항에 있어서,
상기 다 기능성이 피부 탄력 증진, 피부 미백, 피부 주름 방지, 항산화, 항염증, 항아토피, 피부 재생 및 피부 상처 치유인 것을 특징으로 하는 다 기능성 화장품 조성물.
According to any one of the preceding claims,
A multifunctional cosmetic composition, characterized in that the multifunctionality is skin elasticity enhancement, skin whitening, skin wrinkle prevention, antioxidant, anti-inflammatory, anti-atopic, skin regeneration and skin wound healing.
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