KR20220152865A - Rigida pine bark extract with improved stability encapsulated nanoparticles and manufacturing method, cosmetic composition comprising the same - Google Patents
Rigida pine bark extract with improved stability encapsulated nanoparticles and manufacturing method, cosmetic composition comprising the same Download PDFInfo
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- KR20220152865A KR20220152865A KR1020210060332A KR20210060332A KR20220152865A KR 20220152865 A KR20220152865 A KR 20220152865A KR 1020210060332 A KR1020210060332 A KR 1020210060332A KR 20210060332 A KR20210060332 A KR 20210060332A KR 20220152865 A KR20220152865 A KR 20220152865A
- Authority
- KR
- South Korea
- Prior art keywords
- dermatitis
- skin
- cosmetic composition
- bark extract
- rigida
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- 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
- A61K8/9755—Gymnosperms [Coniferophyta]
- A61K8/9767—Pinaceae [Pine family], e.g. pine or cedar
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- A—HUMAN NECESSITIES
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- A61K8/02—Cosmetics or similar toiletry preparations characterised by special physical form
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- A—HUMAN NECESSITIES
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- A—HUMAN NECESSITIES
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- A—HUMAN NECESSITIES
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Abstract
Description
본 발명은 안정성이 향상된 리기다 소나무 껍질 추출물 봉입 나노 입자 및 제조방법, 이를 함유하는 화장료 조성물에 관한 것이다.The present invention relates to Rigida pine bark extract-encapsulated nanoparticles with improved stability, a manufacturing method, and a cosmetic composition containing the same.
아토피 피부염(Atopic dermatitis)은 만성적이고 재발성인 염증성 피부질환으로, 건조하고 각질이 많은 비늘피부, 염증, 증가된 피부 투과성과 일반적이고 무해한 인자에 대해 민감하게 반응하는 증상을 보이며, 표면 감염에 취약하며, 심각한 발작과 가려움을 나타내는 염증성 피부염의 일종이다.Atopic dermatitis is a chronic, recurrent inflammatory skin disease characterized by dry, flaky, scaly skin, inflammation, increased skin permeability, and sensitivity to common and harmless factors, and is susceptible to superficial infection. , a type of inflammatory dermatitis that presents with severe seizures and itching.
글로벌 아토피 피부염 환자수는 1.38억명이며 매년 높은 속도로 증가하고 있다. 전세계적으로 아토피 피부염 뿐만 아니라 전반적인 피부 염증성 질환은 발생원인이 매우 광범위할 뿐 아니라, 원인이 다양하여 치료에 어려움이 있었다. 현재는, 피부 염증으로 인한 가려움증 및 피부 손상을 개선 또는 예방하기 위해, 스테로이드제를 일반적으로 사용하고 있다. 그러나, 스테로이드제는 증상을 일시적으로 완화시키는 스테로이드 제제 이나 이는 피부를 얇게 하고 궁극적으로 증상을 더 악화시키는 부작용이 있는 문제점이 있었다.The number of global atopic dermatitis patients is 138 million and is increasing at a high rate every year. Globally, not only atopic dermatitis but also overall skin inflammatory diseases have a very wide range of causes and are difficult to treat due to various causes. Currently, in order to improve or prevent itching and skin damage caused by skin inflammation, steroids are generally used. However, steroids are steroid agents that temporarily alleviate symptoms, but they have the side effect of thinning the skin and ultimately worsening symptoms.
리기다 소나무(Pinus rigida Mill.)는 북아메리카 원산의 상록 교목으로서 1900년대부터 사방, 조림의 목적으로 수입되어 전국에 널리 재식되었다. 건조한 곳이나 습지에도 잘자라고 송충의 피해에도 강한 특징을 가진다. 리기다 소나무는 국내에서 원물의 수급이 용이하여 화장품 원료로서의 개발 가능성이 매우 크나 리기다 소나무 껍질의 생리활성에 대한 연구가 활발하지는 않았다.Rigida pine ( Pinus rigida Mill. ) is an evergreen tree native to North America and has been widely planted throughout the country since the 1900s, imported from all directions for afforestation purposes. It grows well in dry places or wetlands and is resistant to damage from caterpillars. Rigida pine has a high potential for development as a raw material for cosmetics because of its easy supply and demand in Korea, but research on the physiological activity of Rigida pine bark has not been active.
또한 천연물 소재는 빛이나 열에 대한 안정성이 낮고 피부 침투율이 낮아서 화장품 조성료로 많이 쓰이긴 하나 그 효능 효과를 담보하기는 어렵다. 천연물의 폴리페놀류나 다른 유효성분들은 지용성과 수용성 성분이 복합적으로 되어 있어 제형을 안정하게 하기가 힘든 단점이 있다. In addition, natural materials have low stability against light or heat and low skin penetration, so they are often used as cosmetic ingredients, but it is difficult to guarantee their efficacy. Natural polyphenols and other active ingredients have a disadvantage in that it is difficult to stabilize the formulation because the fat-soluble and water-soluble components are complex.
리포좀 전달체는 지용성과 수용성 성분들 동시에 봉입할 수 있어 천연물 물질을 봉입할 수 있는 가장 안정적인 전달체이다. 리포좀은 인지질 내부 코어에는 수용성물질을 봉입가능하고 지용성 물질은 인지질로 이루어진 전달체 막 사이에 끼어들어가는 형태로 물질을 봉입 가능하다.Liposome delivery system is the most stable delivery system capable of encapsulating natural substances because it can simultaneously encapsulate both fat-soluble and water-soluble components. Liposomes can encapsulate water-soluble substances in the inner core of phospholipids, and liposomes can encapsulate substances in a form in which the lipid-soluble substances are sandwiched between the membranes of a delivery system made of phospholipids.
본 발명자들은, 리기다 소나무 껍질 추출물을 에토좀에 봉입하여 화장료 조성물을 제조할 경우, 천연물 특유의 낮은 침투율을 극복하면서도, 효과적으로 피부염을 치료하고 피부를 진정시킬 수 있음을 개발하여 위와 같은 문제점을 해결하였다.The inventors of the present invention solved the above problems by developing that when a cosmetic composition is prepared by encapsulating Rigida pine bark extract in an ethosome, it can effectively treat dermatitis and soothe the skin while overcoming the low penetration rate peculiar to natural products. .
일 양상은 리기다 소나무 껍질 추출물(Pinus rigida bark extract) 또는 이의 분획물(Fraction)을 봉입하는 에토좀(Ethosome)을 포함하는 피부 진정, 피부 장벽 재생, 피부 간지러움, 피부 홍반 개선 또는 예방용 화장료 조성물을 제공한다.One aspect provides a cosmetic composition for skin soothing, skin barrier regeneration, skin itching, skin erythema improvement or prevention containing Ethosome encapsulating Pinus rigida bark extract or a fraction thereof do.
다른 양상은 리기다 소나무 껍질 추출물(Pinus rigida bark extract) 또는 이의 분획물(Fraction)을 봉입하는 에토좀(Ethosome)을 포함하는 피부염(Dermatitis) 개선 또는 예방용 화장료 조성물을 제공한다.Another aspect provides a cosmetic composition for improving or preventing dermatitis, including ethosome encapsulating Pinus rigida bark extract or a fraction thereof.
다른 양상은 리기다 소나무 껍질 추출물(Pinus rigida bark extract) 또는 이의 분획물(Fraction) 및 인지질을 교반하여 혼합액을 제조하는 단계; 및 상기 혼합액 및 정제수를 정제수와 교반하여 에토좀(Ethosome)을 제조하는 단계를 포함하는, 피부 진정, 피부 장벽 재생, 피부 간지러움, 피부 홍반 개선 또는 예방용 화장료 조성물의 제조 방법을 제공한다.Another aspect is preparing a mixed solution by stirring a pine bark extract (Pinus rigida bark extract) or a fraction thereof and a phospholipid; And it provides a method for preparing a cosmetic composition for skin soothing, skin barrier regeneration, skin itching, skin erythema improvement or prevention comprising the step of preparing ethosome by stirring the mixed solution and purified water with purified water.
일 양상은 리기다 소나무 껍질 추출물(Pinus rigida bark extract) 또는 이의 분획물(Fraction)을 봉입하는 에토좀(Ethosome)을 포함하는 피부 진정, 피부 장벽 재생, 피부 간지러움, 피부 홍반 개선 또는 예방용 화장료 조성물을 제공한다.One aspect provides a cosmetic composition for skin soothing, skin barrier regeneration, skin itching, skin erythema improvement or prevention containing Ethosome encapsulating Pinus rigida bark extract or a fraction thereof do.
다른 양상은 리기다 소나무 껍질 추출물(Pinus rigida bark extract) 또는 이의 분획물(Fraction)을 봉입하는 에토좀(Ethosome)을 포함하는 피부염(Dermatitis) 개선 또는 예방용 화장료 조성물을 제공한다.Another aspect provides a cosmetic composition for improving or preventing dermatitis, including ethosome encapsulating Pinus rigida bark extract or a fraction thereof.
본 명세서에서 용어 “리기다 소나무(Pinus rigida Bill.)”은 북아메리카 원산의 상록교목의 일종으로서, Pitch pine 이라고 명명되는 것일 수 있다.In this specification, the term "Pinus rigida Bill." is a kind of evergreen tree native to North America, and may be named Pitch pine.
본 명세서에서 용어, "추출물"이란, 리기다 소나무 껍질 또는 수피로부터 분리하여 얻은 물질을 의미한다. 바람직하게는, 외수피를 제거한 껍질에서부터 추출한 것일 수 있다.As used herein, the term "extract" refers to a material obtained by separating from the bark or bark of a pine tree Rigida. Preferably, it may be extracted from the skin from which the outer bark has been removed.
본 명세서에서 용어 “수피”는 수목의 형성층의 바깥쪽에 있는 모든 조직을 의미하며, 외수피 및 내수피로 구분되는 것일 수 있다.In this specification, the term “bark” refers to all tissues outside the cambium of a tree, and may be divided into outer bark and inner bark.
상기 리기다 소나무 껍질 추출물은 내수피 부분을 제거한, 외수피 부분 또는 최외곽 부분을 의미하는 것일 수 있다. The Rigida pine bark extract may refer to an outer bark part or an outermost part after removing the inner bark part.
상기 추출물의 용매는 정제수, 탄소수 1 내지 4의 알코올로부터 구성된 군으로부터 선택된 1종 이상을 포함하는 것일 수 있다. 바람직하게는, 상기 추출물의 용매는 알코올을 포함할 수 있다.The solvent of the extract may include one or more selected from the group consisting of purified water and alcohol having 1 to 4 carbon atoms. Preferably, the solvent of the extract may include alcohol.
일 구체예에 있어서, 상기 추출물의 추출 용매는 에탄올일 수 있다. 일 구체예에 있어서, 상기 에탄올은 50 내지 100 중량%, 55 내지 100 중량%, 60 내지 100 중량 %, 65 내지 100 중량%, 70 내지 100 중량%, 50 내지 95 중량%, 55 내지 95 중량%, 60 내지 95 중량%, 65 내지 95 중량%, 70 내지 95 중량%, 50 내지 90 중량%, 55 내지 90 중량%, 60 내지 90 중량%, 65 내지 90 중량%, 70 내지 90 중량%, 50 내지 85 중량%, 55 내지 85 중량%, 60 내지 85 중량%, 65 내지 85 중량%, 65 내지 85 중량% 또는 65 내지 80 중량%일 수 있다. 바람직하게는, 약 70% 알코올일 수 있다.In one embodiment, the extraction solvent of the extract may be ethanol. In one embodiment, the ethanol is 50 to 100% by weight, 55 to 100% by weight, 60 to 100% by weight, 65 to 100% by weight, 70 to 100% by weight, 50 to 95% by weight, 55 to 95% by weight , 60 to 95% by weight, 65 to 95% by weight, 70 to 95% by weight, 50 to 90% by weight, 55 to 90% by weight, 60 to 90% by weight, 65 to 90% by weight, 70 to 90% by weight, 50 to 85% by weight, 55 to 85% by weight, 60 to 85% by weight, 65 to 85% by weight, 65 to 85% by weight or 65 to 80% by weight. Preferably, it may be about 70% alcohol.
상기 추출물 또는 분획물은 건조 분말 형태를 포함하는 것일 수 있다. 상기 건조 분말 형태는, 냉각 콘덴서가 달린 감압 증류장치를 이용하여 증발되어 나오는 용매를 회수하면서 농축하고 건조시켜 수득한 것일 수 있다. The extract or fraction may be in the form of a dry powder. The dry powder form may be obtained by concentrating and drying while recovering the evaporated solvent using a vacuum distillation apparatus equipped with a cooling condenser.
상기 추출물 또는 분획물은, 리기다 소나무 추출 건조 분말 및 정제수를 혼합하여 제조한 것일 수 있다. The extract or fraction may be prepared by mixing Rigida pine extract dry powder and purified water.
일 구체예에 있어서, 상기 추출물 또는 분획물은, 상기 건조 분말을 0.01 중량% 내지 1 중량% 포함하는 것일 수 있다. 구체적으로, 상기 건조 분말은 0.02 내지 0.09, 0.03 내지 0.08 또는 0.03 내지 0.07 중량% 포함되는 것일 수 있다.In one embodiment, the extract or fraction may contain 0.01% to 1% by weight of the dry powder. Specifically, the dry powder may be contained in an amount of 0.02 to 0.09, 0.03 to 0.08 or 0.03 to 0.07% by weight.
일 구체예에 있어서, 상기 추출물 또는 분획물은, 정제수를 5 내지 20 중량% 포함하는 것일 수 있다. 구체적으로, 상기 정제수는 5 내지 18, 5 내지 15, 8 내지 15 또는 8 내지 13 중량% 포함하는 것일 수 있다.In one embodiment, the extract or fraction may contain 5 to 20% by weight of purified water. Specifically, the purified water may include 5 to 18, 5 to 15, 8 to 15, or 8 to 13% by weight.
상기 화장료 조성물은, 건조 분말 및 정제수를 혼합하여 추출물을 제조함으로써, 효율적인 운반 및 제조가 가능한 장점이 있는 것일 수 있다.The cosmetic composition may be advantageous in that it can be efficiently transported and prepared by preparing an extract by mixing dry powder and purified water.
상기 리기다 소나무 껍질 추출물 또는 이의 분획물은 카테킨(catehcin) 또는 퀘르시트린(quercitrin)을 포함하고 있는 것일 수 있다.The Rigida pine bark extract or a fraction thereof may contain catechin or quercitrin.
본 명세서에서 용어 “에토좀(Ethosome)”은 인지질을 에탄올에 혼합하여 제조한 지질구조체를 의미한다. 상기 에토좀은, 지질, 에탄올간의 상호작용으로 인하여, 특히 일부 지질층을 포함하는 피부에 대한 생리활성물질 투과에 효율적인 것일 수 있다.As used herein, the term "ethosome" refers to a lipid structure prepared by mixing phospholipids with ethanol. Due to the interaction between lipid and ethanol, the ethosome may be effective in permeating physiologically active substances through the skin including a part of the lipid layer.
상기 에토좀은 인지질 층을 포함하는 것일 수 있다. 또한, 상기 에토좀은 내부에 리기다 소나무 추출물 또는 이의 분획물이 봉입되어 추출물 또는 이의 분획물의 전달 안정성을 높이고, 전달률을 높이는 것일 수 있다.The ethosome may include a phospholipid layer. In addition, the ethosome may be encapsulated with a pine extract or a fraction thereof to increase delivery stability and a delivery rate of the extract or a fraction thereof.
상기 인지질 층은 이중층을 구성하며, 천연 인지질 또는 합성 지질을 포함할 수 있다. 상기 천연 인지질은 콜레스테롤, 난황 레시틴(포스파티딜콜린), 하이드로제네이티드레시틴, 대두 레시틴, 리조레시틴(Lysolecithin), 스핑고마이엘린(Sphingomyelin), 포스파티딜이노시톨, 포스파티드산, 포스파티딜에탄올아민, 포스파티딜세린, 포스파티딜글리세롤, 포스파티딜이노시톨-4,5-이인산, 카르디오리핀(Cardiolipin), 및 플라즈마로겐으로 구성되는 군으로부터 선택된 하나 이상의 것일 수 있다. 또한, 상기 합성 인지질은 디세틸포스페이트, 디스테아로일포스파티딜콜린, 디올레오일포스파티딜에탄올아민, 디팔미토일포스파티딜콜린, 디팔미토일포스파티딜에탄올아민, 디팔미토일포스파티딜세린, 엘레오스테아로일포스파티딜콜린, 엘레오스테아로일포스파티딜에탄올아민 및 엘레오스테아로일포스파티딜세린으로 구성된 군으로부터 선택된 하나 이상의 것일 수 있다. 바람직하게는, 하이드로제네이티드레시틴일 수 있다.The phospholipid layer constitutes a bilayer and may include natural phospholipids or synthetic lipids. The natural phospholipids are cholesterol, egg yolk lecithin (phosphatidylcholine), hydrogenated lecithin, soybean lecithin, lysolecithin, sphingomyelin, phosphatidylinositol, phosphatidic acid, phosphatidylethanolamine, phosphatidylserine, phosphatidylglycerol , phosphatidylinositol-4,5-diphosphate, cardiolipin, and plasma may be at least one selected from the group consisting of. In addition, the synthetic phospholipids include dicetyl phosphate, distearoylphosphatidylcholine, dioleoylphosphatidylethanolamine, dipalmitoylphosphatidylcholine, dipalmitoylphosphatidylethanolamine, dipalmitoylphosphatidylserine, eleostearoylphosphatidylcholine, eleostearo It may be at least one selected from the group consisting of ylphosphatidylethanolamine and eleostearoylphosphatidylserine. Preferably, it may be hydrogenated lecithin.
상기 에토좀은 에토좀 총 중량 대비 2.0 중량% 내지 5.0 중량%의 인지질을 포함하는 것일 수 있다. 구체적으로, 상기 인지질은 2.0 중량% 내지 4.5 중량%, 2.0 중량% 내지 4.0 중량%, 2.0 중량 내지 3.5 중량%, 2.5 내지 4.5 중량%, 2.5 중량% 내지 4.0 중량% 또는 2.5 중량 내지 3.5 중량%일 수 있다.The ethosomes may contain phospholipids in an amount of 2.0% to 5.0% by weight based on the total weight of the ethosomes. Specifically, the phospholipid is 2.0 wt% to 4.5 wt%, 2.0 wt% to 4.0 wt%, 2.0 wt% to 3.5 wt%, 2.5 to 4.5 wt%, 2.5 wt% to 4.0 wt%, or 2.5 wt% to 3.5 wt%. can
상기 에토좀은 에탄올을 포함하는 것일 수 있다. 상기 에토좀은 에토좀 총 중량 대비 에탄올을 1 중량% 내지 20 중량% 포함하는 것일 수 있다. 구체적으로, 상기 에토좀은 1 내지 18, 1 내지 15, 5 내지 15 또는 8 내지 13 중량%의 에탄올을 포함하는 것일 수 있다.The ethosome may contain ethanol. The ethosome may contain 1 wt% to 20 wt% of ethanol based on the total weight of the ethosome. Specifically, the ethosome may contain 1 to 18, 1 to 15, 5 to 15, or 8 to 13% by weight of ethanol.
상기 에토좀의 봉입률은 90% 이상 내지 100% 미만인 것일 수 있다. 구체적으로, 봉입률은 90 내지 99, 90 내지 98 또는 90 내지 97%인 것일 수 있다. The encapsulation rate of the ethosome may be 90% or more to less than 100%. Specifically, the encapsulation rate may be 90 to 99, 90 to 98 or 90 to 97%.
상기 에토좀은 조성물 총 중량에 대하여 0.001 중량% 내지 95 중량%, 예를 들면, 0.01 중량% 내지 80 중량%, 0.01 중량% 내지60 중량%, 0.01 중량% 내지40 중량%, 0.01 중량% 내지30 중량%, 0.01 중량% 내지20 중량%, 0.01 중량% 내지10 중량%, 0.01 중량% 내지5 중량%, 0.05 중량% 내지60 중량%, 0.05 중량% 내지40 중량%, 0.05 중량% 내지30 중량%, 0.05 중량% 내지20 중량%, 0.05 중량% 내지10 중량%, 0.05 중량% 내지5 중량%, 0.1 중량% 내지60 중량%, 0.1 중량% 내지40 중량%, 0.1 중량% 내지30 중량%, 0.1 중량% 내지20 중량%, 0.1 중량% 내지10 중량%, 또는0.1 중량% 내지5 중량%로 포함될 수 있다.The ethosome is 0.001% to 95% by weight, for example, 0.01% to 80% by weight, 0.01% to 60% by weight, 0.01% to 40% by weight, 0.01% to 30% by weight, based on the total weight of the composition. % by weight, 0.01% to 20% by weight, 0.01% to 10% by weight, 0.01% to 5% by weight, 0.05% to 60% by weight, 0.05% to 40% by weight, 0.05% to 30% by weight , 0.05 wt% to 20 wt%, 0.05 wt% to 10 wt%, 0.05 wt% to 5 wt%, 0.1 wt% to 60 wt%, 0.1 wt% to 40 wt%, 0.1 wt% to 30 wt%, 0.1 % to 20% by weight, 0.1% to 10% by weight, or 0.1% to 5% by weight.
일 구체예에 있어서, 상기 화장료 조성물은 피부 진정, 피부 장벽 재생, 피부 간지러움, 피부 트러블의 개선, 또는 피부 홍반을 개선 또는 예방하는 것일 수 있다. In one embodiment, the cosmetic composition may be to soothe the skin, regenerate the skin barrier, improve skin itching, improve skin troubles, or improve or prevent skin erythema.
일 구체예에 있어서, 상기 화장료 조성물은 피부염(Dermatitis) 증상을 개선 또는 예방하는 것일 수 있다. 상기 피부염의 증상은 피부 홍조, 피부 간지러움, 피부 손상, 피부 트러블, 피부 건조 및 피부 장벽 손상으로 이루어진 군으로부터 하나 이상을 포함하는 것일 수 있다.In one embodiment, the cosmetic composition may improve or prevent symptoms of dermatitis (dermatitis). Symptoms of the dermatitis may include one or more from the group consisting of skin flushing, skin itching, skin damage, skin trouble, skin dryness, and skin barrier damage.
본 명세서에서 용어 “피부 장벽 강화”는 피부의 가장 외각에 위치하는 각질층의 장벽 기능을 강화하여 피부 장벽 손상을 치료 및 개선하는 것을 의미한다.As used herein, the term "strengthen the skin barrier" means to treat and improve skin barrier damage by strengthening the barrier function of the stratum corneum located at the outermost layer of the skin.
일 구체예에 있어서, 상기 피부염은 아토피 피부염, 지루성 피부염, 건성 피부염, 접촉성 피부염, 화폐상 피부염 또는 알레르기성 접촉 피부염일 수 있다.In one embodiment, the dermatitis may be atopic dermatitis, seborrheic dermatitis, dry dermatitis, contact dermatitis, nummular dermatitis or allergic contact dermatitis.
본 명세서에서 용어, “아토피 피부염(atopic dermatitis)”은, “아토피”, “아토피성 질환” 등과 혼용될 수 있고, 피부에 발생하는 만성 알레르기 염증성 질환으로, 피부 건조, 각질이 많은 비늘피부, 피부 염증, 증가된 피부 투과성, 피부 표면 감염에 대한 취약성, 발작 및 가려움증 등의 증상을 수반하는 질환을 의미할 수 있다.As used herein, the term "atopic dermatitis" can be used interchangeably with "atopic", "atopic disease", etc., and is a chronic allergic inflammatory disease occurring on the skin, which is characterized by dry skin, scaly skin with a lot of horny skin, skin It can refer to a disease accompanied by symptoms such as inflammation, increased skin permeability, susceptibility to skin surface infection, seizures and itching.
일 구체예에 있어서, 상기 화장료 조성물은 경피수분손실량 또는 피부 홍반도를 감소시킴으로써, 피부 장벽을 강화하고 피부 홍반을 가라앉히며, 가려움증을 개선하는 효과가 있는 것일 수 있다.In one embodiment, the cosmetic composition may have the effect of strengthening the skin barrier, reducing skin erythema, and improving itching by reducing transepidermal water loss or skin erythema.
일 구체예에 있어서, 상기 화장료 조성물은, 항염증 화장료 조성물인 것일 수 있다. 상기 화장료 조성물은, 산화질소 또는 염증 사이토카인을 감소시키거나, 염증매개인자 단백질의 발현을 감소시킴으로써, 피부염을 효과적으로 예방 또는 개선하는 효과가 있을 수 있다.In one embodiment, the cosmetic composition may be an anti-inflammatory cosmetic composition. The cosmetic composition may have an effect of effectively preventing or improving dermatitis by reducing nitric oxide or inflammatory cytokines or reducing the expression of inflammatory mediator proteins.
일 구체예에 있어서, 상기 화장료 조성물은 상기 유효성분 외에 당업계에서 통상적으로 사용되는 첨가물 또는 부가물등을 더 첨가하여 스킨, 스킨토너, 부스터, 로션, 영양로션, 밀크로션, 마이스처로션, 영양크림, 맛사지 크림, 핸드크림, 에센스, 세럼, 팩, 마스크, 비누, 클렌징 폼, 콜렌징 로션, 클렌징 크림, 바디 클렌져, 바디로션, 샴푸, 린스, 메이크업베이스등 기초 및 색조화장품 제형을 갖는 것일 수 있다.In one embodiment, the cosmetic composition is a skin, skin toner, booster, lotion, nutrient lotion, milk lotion, misture lotion, and nutrient cream by adding additives or additives commonly used in the art in addition to the active ingredient. , Massage cream, hand cream, essence, serum, pack, mask, soap, cleansing foam, cleansing lotion, cleansing cream, body cleanser, body lotion, shampoo, conditioner, makeup base, etc. It may have basic and color cosmetic formulations. .
일 구체예에 있어서, 상기 화장료 조성물은 피부 외용제로 제형화될 수 있다. 구체적으로 상기 조성물은 액제, 연고제, 크림제, 로션제, 스프레이제, 패취제, 겔제, 또는 에어로졸제 제형일 수 있다.In one embodiment, the cosmetic composition may be formulated as an external skin preparation. Specifically, the composition may be a liquid, ointment, cream, lotion, spray, patch, gel, or aerosol formulation.
상기 화장료 조성물은 통상 화장품이나 의약품 등의 피부외용제에 사용되는 성분, 예를 들면 수성성분, 유성성분, 분말성분, 알코올류, 보습제, 증점제, 자외선흡수제, 미백제, 방부제, 산화방지제, 계면활성제, 향료, 색제, 각종 피부 영양제등을 필요에 따라서 적절하게 배합할 수 있다.The cosmetic composition is a component commonly used in external skin preparations such as cosmetics or pharmaceuticals, for example, water-based components, oil-based components, powder components, alcohols, moisturizers, thickeners, ultraviolet absorbers, whitening agents, preservatives, antioxidants, surfactants, and fragrances. , colorants, various skin nutrients, etc. can be appropriately blended as needed.
상기 화장료 조성물은, 에데트산이나트륨, 에데트산삼나트륨, 시트르산나트륨, 폴리인산나트륨, 메타인산나트륨, 글루콘산 등의 금속봉쇄제, 카페인, 탄닌, 벨라파밀, 감초추출물, 글라블리딘, 칼린의 과실의 열수추출물, 각종생약, 아세트산토코페롤, 글리틸리틴산, 트라넥삼산 및 그 유도체 또는 그 염등의 약제, 비타민C, 아스코르브산인산마그네슘, 아스코르브산글루코시드, 알부틴, 코지산, 글루코스, 프룩토스, 트레할로스 등의 당류 등도 적절하게 배합할 수 있다.The cosmetic composition contains metal sequestering agents such as disodium edetate, trisodium edetate, sodium citrate, sodium polyphosphate, sodium metaphosphate, and gluconic acid, caffeine, tannin, bellapamil, licorice extract, glabridin, and carlin. Hot-water extracts of fruits, various herbal medicines, tocopherol acetate, glycyrrhizic acid, tranexamic acid and its derivatives, or drugs such as its salts, vitamin C, magnesium ascorbic acid phosphate, ascorbic acid glucoside, arbutin, kojic acid, glucose, fructose, Sugars, such as trehalose, etc. can also be mix|blended suitably.
상기 화장료 조성물은 화장품학적으로 허용가능한 희석제 또는 담체를 추가적으로 포함할 수 있다. 상기 희석제는 유당, 옥수수 전분, 대두유, 미정질 셀룰로오스, 또는 만니톨, 활택제로는 스테아린산 마그네슘, 탈크, 또는 그 조합일 수 있다. 상기 담체는 부형제, 붕해제, 결합제, 활택제, 또는 그 조합일 수 있다. 상기 부형제는 미결정 셀룰로오즈, 유당, 저치환도 히드록시셀룰로오즈, 또는 그 조합일 수 있다. 상기 붕해제는 카르복시메틸셀룰로오스 칼슘, 전분글리콜산 나트륨, 무수인산일수소 칼슘, 또는 그 조합일 수 있다. 상기 결합제는 폴리비닐피롤리돈, 저치환도 히드록시프로필셀룰로오즈, 히드록시프로필셀룰로오즈, 또는 그 조합일 수 있다. 상기 활택제는 스테아린산 마그네슘, 이산화규소, 탈크, 또는 그 조합일 수 있다.The cosmetic composition may additionally include a cosmetically acceptable diluent or carrier. The diluent may be lactose, corn starch, soybean oil, microcrystalline cellulose, or mannitol, and magnesium stearate, talc, or a combination thereof as a lubricant. The carrier may be an excipient, a disintegrant, a binder, a lubricant, or a combination thereof. The excipient may be microcrystalline cellulose, lactose, low-substituted hydroxycellulose, or a combination thereof. The disintegrant may be calcium carboxymethylcellulose, sodium starch glycolate, calcium monohydrogen phosphate, or a combination thereof. The binder may be polyvinylpyrrolidone, low-substituted hydroxypropylcellulose, hydroxypropylcellulose, or a combination thereof. The lubricant may be magnesium stearate, silicon dioxide, talc, or a combination thereof.
다른 양상은 리기다 소나무 껍질 추출물(Pinus rigida Mill. bark extract) 또는 이의 분획물(Fraction) 및 인지질을 교반하여 혼합액을 제조하는 단계; 및 상기 혼합액 및 정제수를 교반하여 에토좀(Ethosome)을 제조하는 단계를 포함하는, 피부 진정, 피부 장벽 재생, 피부 간지러움, 피부 홍반 개선 또는 예방용 화장료 조성물을 제공한다.Another aspect is preparing a mixture by stirring a pine bark extract (Pinus rigida Mill. bark extract) or a fraction thereof (Fraction) and a phospholipid; And it provides a cosmetic composition for skin soothing, skin barrier regeneration, skin itching, skin erythema improvement or prevention comprising the step of preparing ethosome by stirring the mixed solution and purified water.
다른 양상은 리기다 소나무 껍질 추출물(Pinus rigida Mill. bark extract) 또는 이의 분획물(Fraction) 및 인지질을 교반하여 혼합액을 제조하는 단계; 및 상기 혼합액 정제수와 교반하여 에토좀(Ethosome)을 제조하는 단계를 포함하는, 피부염 개선 또는 예방용 화장료 조성물의 제조 방법을 제공한다.Another aspect is preparing a mixture by stirring a pine bark extract (Pinus rigida Mill. bark extract) or a fraction thereof (Fraction) and a phospholipid; And it provides a method for producing a cosmetic composition for improving or preventing dermatitis, comprising the step of preparing an ethosome by stirring with the mixed solution and purified water.
리기다 소나무 껍질, 추출물, 에토좀, 피부염, 화장료 조성물은 전술한 바와 같다.Rigida pine bark, extract, ethosome, dermatitis, and cosmetic composition are as described above.
상기 방법은, 리기다 추출물 또는 분획물의 건조 분말 및 정제수를 혼합하여 추출물 또는 분획물 용액을 제조하는 단계를 포함하는 것일 수 있다.The method may include preparing an extract or fraction solution by mixing dry powder of the Rigida extract or fraction and purified water.
상기 방법은, 가온조건에서 인지질 및 에탄올을 혼합하여, 인지질 용액을 제조하는 단계를 포함하는 것일 수 있다.The method may include preparing a phospholipid solution by mixing phospholipid and ethanol under warm conditions.
상기 인지질은 에토좀 총 중량 대비 2.0 중량% 내지 5.0 중량% 포함되는 것일 수 있다. 구체적으로, 상기 인지질은 2.0 중량% 내지 4.5 중량%, 2.0 중량% 내지 4.0 중량%, 2.0 중량 내지 3.5 중량%, 2.5 내지 4.5 중량%, 2.5 중량% 내지 4.0 중량% 또는 2.5 중량 내지 3.5 중량%일 수 있다.The phospholipid may be included in an amount of 2.0% to 5.0% by weight based on the total weight of the ethosome. Specifically, the phospholipid is 2.0 wt% to 4.5 wt%, 2.0 wt% to 4.0 wt%, 2.0 wt% to 3.5 wt%, 2.5 to 4.5 wt%, 2.5 wt% to 4.0 wt%, or 2.5 wt% to 3.5 wt%. can
상기 에탄올은 포함하는 것일 수 있다. 상기 에탄올은 에토좀 총 중량 대비 1 중량% 내지 20 중량% 포함되는 것일 수 있다. 구체적으로, 상기 에탄올은 1 내지 18, 1 내지 15, 5 내지 15 또는 8 내지 13 중량% 포함되는 것일 수 있다.The ethanol may contain. The ethanol may be contained in an amount of 1% to 20% by weight based on the total weight of the ethosome. Specifically, the ethanol may be contained in an amount of 1 to 18, 1 to 15, 5 to 15, or 8 to 13% by weight.
상기 방법은, 상기 추출물 또는 이의 분획물 용액 및 인지질 용액을 가온조건에서 혼합하여 에토좀 용액을 제조하는 단계; 및 상기 용액을 과량의 정제수와 교반하여 분산 및 균질화하는 단계를 포함하는 것일 수 있다.The method may include preparing an ethosome solution by mixing the extract or a solution of a fraction thereof and a phospholipid solution under warm conditions; and dispersing and homogenizing the solution by stirring with an excess of purified water.
상기 방법은, 교반된 용액을 냉각하는 단계를 추가적으로 포함하는 것일 수 있다.The method may further include cooling the stirred solution.
상기 방법은, 상기 에토좀을 포함하는 화장료 조성물을 제조하는 단계를 추가적으로 포함하는 것일 수 있다.The method may further include preparing a cosmetic composition containing the ethosome.
상기 에토좀은 화장료 조성물 총 중량에 대하여 0.001 중량% 내지 95 중량%, 예를 들면, 0.01 중량% 내지 80 중량%, 0.01 중량% 내지60 중량%, 0.01 중량% 내지40 중량%, 0.01 중량% 내지30 중량%, 0.01 중량% 내지20 중량%, 0.01 중량% 내지10 중량%, 0.01 중량% 내지5 중량%, 0.05 중량% 내지60 중량%, 0.05 중량% 내지40 중량%, 0.05 중량% 내지30 중량%, 0.05 중량% 내지20 중량%, 0.05 중량% 내지10 중량%, 0.05 중량% 내지5 중량%, 0.1 중량% 내지60 중량%, 0.1 중량% 내지40 중량%, 0.1 중량% 내지30 중량%, 0.1 중량% 내지20 중량%, 0.1 중량% 내지10 중량%, 또는0.1 중량% 내지5 중량%로 포함될 수 있다.The ethosome is 0.001% to 95% by weight, for example, 0.01% to 80% by weight, 0.01% to 60% by weight, 0.01% to 40% by weight, 0.01% to 40% by weight, based on the total weight of the cosmetic composition 30 wt%, 0.01 wt% to 20 wt%, 0.01 wt% to 10 wt%, 0.01 wt% to 5 wt%, 0.05 wt% to 60 wt%, 0.05 wt% to 40 wt%, 0.05 wt% to 30 wt% %, 0.05% to 20%, 0.05% to 10%, 0.05% to 5%, 0.1% to 60%, 0.1% to 40%, 0.1% to 30%, 0.1% to 20% by weight, 0.1% to 10% by weight, or 0.1% to 5% by weight.
일 양상의 조성물은 리기다 소나무 껍질 추출물을 나노 에토좀에 봉입함으로서, 리기다 소나무 껍질 추출물의 낮은 침투율을 극복하면서도, 효과적으로 피부염을 치료하고 피부를 진정시키며, 피부 홍반도를 낮추고 피부 보습이 우수한 효과가 있다.One aspect of the composition encapsulates the Rigida pine bark extract in nano ethosomes, thereby overcoming the low penetration rate of the Rigida pine bark extract, effectively treating dermatitis, soothing the skin, reducing skin erythema, and having excellent skin moisturizing effects. .
도 1은 리기다 소나무 껍질 추출물을 봉입한 나노 에토좀의 입자 크기를 측정한 그래프이다.
도 2은 리기다 소나무 껍질 추출물 봉입 나노 에토좀의 세포독성을 MTT assay로 평가한 도이다.
도 3는 리기다 소나무 껍질 추출물 및 상기 추출물이 봉입된 나노 에토좀의 Nitric oxide(NO) 생성량을 측정한 그래프이다.
도 4는 리가다 소나무 껍질 추출물 및 상기 추출물이 봉입된 나노 에토좀의 사이토카인(Cytokine) 생성량을 나타낸 그래프이다.
도 5는 리가다 소나무 껍질 추출물 및 상기 추출물이 봉입된 나노 에토좀의 TNF-α 생성량을 나타낸 그래프이다.
도 6은 리가다 소나무 껍질 추출물 및 상기 추출물이 봉입된 나노 에토좀의 IL-6 생성량을 나타낸 그래프이다.
도 7은 리가다 소나무 껍질 추출물 및 상기 추출물이 봉입된 나노 에토좀의 웨스턴 블랏 결과를 나타낸 도이다.
도 8은 프라츠 분산 세포(Franz Diffusion Cell) 실험을 통해 리가다 소나무 껍질 추출물 및 상기 추출물이 봉입된 나노 에토좀의 피부 흡수율을 나타낸 그래프이다.
도 9는 리기다 소나무 껍질 추출물을 봉입한 나노 에토좀이 포함된 크림의 피부 진정 효과를 나타낸 이미지이다.1 is a graph measuring the particle size of nano ethosomes encapsulated with Rigida pine bark extract.
Figure 2 is a diagram evaluating the cytotoxicity of Rigida pine bark extract encapsulated nano ethosomes by MTT assay.
Figure 3 is a graph measuring the amount of nitric oxide (NO) produced by Rigida pine bark extract and nano ethosomes encapsulated with the extract.
Figure 4 is a graph showing the amount of cytokine (Cytokine) produced by the ligada pine bark extract and nano ethosomes encapsulated with the extract.
Figure 5 is a graph showing the amount of TNF-α production of Rigada pine bark extract and nano ethosomes encapsulated with the extract.
6 is a graph showing the amount of IL-6 produced by Rigada pine bark extract and nano-ethosomes encapsulated with the extract.
Figure 7 is a diagram showing the Western blot results of Rigada pine bark extract and nano-ethosomes encapsulated with the extract.
8 is a graph showing the skin absorption rate of Rigada pine bark extract and nano ethosomes encapsulated with the extract through a Franz Diffusion Cell experiment.
9 is an image showing the skin soothing effect of a cream containing nano-ethosomes encapsulated with Rigida pine bark extract.
이하 본 발명을 실시예를 통하여 보다 상세하게 설명한다. 그러나, 이들 실시예는 하나 이상의 구체예를 예시적으로 설명하기 위한 것으로 본 발명의 범위가 이들 실시예에 한정되는 것은 아니다.Hereinafter, the present invention will be described in more detail through examples. However, these examples are intended to illustrate one or more specific examples, and the scope of the present invention is not limited to these examples.
실시예 1: 리기다 소나무 껍질 추출물을 봉입한 나노 에토좀 제조Example 1: Preparation of nano ethosomes encapsulated with Rigida pine bark extract
1.1 1.1 리기다 소나무 껍질 추출물의 제조Preparation of Rigida Pine Bark Extract
리기다 소나무 껍질 추출물을 제조하기 위하여, 리기다 소나무(Pinus rigida Mill.)의 껍질 부분에서 수피 부분을 제거한 후, 2 내지 3cm의 크기로 세절하였다. 다음으로, 상기 손질한 리기다 소나무 껍질을 정제수로 세척한 후 서늘한 곳에서 건조한 후, 원물 중량의 9배의 70% 에탄올을 추가하여, 24시간 침지하여 추출하였다. 상기 추출액을 여과지(Filter paper, Whatman NO.2 Tokyo, Japan)를 사용하여 여과하였다. 여과 후, 추출 및 여과 과정을 총 3회 반복함으로써, 추출물을 완성하였다.In order to prepare the bark extract of Rigida pine, after removing the bark from the bark of Rigida pine (Pinus rigida Mill.), it was cut into pieces of 2 to 3 cm in size. Next, the trimmed Rigida pine bark was washed with purified water, dried in a cool place, and 70% ethanol nine times the weight of the raw material was added, followed by soaking for 24 hours for extraction. The extract was filtered using filter paper (Whatman NO.2 Tokyo, Japan). After filtration, the extract was completed by repeating the extraction and filtration processes a total of three times.
상기 추출물을 진공 감압 농축기를 사용하여 40℃, 40hPa 조건에서 감압 농축하여 에탄올을 모두 제거한 후 농축액을 회수하였다. 다음으로, 상기 농축액을 -80℃에서 동결한 후 동결건조기를 통해 건조된 분말 형태로 제조하였다.The extract was concentrated under reduced pressure at 40° C. and 40 hPa using a vacuum concentrator to remove all ethanol, and then the concentrate was recovered. Next, the concentrate was frozen at -80 ° C and then dried through a lyophilizer to prepare a powder form.
1.2 리기다 소나무 껍질 추출물을 봉입한 나노 에토좀 제조1.2 Preparation of nano-ethosomes encapsulated with Rigida pine bark extract
상기 제조예 1의 추출물 분말을 정제수에 첨가하여 용해시켜 코어 용액을 제조하였다. 용해 시, 리기다 소나무 껍질 추출물이 수용성 또는 지용성 성분을 포함하고 있기 문에, 가온조건으로 교반하여 용해하였다. 이 때, 리기다 소나무 껍질 추출물 분말은 총 나노 에토좀 중량 대비 0.05중량%, 정제수는 10.0중량% 포함되었다.A core solution was prepared by adding and dissolving the extract powder of Preparation Example 1 in purified water. When dissolving, since the Rigida pine bark extract contains water-soluble or fat-soluble components, it was dissolved by stirring under warm conditions. At this time, 0.05% by weight of Rigida pine bark extract powder and 10.0% by weight of purified water were included based on the total weight of nano-ethosomes.
인지질 용해액을 제조하기 위해, 이중막을 구성하는 인지질을 에탄올에 첨가하여 가온조건에서 교반하여 용해시켰다. 이 때 인지질은 하이드로제네이티드레시틴을 사용하였으며, 상기 하이드로제네이티드레시틴은 총 나노 에토좀 중량 대비 3.0중량% 포함되었다.To prepare a phospholipid solution, phospholipids constituting the double membrane were added to ethanol and dissolved by stirring under warm conditions. At this time, hydrogenated lecithin was used as the phospholipid, and the hydrogenated lecithin was included in an amount of 3.0% by weight based on the total weight of nano-ethosomes.
다음으로, 상기 추출물 용액 및 상기 인지질 용해액을 혼합하여 10분이상 가온조건에서 교반함으로써, 추출물을 인지질 용해액에 수화시켜 수화액정상 용액을 제조하였다. Next, by mixing the extract solution and the phospholipid solution and stirring under a heating condition for 10 minutes or more, the extract was hydrated in the phospholipid solution to prepare a hydrated liquid crystal solution.
상기 수화액정상 용액에 정제수를 과량 첨가하여 가온조건에서 30분간 교반하여 분산 및 균질화를 진행하고, 이후 상온으로 서서히 냉각하며 추가 교반을 진행하였다.An excessive amount of purified water was added to the hydrated liquid crystal solution, and the mixture was stirred for 30 minutes under a warming condition to disperse and homogenize, and then slowly cooled to room temperature and further stirred.
교반이 완료된 나노 에토좀 용액은 호모믹서를 통해 3000RPM 이상으로 10분간 균질화한 후 마이크로플루다이져로 1000bar, 3회 통과하여 추가적인 균질화를 진행함으로써, 리기나 소나무 껍질 추출물을 봉입한 나노 에토좀 용액을 제조하였다.The stirred nano-ethosome solution was homogenized at 3000 RPM or more for 10 minutes through a homomixer, and then passed through a microfluidizer at 1000 bar three times to further homogenize, thereby obtaining a nano-ethosome solution encapsulated with Rigi or pine bark extract. manufactured.
실시예 2: 인지질 첨가량을 2.0중량%로 한 나노 에토좀 제조Example 2: Preparation of nano-ethosomes with 2.0% by weight of phospholipid added
실시예 2은 실시예 1에서 이중막을 구성하는 하이드로제네이티드레시틴의 함량을 2.0중량%로 변경한 것을 제외하고는 동일하게 제조하였다.Example 2 was prepared in the same manner as in Example 1 except that the content of hydrogenated lecithin constituting the double membrane was changed to 2.0% by weight.
실시예 3: 인지질 첨가량을 1.0중량%로 한 나노 에토좀 제조Example 3: Preparation of nano-ethosomes with 1.0% by weight of phospholipid added
실시예 3는 실시예 1에서 이중막을 구성하는 하이드로제네이티드레시틴의 함량을 1.0중량%로 변경한 것을 제외하고는 동일하게 제조하였다.Example 3 was prepared in the same manner as in Example 1 except that the content of hydrogenated lecithin constituting the double membrane was changed to 1.0% by weight.
실시예 4: 리기다 소나무 껍질 추출물을 0.1중량%로 한 나노 에토좀 제조Example 4: Preparation of nano ethosomes with 0.1% by weight of Rigida pine bark extract
실시예 4는 실시예 1에서 리기다 소나무 껍질 추출물 분말을 0.1중량%로 변경한 것을 제외하고는 동일하게 제조하였다.Example 4 was prepared in the same manner as in Example 1, except that the Rigida pine bark extract powder was changed to 0.1% by weight.
실시예 5: 정제수와 에탄올을 5.0중량%로 한 나노 에토좀 제조Example 5: Preparation of Nano Ethosomes with Purified Water and Ethanol at 5.0% by Weight
실시예 5는 실시예 1에서 정제수와 에탄올을 5.0중량%로 변경한 것을 제외하고는 동일하게 제조하였다.Example 5 was prepared in the same manner as in Example 1 except for changing the purified water and ethanol to 5.0% by weight.
비교예 1Comparative Example 1
비교예 1는 실시예 1에서 나노 에토좀을 구성하는 성분을 모두 제외하고 리기다 소나무 껍질 추출물 분말만 정제수에 용해하여 제조하였다.Comparative Example 1 was prepared by dissolving only Rigida pine bark extract powder in purified water, excluding all components constituting the nano-ethosome in Example 1.
비교예 2Comparative Example 2
비교예 2는 실시예 1에서 리기다 소나무 껍질 추출물 분말을 첨가하지 않은 것을 제외하고는 동일하게 제조하였다Comparative Example 2 was prepared in the same manner as in Example 1, except that Rigida pine bark extract powder was not added.
표 1은 전술한 실시예 1 내지 실시예 5와 비교예 1 내지 비교예 2의 함량을 나타낸 것이다. Table 1 shows the contents of Examples 1 to 5 and Comparative Examples 1 to 2 described above.
[제조예][Production Example]
제조예 1. 나노 에토좀을 포함하는 화장료 조성물의 제조Preparation Example 1. Preparation of a cosmetic composition containing nano ethosomes
상기 실시예 2인 리기다 소나무 껍질 추출물 봉입 나노 에토좀이 함유되어 있는 크림 제형의 화장품 조성물을 하기 표 2의 함량과 같이 제조하였다. The cosmetic composition of the cream formulation of Example 2 containing Rigida pine bark extract encapsulated nano-ethosomes was prepared according to the contents of Table 2 below.
표 2는 제조예 1의 화장료 조성물의 함량을 나타낸 것이다.Table 2 shows the content of the cosmetic composition of Preparation Example 1.
[실험예][Experimental example]
실험예 1 : 리기다 소나무 껍질 추출물의 나노 에토좀 입자 크기 및 안정성 분석Experimental Example 1: Analysis of Nano Ethosome Particle Size and Stability of Rigida Pine Bark Extract
상기 실시예 및 비교예의나노 에토좀의 평균 입자 크기를 분석하기 위해 나노입자 분석기(Nanophox, Sympatec, 독일)로 입자 크기를 측정하였다. 또한, 나노 에토좀의 장기 보관 시 발생되는 응집, 침전, 변색 등의 불안정성을 확인하기 위한 장기 안정성 분석을 진행하였다.In order to analyze the average particle size of the nanoethosomes of the above Examples and Comparative Examples, the particle size was measured using a nanoparticle analyzer (Nanophox, Sympatec, Germany). In addition, long-term stability analysis was conducted to confirm instability such as aggregation, precipitation, discoloration, etc. that occurs during long-term storage of nano-ethosomes.
장기 안정성 분석은 실시예 및 비교예에서 제조된 나노 에토좀 용액을 45℃ 항온 챔버에 배치하여 1일, 10일, 20일, 30일간 경과시점을 기준으로 육안관찰을 통한 형태 변화를 관찰하여 진행하였다. Long-term stability analysis was carried out by placing the nano-ethosome solutions prepared in Examples and Comparative Examples in a constant temperature chamber at 45 ° C and observing morphological changes through visual observation based on the elapsed time of 1 day, 10 days, 20 days, and 30 days did
도 1은 리기다 소나무 껍질 추출물을 봉입한 나노 에토좀의 입자 크기를 측정한 그래프이다.1 is a graph measuring the particle size of nano ethosomes encapsulated with Rigida pine bark extract.
표 3은 나노에토좀의 평균 입자 크기 및 일자별 안정성에 대한 결과를 나타낸 것이다.Table 3 shows the results of the average particle size and daily stability of nanoetosomes.
(nm)average particle size
(nm)
그 결과, 도 1 및 표 3에 나타낸 것과 같이, 실시예 2 내지 실시예 6의 경우 300nm이하의 입자크기로 측정되었으며, 나노 에토좀을 구성하는 인지질의 구성함량이 증가할수록 입자크기가 커지고, 코어에 위치하는 리기다 소나무 껍질 추출물의 함량이 증가할수록 입자크기가 커지는 것을 확인하였다.As a result, as shown in Figure 1 and Table 3, in the case of Examples 2 to 6, the particle size was measured at 300 nm or less, and as the content of the phospholipid constituting the nano-ethosome increases, the particle size increases, and the core It was confirmed that the particle size increased as the content of the Rigida pine bark extract located in .
특히, 비교예 2의 경우 리기다 소나무 껍질 추출물이 봉입되지 않은 나노 에토좀으로서 약 100nm의 입자크기를 형성하고 있어, 리기다 소나무 껍질 추출물의 봉입으로 인해 전체적인 나노 에토좀의 입자크기가 커지는 것을 확인하였다.In particular, in the case of Comparative Example 2, it was confirmed that the Rigida pine bark extract formed a particle size of about 100 nm as nano ethosomes that were not encapsulated, and that the encapsulation of the Rigida pine bark extract increased the overall particle size of the nano ethosomes.
실험예 2 : 리기다 소나무 껍질 추출물의 나노 에토좀 봉입 효율 측정Experimental Example 2: Measurement of Encapsulation Efficiency of Nano Etosomes of Rigida Pine Bark Extract
상기 실시예 및 비교예로부터 제조된 나노 에토좀의 봉입효율을 측정하기 위해 고성능액체크로마토그래프(High Performance Chromatography, HPLC, Shimadzu, 일본)로 전체 함량과 미봉입된 함량 분석을 진행하였다. 이후, 분석 결과를 바탕으로 하기 식 1을 이용하여 봉입효율을 계산하였다.In order to measure the encapsulation efficiency of the nano-ethosomes prepared from the above Examples and Comparative Examples, analysis of the total content and the unencapsulated content was performed with High Performance Chromatography (HPLC, Shimadzu, Japan). Thereafter, the encapsulation efficiency was calculated using
[식 1][Equation 1]
Entrapment Efficiency(EE) = (WTotal-WFree)/WTotal x 100 Entrapment Efficiency (EE) = (W Total -W Free )/W Total x 100
상기 식 1에서 EE는 표준물질(리기다 소나무 껍질 추출물 중 카테킨)의 봉입효율을 의미하고, WTotal는 전체 표준물질 농도를 의미하며, WFree는 미봉입된 표준물질 농도를 의미한다. 아래 표 4는 실험 결과로 나타난 봉입효율을 수치로 계산한 것이다.In
효율 (%)enclosed
efficiency (%)
봉입효율 측정을 위한 지표물질은 Catechin과 Quercitrin으로 Quercitrin은 지용성 물질로 정제수에 매우 낮은 용해도를 가지고 있으며, Catechin은 정제수에 상대적으로 높은 용해도를 가지고 있지만, 수용성과 지용성 모두 봉입할 수 있는 나노 에토좀의 특성으로 인해 Catechin과 Quercitrin 모두 실시예 2 내지 실시예 4에서 90%이상의 높은 봉입효율을 나타내었다.Indicator substances for measuring encapsulation efficiency are Catechin and Quercitrin. Quercitrin is a fat-soluble substance and has very low solubility in purified water. Catechin has relatively high solubility in purified water. Due to the characteristics, both Catechin and Quercitrin showed high encapsulation efficiency of 90% or more in Examples 2 to 4.
실험예 3 : 리기다 소나무 껍질 추출물의 나노 에토좀 성분 안정성 실험Experimental Example 3: Stability test of nano ethosomal component of Rigida pine bark extract
상기 실시예 2 및 비교예 1에 함유되어 있는 성분 안정성 분석을 위해 각 시료를 45℃ 항온 챔버에 배치하여 1일, 5일, 10일, 15일, 20일, 25일, 30일간 경과시점을 기준으로 시료를 채취하여 고성능액체크로마토그래프를 통해 유효성분인 카테킨(Catechin)의 함량 분석을 진행하였으며, 각 결과는 하기 표 5에 나타내었다.For the stability analysis of the components contained in Example 2 and Comparative Example 1, each sample was placed in a 45 ° C. A sample was taken as a standard and the content of catechin, an active ingredient, was analyzed through a high-performance liquid chromatograph, and each result is shown in Table 5 below.
표 5는 각 경과시점의 고성능액체크로마토그래프 결과를 나타낸 것이다. 표 5의 단위는 μg/mL이다.Table 5 shows the results of high-performance liquid chromatography at each time point. The unit of Table 5 is μg/mL.
그 결과, 표 5에 나타낸 것과 같이, 실시예 2의 경우 30일 경과한 시점에서도 유효성분의 함량이 1일차 대비하여 16%만 감소하였지만, 비교예 1인 리기다 소나무 껍질 추출물 수용액의 경우 50%로 감소하여 실질적으로 나노에토좀의 유효성분이 감소하는 것을 확인하였다. As a result, as shown in Table 5, in the case of Example 2, even after 30 days, the content of the active ingredient decreased by only 16% compared to the first day, but in the case of the aqueous solution of Rigida pine bark extract of Comparative Example 1, it decreased to 50%. It was confirmed that the active ingredient of nanoetosomes was substantially reduced by decreasing.
실험예 4 : 리기다 소나무 껍질 추출물의 나노 에토좀 세포 독성 시험 결과Experimental Example 4: Results of nano-ethsome cytotoxicity test of Rigida pine bark extract
상기 실시예 2 및 비교예 1, 비교예 2의 세포 독성 측정을 위해 MTT assay를 진행하였다.MTT assay was performed to measure the cytotoxicity of Example 2 and Comparative Examples 1 and 2.
macrophage Raw 264.7 cell을 96 웰 플레이트에 4 x 104 cells/mL로 분주한 후, Fetal bovine serum free DMEM 배지로 리기나 소나무 껍질 추출물과 나노 에토좀 시료를 각각 6.25, 12.5, 25 ug/mL의 농도로 조제하여 20uL 첨가하였다. 다음으로, 배양 후 생존 세포에 MTT (5mg/mL)를 20uL씩 처치 후 4시간 뒤 배지를 제거하고 생성된 formazan crystal 99% ethanol과 DMSO를 1:1 비율로 혼합하여 200uL씩 처치한 뒤, 30분 후 540nm에서 흡광도를 측정하였다.After dispensing 4 x 104 cells/mL of macrophage Raw 264.7 cells in a 96-well plate, Rigina pine bark extract and nanoetosome samples were added to Fetal bovine serum free DMEM medium at concentrations of 6.25, 12.5, and 25 ug/mL, respectively. It was prepared and added 20uL. Next, after culturing, the viable cells were treated with 20uL of MTT (5mg/mL), and after 4 hours, the medium was removed, and the resulting formazan crystal 99% ethanol and DMSO were mixed in a 1:1 ratio and treated with 200uL each, followed by 30 Minutes later, absorbance was measured at 540 nm.
도 2은 리기다 소나무 껍질 추출물 봉입 나노 에토좀의 세포독성을 MTT assay로 평가한 도이다.Figure 2 is a diagram evaluating the cytotoxicity of Rigida pine bark extract encapsulated nano ethosomes by MTT assay.
그 결과, 도 2에서 나타낸 바와 같이, 25μg/mL까지 80%이상의 세포 생존율을 보였다. 하기에서는 보다 안정적인 세포 실험을 위해 12.5μg/mL 처리 농도로 세포 실험을 진행하였다.As a result, as shown in FIG. 2, the cell viability was 80% or more up to 25 μg/mL. In the following, cell experiments were conducted at a treatment concentration of 12.5 μg/mL for more stable cell experiments.
실험예 5 : 리기다 소나무 껍질 추출물 나노 에토좀의 NO 생성량 측정Experimental Example 5: Measurement of NO production of Rigida pine bark extract nano ethosomes
실시예 1 및 비교예 1 내지 비교예 2의 항염증 효과를 확인하기 위하여, 대표적인 염증매개인자(Inflammatory mediators)인산화질소(Nitric oxide: NO) 생성량 실험을 진행하였다.In order to confirm the anti-inflammatory effect of Example 1 and Comparative Examples 1 to 2, a representative inflammatory mediator, nitric oxide (NO) production test was conducted.
구체적으로, 웰 플레이트에 Raw 264.7 세포를 5 x 105 cells/mL로 분주하여 18 내지 24시간 배양한 뒤 Fetal bovine serum free DMEM 배지로 교체 후 시료를 농도별로 조제하였다. 이후, 조제한 시료를 300uL 첨가하여 배양한 다음, 18시간 후 96 웰 플레이트에 상층액과 그리스 리젠트(Griess regent)를 각각 100uL 처치 후, 15분 반응시켜 540nm에서 흡광도를 측정하였다.Specifically, Raw 264.7 cells were dispensed in a well plate at 5 x 105 cells/mL, cultured for 18 to 24 hours, and then replaced with Fetal bovine serum free DMEM medium, and samples were prepared by concentration. Thereafter, the prepared sample was cultured by adding 300uL, and after 18 hours, 100uL of the supernatant and Griess regent were treated in a 96-well plate, followed by reaction for 15 minutes to measure absorbance at 540nm.
도 3는 리기다 소나무 껍질 추출물 및 상기 추출물이 봉입된 나노 에토좀의 Nitric oxide(NO) 생성량을 측정한 그래프이다.Figure 3 is a graph measuring the amount of nitric oxide (NO) produced by Rigida pine bark extract and nano ethosomes encapsulated with the extract.
그 결과, 도 3에 나타난 바와 같이 실시예 2는 12.5μg/mL의 농도에서 77.7%의 NO 생성량을 보여 , 비교예 1은 72%, 비교예 2는 84.6%의 NO 생성량을 가짐을 확인하였다. 이러한 결과는, 실시예 2가 22.3%의 높은 NO 생성 저해율을 가짐으로써, 항염증 효과가 현저한 것을 의미한다.As a result, as shown in Figure 3, Example 2 showed a NO production amount of 77.7% at a concentration of 12.5 μg / mL, Comparative Example 1 was confirmed to have a NO production amount of 72%, Comparative Example 2 84.6%. These results mean that Example 2 has a high NO production inhibition rate of 22.3%, so that the anti-inflammatory effect is remarkable.
실험예 6 : 리기다 소나무 껍질 추출물 나노 에토좀의 Cytokine 생성량 측정Experimental Example 6: Measurement of Cytokine Production of Rigida Pine Bark Extract Nano Ethosomes
실시예 1 및 비교예 1 내지 비교예 2의 항염증 효과를 확인하기 위하여, tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), interleukin-6 (IL-6)가 inducible Nitric Oxide Synthasel (iNOS)와 cyclooxygenase-2 (COX-2)와 같은 염증성 사이토카인의 생성량을 측정하였다.In order to confirm the anti-inflammatory effect of Example 1 and Comparative Examples 1 to 2, tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), interleukin-6 (IL-6) were inducible Production of inflammatory cytokines such as Nitric Oxide Synthasel (iNOS) and cyclooxygenase-2 (COX-2) was measured.
구체적으로, 웰 플레이트에 Raw 264.7 세포를 5 x 105 cells/mL로 분주하여 24 시간동안 배양하고, LPS 1μg/mL와 시료 12.5μg/mL의 농도로 처리한 다음 18 시간동안 재배양하였다. 이후 세포 배양액을 취하여 ELISA kit(BD Biosciences, San Diego, CA, USA)를 사용하고 ELISA reader로 450nm에서 흡광도를 측정한 후 각 검량곡선을 바탕으로 염증성 사이토카인 IL-6, IL-1β, TNF-α의 생성량을 산출하였다.Specifically, Raw 264.7 cells were dispensed in a well plate at 5 x 105 cells/mL, cultured for 24 hours, treated with 1 μg/mL of LPS and 12.5 μg/mL of sample, and then cultured for 18 hours. Afterwards, the cell culture medium is taken, an ELISA kit (BD Biosciences, San Diego, CA, USA) is used, and the absorbance is measured at 450 nm with an ELISA reader. Based on each calibration curve, inflammatory cytokines IL-6, IL-1β, TNF- The production amount of α was calculated.
도 4는 리가다 소나무 껍질 추출물 및 상기 추출물이 봉입된 나노 에토좀의 사이토카인(Cytokine) 생성량을 나타낸 그래프이다. Figure 4 is a graph showing the amount of cytokine (Cytokine) produced by the ligada pine bark extract and nano ethosomes encapsulated with the extract.
도 5는 리가다 소나무 껍질 추출물 및 상기 추출물이 봉입된 나노 에토좀의 TNF-α 생성량을 나타낸 그래프이다. Figure 5 is a graph showing the amount of TNF-α production of Rigada pine bark extract and nano ethosomes encapsulated with the extract.
도 6은 리가다 소나무 껍질 추출물 및 상기 추출물이 봉입된 나노 에토좀의 IL-6 생성량을 나타낸 그래프이다.6 is a graph showing the amount of IL-6 produced by Rigada pine bark extract and nano-ethosomes encapsulated with the extract.
그 결과 도 4 내지 도 6에 나타난 바와 같이 IL-6는 실시예 2에서 23.9%의 저해율을 보였으며, IL-1β는 14.6%, TNF-α는 29.6%의 저해율을 보였다. 이러한 결과는, 일 양상의 조성물이 높은 사이토카인 생성 저해율을 가짐으로써, 항염증 효과가 현저한 것을 의미한다.As a result, as shown in FIGS. 4 to 6, IL-6 showed an inhibition rate of 23.9% in Example 2, IL-1β showed an inhibition rate of 14.6%, and TNF-α showed an inhibition rate of 29.6%. These results mean that the anti-inflammatory effect is remarkable because the composition of one aspect has a high cytokine production inhibition rate.
실험예 7 : Western blot을 통한 리기다 추출물 나노 에토좀의 iNOS, COX2 단백질 발현율 측정Experimental Example 7: Measurement of iNOS and COX2 protein expression in Rigida extract nanoetosomes through Western blot
실시예 1 및 비교예 1 내지 비교예 2의 항염증 효과를 확인하기 위하여, 웨스턴 블랏을 이용하여 각 조성물에 의한 염증매개인자 inducible nitric oxide synthase1 (iNOS) 및 cyclooxygenase-2 (COX-2)의 발현 변화를 확인하였다.In order to confirm the anti-inflammatory effect of Example 1 and Comparative Examples 1 to 2, expression of the inflammatory mediators inducible nitric oxide synthase1 (iNOS) and cyclooxygenase-2 (COX-2) by each composition using western blot Changes were confirmed.
구체적으로, raw 264.7 세포를 6 웰 플레이트에 5×105 cells/mL 의 농도로 2700 ㎕씩 분주한 후, 37℃, 5% CO2 incubator에서 24 시간 배양한 뒤 무혈청 배지를 사용하여 18 시간 배양하였다. 각 군을 12.5 ㎍/mL의 농도로 처리한 뒤 2 시간 후에 LPS 1 ㎍/mL를 정상 실험군을 제외한 모든 웰에 처리하였다. Specifically, after dispensing 2700 μl of raw 264.7 cells at a concentration of 5×10 5 cells/mL in a 6-well plate, culturing for 24 hours in a 37°C, 5% CO 2 incubator, and then using a serum-free medium for 18 hours cultured. After each group was treated at a concentration of 12.5 μg/mL, 2 hours later, 1 μg/mL of LPS was applied to all wells except for the normal experimental group.
처리 후 24시간 후 상층액을 제거하고 인산염 완충 식염수(phosphate buffered saline, PBS: Sigma-Aldrich) 로 2회 세척한 뒤 스크레이퍼(scrapper)를 이용하여 세포를 수확하여 세포에 용해 버퍼(lysis buffer, Sigma-Aldrich) 를 웰당 100 ㎕ 씩 첨가한 뒤 1 시간 동안 냉장하여 세포를 용해시킨 후 원심 분리하여 단백질을 수득하였다. 상기 수득한 단백질은 브래드 포드 분석(bradford assay)으로 정량하고 10 ㎕의 단백질을 10% 나트륨도데실설페이트폴리아크릴아미드 겔 전기영동법(sodium dodecyl sulfate polyacrylamide gel electrophoresis)을 이용해 전기영동을 실시한 후 니트로 셀룰로오스 막(nitrocellulose membrane, AmerchamTM, GE Healthcare Life Sciences)에 이동시켰다. After 24 hours of treatment, the supernatant was removed, washed twice with phosphate buffered saline (PBS: Sigma-Aldrich), and then the cells were harvested using a scraper and lysed in lysis buffer (Sigma-Aldrich). -Aldrich) was added at 100 μl per well and refrigerated for 1 hour to lyse the cells, followed by centrifugation to obtain proteins. The obtained protein was quantified by Bradford assay, and 10 μl of the protein was subjected to electrophoresis using 10% sodium dodecyl sulfate polyacrylamide gel electrophoresis, and then the nitrocellulose membrane (nitrocellulose membrane, Amercham TM , GE Healthcare Life Sciences).
다음으로, 상기 단백질을 차단 용액(blocking solution, 5% skim milk in mixture of tris-buffered saline and tween 20 (TBST) buffer] 을 이용하여 24 시간 동안 블락킹(blocking) 한 뒤, 1차 항체(antibody, anti-mouse iNOS, anti-mouse COX2, anti-mouse Actin; Santa Cruz Biotechnology, USA)를 24 시간 반응시킨 후 겨자무과산화수소(horseradish peroxidase: HRP)가 결합된 2차 항체(anti-mouse IgG; Santa Cruz Biotechnology)를 1:1000으로 희석하여 3시간 반응시켰다. Next, after blocking the protein for 24 hours using a blocking solution (5% skim milk in mixture of tris-buffered saline and tween 20 (TBST) buffer), the primary antibody , anti-mouse iNOS, anti-mouse COX2, anti-mouse Actin; Santa Cruz Biotechnology, USA) after reacting for 24 hours, horseradish peroxidase (HRP)-conjugated secondary antibody (anti-mouse IgG; Santa Cruz Biotechnology) was diluted 1:1000 and reacted for 3 hours.
다음으로, TBST로 3회 세척한 뒤 진보된 화학발광법(enhanced chemiluqnsescence: ECL) 키트(AmershamTM)를 사용하여, ECL 기질(AmershamTM)과 반응 후 Chemiluqnsescent Imaging System (LuqnsoGraph; ATTO, Japan)을 이용하여 단백질 발현량을 확인하였다.Next, after washing with TBST three times, using an advanced chemiluqnsescence (ECL) kit (Amersham TM ), after reacting with an ECL substrate (Amersham TM ), a Chemiluqnsescent Imaging System (LuqnsoGraph; ATTO, Japan) The protein expression level was confirmed using .
도 7은 리가다 소나무 껍질 추출물 및 상기 추출물이 봉입된 나노 에토좀의 웨스턴 블랏 결과를 나타낸 도이다.Figure 7 is a diagram showing the Western blot results of Rigada pine bark extract and nano ethosomes encapsulated with the extract.
그 결과, 도 7에서 나타낸 바와 같이 실시예 2를 처리한 경우, 비교군과 비교하여 유의미하게 낮은 염증매개인자 단백질의 발현량을 확인하였다. 이러한 결과는, 일 양상의 조성물이 염증매개인자 단백질의 발현 저해율을 가짐으로써, 항염증 효과가 현저한 것을 의미한다.As a result, as shown in FIG. 7, when Example 2 was treated, it was confirmed that the expression level of the inflammatory mediator protein was significantly lower than that of the control group. These results mean that the anti-inflammatory effect is remarkable because the composition of one aspect has an expression inhibition rate of the inflammatory mediator protein.
실험예 8 : 리기다 소나무 껍질 추출물 봉입 나노 에토좀의 피부 흡수율 실험Experimental Example 8: Skin Absorption Rate Test of Rigida Pine Bark Extract Encapsulated Nano Ethosomes
상기 실시예 1 및 비교예 1의 피부흡수율을 비교하기 위하여, 각 조성물을 피부투과장치(Franz Diffusion Cell)에 고정되어있는 Micropig Franz 세포막에 각각 도포하여 피부 흡수율을 측정하여 누적투과량과 투과속도(Flux, μg/hr*cm2) 및 투과상수(Kp, Permeability coefficient, cm/hr*10-4)를 산출하였다.In order to compare the skin absorption rate of Example 1 and Comparative Example 1, each composition was applied to the Micropig Franz cell membrane fixed to the skin penetration device (Franz Diffusion Cell), and the skin absorption rate was measured to determine the cumulative penetration amount and penetration rate (Flux , μg/hr*cm 2 ) and permeability coefficient (Kp, permeability coefficient, cm/hr*10 -4 ) were calculated.
표 6은 일 양상의 조성물을 처리한 경우, 누적투과량과 투과속도(Flux, μg/hr*cm2) 및 투과상수(Kp, Permeability coefficient, cm/hr*10-4)를 나타낸 것이다.Table 6 shows the cumulative permeation amount and permeability rate (Flux, μg/hr*cm 2 ) and permeability constant (Kp, permeability coefficient, cm/hr*10 -4 ) when the composition of one aspect was treated.
(μg/hr*cm2)Flux
(μg/hr*cm 2 )
(cm/hr*10-4)Kp
(cm/hr*10 -4 )
±286.178148.70
±286.178
±14308.897435.05
±14308.89
±53.51529.00
±53.515
±2675.751450.47
±2675.75
도 8은 프라츠 분산 세포(Franz Diffusion Cell) 실험을 통해 리가다 소나무 껍질 추출물 및 상기 추출물이 봉입된 나노 에토좀의 피부 흡수율을 나타낸 그래프이다.8 is a graph showing the skin absorption rate of Rigada pine bark extract and nano ethosomes encapsulated with the extract through a Franz Diffusion Cell experiment.
그 결과, 도 8에 나타낸 것과 같이, 각 시간대별로 리기다 소나무 껍질 추출물이 봉입된 나노 에토좀이 도포된 실험군에서 높은 흡수율과 빠른 흡수 시간대를 갖는 것을 확인하였다. 특히 최초 도포 후 1시간 경과시에는 비교예 1은 흡수가 되지 않았지만 실시예 1에서는 높은 피부 흡수율을 보였다.As a result, as shown in FIG. 8, it was confirmed that the experimental group coated with nano-ethosomes encapsulated with Rigida pine bark extract for each time period had a high absorption rate and a fast absorption time period. In particular, Comparative Example 1 did not absorb at 1 hour after the first application, but Example 1 showed a high skin absorption rate.
특히, 상기 측정된 누적투과량으로 계산된 투과속도 값에 의하면, 리기다 소나무 껍질 추출물보다 이를 봉입한 나노 에토좀의 투과속도가 약 5배 높은 것으로 확인되며, 투과상수값에 따른 기준도 매우 빠름을 확인하였다. 이러한 결과는, 리기다 소나무 껍질 추출물보다 이를 봉입한 나노 에토좀이 피부 흡수율이 더 높음을 의미한다.In particular, according to the permeation rate value calculated from the measured cumulative permeation amount, it was confirmed that the permeation rate of the nano-ethosome encapsulated therein was about 5 times higher than that of the Rigida pine bark extract, and the standard according to the permeation constant value was also very fast. did These results indicate that the skin absorption rate of the nano-ethosome encapsulated therein is higher than that of the Rigida pine bark extract.
실험예 9 : 리가다 소나무 껍질 추출물 나노 에토좀이 함유된 크림의 피부 가려움증 완화 및 경피수분손실량(TEWL) 개선효과 인체적용시험Experimental Example 9: Skin Itching Relief and Transepidermal Water Loss (TEWL) Improvement Effect of Cream Containing Rigada Pine Bark Extract Nano Ethosome Human Application Test
상기 제조예 1의 크림 화장료 조성물의 피부의 가려움 완화 및 경피 수분 손실량 (TEWL) 개선 효과를 확인하기 위하여 인체 적용 시험을 수행하였다.In order to confirm the effects of the cream cosmetic composition of Preparation Example 1 on relieving skin itchiness and improving transepidermal water loss (TEWL), a human application test was performed.
구체적으로, 만 30 내지 65세 사이의 남녀 피험자 21명을 대상으로 4주간 아침, 저녁 제조예 1을 적당량 덜어 피부 건조와 가려움을 느끼는 부위에 도포한 후 부드럽게 문질러 흡수시킨 후, 실험 진행 후 2주차 및 4주차에 평가하였다. 상기 평가는 Corneometer를 이용히여 피부 건조와 가려움을 느끼는 부위의 피부보습 측정값을 분석하였다. 또한, Tewameter를 이용한 피부 건조와 가려움을 느끼는 부위의 경피수분손실량(TEWL)측정값을 분석하였다.Specifically, for 21 male and female subjects between the ages of 30 and 65, take an appropriate amount of Preparation Example 1 in the morning and evening for 4 weeks, apply it to the area where the skin feels dry and itchy, rub it gently, and absorb it. and 4 weeks. The evaluation was performed using a corneometer to analyze skin moisture measurement values in areas where skin dryness and itchiness were felt. In addition, the measured values of transepidermal water loss (TEWL) in areas where dryness and itchiness were felt using a Tewameter were analyzed.
표 7은 일 양상의 조성물을 도포한 경우의 피부보습 증가율 및 경피수분손실량 감소율을 나타낸 것이다.Table 7 shows the rate of increase in skin moisturization and decrease in transepidermal water loss when the composition of one aspect was applied.
그 결과, 표 7에 나타낸 것과 같이, 피부보습 측정값은 제조예 1 적용 전에 비하여 적용 2주 후 및 적용 4주 후 통계적으로 유의한 수준(p<0.001)으로 증가하였으며, 각각 4.680%, 7.051%의 증가율을 나타냈다. 또한, 경피수분손실률은 제조예 1 적용 전에 비하여 적용 2주 후 및 적용 4주 후 통계적으로 유의한 수준(p<0.01)으로 감소하였으며, 각각 5.400%, 7.480%의 감소율을 나타내었다. 이러한 결과는 일 양상의 조성물이 경피수분손실을 감소시키고 및 피부보습효과가 높아 건조한 피부의 가려움 완화에 도움을 주는 것을 의미한다.As a result, as shown in Table 7, the measured skin moisturizing value increased at a statistically significant level (p<0.001) after 2 weeks and 4 weeks after application compared to before application of Preparation Example 1, respectively, 4.680% and 7.051% showed an increase in In addition, the transepidermal water loss rate decreased at a statistically significant level (p<0.01) after 2 weeks and 4 weeks after application compared to before application of Preparation Example 1, respectively, showing a reduction rate of 5.400% and 7.480%. These results mean that the composition of one aspect reduces transepidermal water loss and has a high skin moisturizing effect, helping to relieve itchiness of dry skin.
실험예 10 : 리가다 소나무 껍질 추출물 나노 에토좀이 함유된 크림의 피부 진정 인체적용시험Experimental Example 10: Skin soothing human body application test of cream containing rigada pine bark extract nano ethosomes
상기 제조예 1의 크림 화장료 조성물의 피부 진정 효과를 확인하기 위하여 인체 적용 시험을 수행하였다.In order to confirm the skin soothing effect of the cream cosmetic composition of Preparation Example 1, a human application test was performed.
구체적으로, 만 20 내지 60세의 여성 피험자 22명을 대상으로 시험부위에 1% SLS(Sodium Lauryl Sulfate) 용액을 IQ chamber와 filter paper를 이용하여 24시간 동안 폐쇄첩포하여 장벽 손상을 유도하였으며, 시험부위(제품 적용 부위 및 미적용 부위)는 연구 대상자의 손목으로부터 10 ~ 15cm 거리의 전완부 부위로 하고 크기는 1cm2으로 하였다. 다음으로, 14일간 1일 아침, 저녁으로 제조예 1을 제품 적용 부위에 적당량 도포하여 흡수시켰다. 이후 1일, 2일, 6일 및 14일 간 제품 적용 전, 적용 후 모두 측정을 진행하였다.Specifically, 22 female subjects between the ages of 20 and 60 were closed for 24 hours with 1% SLS (Sodium Lauryl Sulfate) solution on the test site using an IQ chamber and filter paper to induce barrier damage. The area (product applied area and non-applied area) was the forearm area at a distance of 10 to 15 cm from the subject's wrist and the size was 1 cm 2 . Next, Preparation Example 1 was applied in an appropriate amount to the product application site in the morning and evening for 14 days and absorbed. After that, measurements were carried out before and after application of the product for 1 day, 2 days, 6 days, and 14 days.
도 9는 리기다 소나무 껍질 추출물을 봉입한 나노 에토좀이 포함된 크림의 피부 진정 효과를 나타낸 이미지이다.9 is an image showing the skin soothing effect of a cream containing nano-ethosomes encapsulated with Rigida pine bark extract.
표 8은 일 양상의 조성물을 도포한 경우의 피부 홍반도 감소율 및 경피수분손실량 감소율을 나타낸 것이다.Table 8 shows the reduction rate of skin erythema and the reduction rate of transepidermal water loss when the composition of one aspect was applied.
(%)Decreased rate of transepidermal water loss
(%)
(%)Skin erythema reduction rate
(%)
그 결과, 도 9 및 표 8에 나타낸 것과 같이, 제조예 1을 도포한 부위는 미적용 부위에 비해 통계적으로 유의한 수준(p<0.01)으로 높은 경피수분손실량 개선율을 보였으며, 14일차에서 92.596%의 개선율을 나타냈다. 또한, M제조예 1을 도포한 부위는 미적용 부위에 비해 통계적으로 유의한 수준 (p<0.05)으로 낮은 피부 홍반도를 보였으며, 14일차에서 19.537%의 감소율을 나타내었음을 확인하였다. 이러한 결과는, 일 양상의 조성물이 피부 진정 효과가 뛰어난 것을 의미하는 것이다.As a result, as shown in FIG. 9 and Table 8, the area to which Preparation Example 1 was applied showed a statistically significant level (p<0.01) of high improvement in transepidermal water loss compared to the unapplied area, and 92.596% on the 14th day. showed an improvement rate of In addition, it was confirmed that the area where M Preparation Example 1 was applied showed a statistically significant level (p <0.05) of low skin erythema compared to the unapplied area, and showed a reduction rate of 19.537% on the 14th day. These results mean that the composition of one aspect has an excellent skin soothing effect.
Claims (11)
상기 혼합액을 정제수와 교반하여 에토좀(Ethosome)을 제조하는 단계를 포함하는, 피부 진정, 피부 장벽 재생, 피부 간지러움, 피부 홍반 개선 또는 예방용 화장료 제조 방법.Rigida pine bark extract (Pinus rigida Mill. bark extract) or a fraction thereof (Fraction) and phospholipids to prepare a mixed solution; and
A cosmetic preparation method for soothing skin, regenerating skin barrier, improving or preventing skin itching, skin erythema, comprising the step of preparing ethosome by stirring the mixed solution with purified water.
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| KR20150078948A (en) * | 2013-12-31 | 2015-07-08 | 주식회사 삼양제넥스 | Cosmetic composition for improving skin inflammation or reducing skin irritation |
| KR20190137327A (en) * | 2018-06-01 | 2019-12-11 | 애경산업(주) | Skin external application composition for anti-aging |
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| KR20150078948A (en) * | 2013-12-31 | 2015-07-08 | 주식회사 삼양제넥스 | Cosmetic composition for improving skin inflammation or reducing skin irritation |
| KR20190137327A (en) * | 2018-06-01 | 2019-12-11 | 애경산업(주) | Skin external application composition for anti-aging |
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