KR102059392B1 - Antimicrobial and Antifungal Composition Containing Clove, Hibiscus, and Coconut Extract Complex as Active Ingredient - Google Patents
Antimicrobial and Antifungal Composition Containing Clove, Hibiscus, and Coconut Extract Complex as Active Ingredient Download PDFInfo
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- KR102059392B1 KR102059392B1 KR1020180058651A KR20180058651A KR102059392B1 KR 102059392 B1 KR102059392 B1 KR 102059392B1 KR 1020180058651 A KR1020180058651 A KR 1020180058651A KR 20180058651 A KR20180058651 A KR 20180058651A KR 102059392 B1 KR102059392 B1 KR 102059392B1
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- hibiscus
- composition
- clove
- extract
- antimicrobial
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Abstract
본 발명은 정향, 히비스커스, 및 코코넛 추출물을 유효성분으로 포함하는 항균 및 항진균 조성물에 관한 것이다.
본 발명에 따른 정향, 히비스커스, 및 코코넛의 복합 추출물은 유해 미생물에 직접 영향을 미치는 것을 통해 탁월한 항균 및 항진균 활성을 나타내는 동시에 피부 주름개선, 피부 염증 개선, 항산화 등의 효과가 있으므로 약학적 조성물, 화장료 조성물, 식품 또는 사료 보존용 방부제 등의 용도로 유용하게 사용될 수 있다.The present invention relates to an antibacterial and antifungal composition comprising clove, hibiscus, and coconut extract as an active ingredient.
The complex extract of clove, hibiscus, and coconut according to the present invention exhibits excellent antibacterial and antifungal activity through direct effects on harmful microorganisms, and at the same time has an effect of improving skin wrinkles, improving skin inflammation, and antioxidants. It can be usefully used for the purpose of the composition, food or feed preservatives.
Description
본 발명은 항균 및 항진균 활성을 갖는 조성물에 관한 것으로, 더욱 구체적으로 정향, 히비스커스, 및 코코넛 추출물을 유효성분으로 포함하는 항균 및 항진균 조성물에 관한 것이다.The present invention relates to a composition having antibacterial and antifungal activity, and more particularly to an antibacterial and antifungal composition comprising clove, hibiscus, and coconut extract as an active ingredient.
생물체의 항상성(homeostasis)을 유지하기 위한 과정에서 중요한 역할을 담당하고 있는 물질들 중 일부가 각종 생물체 유래의 생리 활성 물질이다. 지금까지 수많은 생리 활성 물질에 대해 많은 연구가 진행되고 있으며, 그 중, 미생물, 식물, 또는 동물체 등에서 분리된 항균물질에 관한 연구는 생명과학 및 의학 분야에서 매우 중요한 영역이다.Some of the substances that play an important role in the process of maintaining homeostasis are biologically active substances from various organisms. Many studies have been conducted on a number of bioactive substances, and among them, research on antimicrobial substances isolated from microorganisms, plants, or animals is a very important area in the life sciences and medicine fields.
모든 생명체는 생존을 위하여 항균 및 항진균 물질을 생산하는 것으로 알려져 있는데, 항균 및 항진균 물질을 생산하는 미생물에 대해서 많은 연구가 이루어지고 있으며, 항균 및 항진균 물질을 생산하는 미생물 또는 이들에게서 분리된 물질을 이용해 항균제 또는 항진균제를 개발하려는 연구들이 오래전부터 시도되고 있다. All living organisms are known to produce antimicrobial and antifungal substances for survival. Much research has been conducted on microorganisms that produce antimicrobial and antifungal substances. Research into the development of antibacterial or antifungal agents has long been attempted.
항균물질은 미생물을 죽이면서 인체 또는 동물에게는 독성이 낮고 체내의 효소 등에 의해 비활성화되지 않는 선택적 독성작용(selective toxicity)을 갖는 물질로, 이는 주로 DNA의 복제, 유전정보의 전사 및 해독, 전자에너지의 수송, 세포벽의 생합성 등을 저해함으로써 미생물의 증식을 억제하는 기전을 통해 효과를 나타낸다.Antimicrobial substances have a selective toxicity that kills microorganisms and are not toxic to humans or animals and are not inactivated by enzymes in the body. This is mainly due to DNA replication, transcription and translation of genetic information, and It inhibits transport, biosynthesis of cell walls, and the like, through the mechanism of inhibiting the growth of microorganisms.
현재까지 개발된 주된 항균물질들은 미생물, 식물 등 천연에서 유래한 것과 화학적으로 합성된 것으로 나뉠 수 있는데, 천연에서 분리된 최초의 항균물질은 1928년 알렉산더 플레밍에 의해 발견된 페니실린이며, 이후 천연 유래 또는 화학적으로 합성된 많은 항균물질이 유해균에 의해 유발된 질병의 치료에 사용되고 있으나, 최근에는 화학적으로 합성된 항균물질에 대한 내성 증가로 인해 천연에서 유래한 항균물질을 이용한 항균제 개발에 관심이 증가하는 추세이다. The main antimicrobial agents developed to date can be divided into microorganisms, plants, etc., derived from nature and chemically synthesized.The first antimicrobial substance isolated from nature was penicillin, discovered by Alexander Fleming in 1928, and then Many chemically synthesized antimicrobials are used for the treatment of diseases caused by harmful bacteria, but recently, due to increased resistance to chemically synthesized antimicrobial agents, interest in developing antimicrobials using natural antimicrobial agents is increasing. to be.
천연 유래 항균 또는 항진균제와 관련된 기술로, 고삼 추출물(특허출원 제10-1993-0006319호), 자몽 추출물(특허출원 제10-1993-0006320호), 회향, 대회향, 세신, 녹나무속 식물 및 정향의 혼합 추출물(특허출원 제10-2002-0054685호), 녹차 폴리페놀 및 티트리 오일(특허출원 제10-2002-0028517호), 세스바니아 그랜디프로라 추출물(특허출원 제10-2011-0015252호) 등 다양한 식물체의 추출물 또는 그 분획물이 항균 활성을 갖는 것으로 지금까지 보고되어 있다. As a technology related to antibacterial or antifungal agents derived from nature, Ginseng extract (Patent Application No. 10-1993-0006319), Grapefruit Extract (Patent Application No. 10-1993-0006320), Fennel, Conventional Fragrance, Sessin, Camphor and Clove Mixed Extract (Patent Application No. 10-2002-0054685), Green Tea Polyphenol and Tea Tree Oil (Patent Application No. 10-2002-0028517), Sesbania Grandiprora Extract (Patent Application No. 10-2011-0015252) It has been reported so far that extracts or fractions of these various plants have antimicrobial activity.
이러한 연구결과에도 불구하고 단일 식물추출물 또는 복합 식물추출물을 항균 및 항진균 용도로 사용하고자 하는 시도가 있었으나, 아직 산업 현장에서 활용되기에 충분할 만큼의 항균 또는 항진균 효과를 나타내는 식물추출물은 거의 없으며, 항균 및 항진균제와 관련된 연구 성과는 미미한 상태이다. 또한, 항균제로서의 고유 기능 이외에 피부주름 개선, 항염 활성 등 항균제 자체가 보유한 기능성에 이외의 다양한 부가 효능에 관한 연구는 이루어진 바 없다. Despite these findings, attempts have been made to use single or multiple plant extracts for antibacterial and antifungal applications, but few plant extracts exhibit sufficient antimicrobial or antifungal effects to be used in industrial sites. Research results related to antifungal drugs are minimal. In addition, in addition to the intrinsic function as an antimicrobial agent, research on various additional effects other than the functionality possessed by the antimicrobial agent itself such as skin wrinkle improvement and anti-inflammatory activity has not been made.
이러한 배경 아래에서, 본 발명자들은 우수한 항균 및 항진균 활성을 갖는 동시에 자체적으로 피부주름 개선, 항염 활성 등의 다양한 기능성을 보유한 식물추출물들의 조합을 찾기 위해 예의 노력한 결과, 정향, 히비스커스, 및 코코넛 추출물의 복합 추출물이 다양한 유해 미생물들을 효과적으로 억제하여 항균제 및 항진균제로 유용하게 활용될 수 있음을 확인하고, 본 발명을 완성하게 되었다.Under this background, the present inventors have diligently sought to find a combination of plant extracts having excellent antibacterial and antifungal activity and at the same time having various functionalities such as skin wrinkle improvement and anti-inflammatory activity, resulting in a combination of clove, hibiscus, and coconut extract. It was confirmed that the extract can be effectively used as an antibacterial and antifungal agent by effectively inhibiting various harmful microorganisms, and completed the present invention.
따라서, 본 발명의 주된 목적은 다양한 유해 미생물들을 효과적으로 억제하여 항균제 및 항진균제로 유용하게 활용될 수 있는 정향, 히비스커스, 및 코코넛의 복합 추출물을 유효성분으로 포함하는 조성물을 제공하는 데 있다.Therefore, the main object of the present invention is to provide a composition comprising a complex extract of cloves, hibiscus, and coconut as an active ingredient that can effectively inhibit various harmful microorganisms, which can be usefully used as an antibacterial and antifungal agent.
본 발명의 다른 목적 및 이점은 하기의 발명의 상세한 설명, 청구범위 및 도면에 의해 더욱 명확하게 된다.Other objects and advantages of the present invention will become apparent from the following detailed description, claims and drawings.
본 발명의 한 양태에 따르면, 본 발명은 정향, 히비스커스, 및 코코넛 추출물을 유효성분으로 포함하는 항균 및 항진균 조성물을 제공한다.According to one aspect of the invention, the present invention provides an antibacterial and antifungal composition comprising clove, hibiscus, and coconut extract as an active ingredient.
본 발명자들은 수십 종의 식물추출물들의 조합을 검색하여 우수한 항균 및 항진균 효능이 갖는 물질을 탐색한 결과, 정향, 히비스커스, 및 코코넛 추출물의 조합이 항균 및 항진균 효과가 획기적으로 증가함을 발견하였고, 또한 피부에 적용시에도 안전성에도 문제가 없음을 확인하였다.The present inventors searched for a combination of dozens of plant extracts and searched for substances having excellent antibacterial and antifungal efficacy, and found that the combination of cloves, hibiscus, and coconut extract significantly increased the antibacterial and antifungal effect. It was confirmed that there is no problem in safety even when applied to the skin.
본 발명의 "정향"은 학명 유제니아 카리오필라타(Eugenia caryophyllatum)로 분류된 식물을 의미하고, 용어 "히비스커스(Hibiscus syriacus)"는 한반도 전역에서 자생하는 쌍떡잎식물 아욱목 아욱과의 낙엽관목을 의미하며, 용어 "코코넛(Cocos nucifera L.)"은 남미 원산의 야자과 식물인 야자의 과실을 의미한다. The term "clove" of the present invention means a plant classified under the scientific name Eugenia caryophyllatum, and the term "Hibiscus syriacus" means a deciduous shrub of the dicotyledonous mallow plant native to the whole Korean peninsula. The term “cocos (Cocos nucifera L.)” refers to the fruit of a palm, a palm plant native to South America.
본 발명에서 사용되는 용어 "추출물"은 식물의 추출 처리에 따라 얻어지는 추출액, 상기 추출액의 희석액이나 농축액, 상기 추출액을 건조하여 얻어지는 건조물, 상기 추출액의 조정제물이나 정제물, 또는 이들의 혼합물 등, 추출액 자체 및 추출액을 이용하여 형성 가능한 모든 제형의 추출물을 포함한다. 본 발명의 상기 추출물은 바람직하게 추출 후 건조 분말 형태로 제조되어 사용될 수 있다. The term "extract" as used in the present invention is an extract, such as an extract obtained by the extraction process of a plant, a diluent or concentrate of the extract, a dried product obtained by drying the extract, a crude or purified product of the extract, or a mixture thereof. Extracts of all formulations that can be formed using itself and extracts. The extract of the present invention may be preferably prepared and used in the form of a dry powder after extraction.
본 발명의 상기 정향, 히비스커스, 및 코코넛 추출물은, 천연, 잡종 또는 변종 식물로부터 추출될 수 있고, 식물 조직 배양물에서도 추출할 수 있으며, 잎, 줄기, 뿌리, 꽃, 열매 등의 부위에서 추출한 추출물을 포함한다. The clove, hibiscus, and coconut extract of the present invention can be extracted from natural, hybrid or variety plants, can also be extracted from plant tissue culture, extracts extracted from the leaves, stems, roots, flowers, fruit, etc. It includes.
본 발명의 항균 및 항진균 조성물에서, 상기 정향, 히비스커스, 및 코코넛은 1:8:9의 중량비로 조합되는 것이 가장 바람직하다.In the antimicrobial and antifungal compositions of the invention, the clove, hibiscus, and coconut are most preferably combined in a weight ratio of 1: 8: 9.
본 발명의 실시예에서 상기 정향, 히비스커스, 및 코코넛을 다양한 비율로 혼합하여 항균 및 항진균 조성물을 제조하였으며, 제조된 조성물의 항균력을 확인하기 위해 한천확산법(disc diffusion test) 및 미생물 생장 최소억제 농도 평가를 수행하였다. 그 결과, 정향, 히비스커스, 및 코코넛의 조합이 전반적으로 우수한 항균 활성을 나타내는 것을 확인하였으며, 특히 대조군으로 사용한 메틸파라벤과 자몽종자 추출물와 비교하였을 때 동등 이상의 항균 효과를 나타내어, 상기 정향, 히비스커스, 및 코코넛 추출물이 1:8:9의 비율로 혼합된 조성물이 가장 우수한 시너지 효과가 있는 것을 확인할 수 있었다.In the embodiment of the present invention, the clove, hibiscus, and coconut were mixed in various ratios to prepare an antimicrobial and antifungal composition, and to confirm the antimicrobial activity of the prepared composition, agar diffusion (disc diffusion test) and microbial growth inhibition concentration evaluation Was performed. As a result, it was confirmed that the combination of clove, hibiscus, and coconut showed an excellent antimicrobial activity as a whole, especially when compared to the methylparaben and grapefruit seed extract used as a control group, showed the same or more antibacterial effect, the clove, hibiscus, and coconut It was confirmed that the composition mixed with the extract in a ratio of 1: 8: 9 has the best synergistic effect.
본 발명의 항균 및 항진균 조성물은 넓은 항균 스펙트럼을 갖는데, 구체적으로 그람양성균, 그람음성균, 효모, 및 진균에 항균 활성을 갖는다.The antimicrobial and antifungal compositions of the present invention have a broad antimicrobial spectrum, specifically, antimicrobial activity against Gram-positive bacteria, Gram-negative bacteria, yeast, and fungi.
본 발명의 조성물은 구체적으로는 칸디다 속 진균과 아스퍼질러스 속 진균에 대해 항진균 활성을 갖고, 보다 구체적으로는 칸디다 알비칸스(Candida albicans), 및 아스퍼질러스 니거(Aspergillus niger)에 대해 항진균 활성이 있으나, 이에 한정되는 것은 아니다. Specifically, the composition of the present invention has antifungal activity against Candida fungi and Aspergillus fungi, and more specifically, antifungal activity against Candida albicans and Aspergillus niger. However, the present invention is not limited thereto.
또한, 본 발명의 조성물은 구체적으로 에스체리치아(Escherichia) 속 균주, 및 슈도모나스(Pseudomonas) 속의 그람음성 균주에 대해 항균 활성을 갖고, 보다 구체적으로 에스체리치아 콜라이(Escherichia coli), 및 슈도모나스 에루지노사(Pseudomonas aeruginosa) 균주에 대해 항균 활성이 있으나, 이에 한정되는 것은 아니다.In addition, the composition of the present invention has antimicrobial activity specifically against Escherichia sp. Strains, and Gram-negative strains of Pseudomonas sp., More specifically Escherichia coli, and Pseudomonas aeruginosa. There is antibacterial activity against Pseudomonas aeruginosa strains, but is not limited thereto.
또한, 본 발명의 조성물은 구체적으로 바실러스(Bacillus) 속 균주, 및 스타필로코커스(Staphylococcus) 속의 그람양성 균주에 대해 항균 활성을 갖고, 보다 구체적으로 스타필로코커스 아우레우스(Staphylococcus aureus), 바실러스 서브틸리스(Bacillus subtilis) 등의 세균에 대하여 항균 활성이 있으나, 이에 한정되는 것은 아니다.In addition, the composition of the present invention has an antimicrobial activity against Bacillus sp. Strain, and Gram-positive strains of the Staphylococcus spp., More specifically Staphylococcus aureus, Bacillus sub Antibacterial activity against bacteria such as Bacillus subtilis, but is not limited thereto.
본 발명의 실시예에서는 정향, 히비스커스, 및 코코넛 복합 추출물이 갖는 항균 효과를 검정한 결과, 이들의 조합이 각각의 단일 화합물과 비교해 매우 우수한 항균 활성을 갖는 것을 확인하였다.In the examples of the present invention, the antibacterial effect of the clove, hibiscus, and coconut complex extract was assayed. As a result, it was confirmed that the combination thereof had a very good antimicrobial activity compared to each single compound.
한편, 피부섬유아세포의 콜라겐은 피부 재생, 피부 탄력, 피부 주름 형성 및 피부 손상 시 조직의 수복 또는 재생과 직접적인 관련이 있다고 볼 수 있다.On the other hand, collagen of dermal fibroblasts can be said to be directly related to the repair or regeneration of tissue during skin regeneration, skin elasticity, skin wrinkle formation and skin damage.
즉, 피부 섬유아세포의 콜라겐 또는 프로콜라겐의 합성이 촉진되거나, 콜라겐을 분해하는 기질 단백질 분해효소(matrix metallo-proteinase, 이하 'MMP'라 함)의 합성이 저해되면, 피부 탄력 개선, 피부 재생, 피부 주름 개선, 상처 치유, 손상된 피부 조직의 수복 및 재생; 및 피부 노화 방지 등의 효과를 얻을 수 있다.That is, when the synthesis of collagen or procollagen of skin fibroblasts or the synthesis of matrix metallo-proteinase (hereinafter referred to as 'MMP') that breaks down collagen is inhibited, skin elasticity improvement, skin regeneration, Skin wrinkle improvement, wound healing, repair and regeneration of damaged skin tissue; And skin aging prevention.
본 발명의 정향, 히비스커스, 및 코코넛 복합 추출물은 MMP-1 단백질의 발현을 억제하는 것을 특징으로 한다.Clove, hibiscus, and coconut complex extract of the present invention is characterized by inhibiting the expression of MMP-1 protein.
구체적으로, 본 발명의 정향, 히비스커스, 및 코코넛 복합 추출물을 유효성분으로 하는 조성물은 자외선 조사된 정상 인간 진피 섬유아세포(NHDF)에서 세포 독성을 저감시키고, 콜라겐 분해를 촉진시키는 기질 단백질 분해효소-1(matrix metallo-proteinase)의 발현을 억제시키는바, 피부 탄력 증가, 피부 재생, 피부 주름 개선, 상처 치유, 손상된 피부 조직의 수복 및 재생; 및 피부 노화 방지 등의 효과 등을 가질 수 있다. 구체적으로는 피부주름의 예방 또는 개선의 효과를 가질 수 있다(도 1 참조).Specifically, the composition comprising the extract of the clove, hibiscus, and coconut complex of the present invention as an active ingredient is a substrate protease-1 which reduces cytotoxicity and promotes collagen degradation in normal human dermal fibroblasts (NHDF) irradiated with ultraviolet rays. inhibits the expression of matrix metallo-proteinases such as increased skin elasticity, skin regeneration, skin wrinkle improvement, wound healing, repair and regeneration of damaged skin tissue; And it can have effects, such as anti-aging of skin. Specifically, it may have an effect of preventing or improving skin wrinkles (see FIG. 1).
또한, 본 발명의 정향, 히비스커스, 및 코코넛 복합 추출물을 유효성분으로 하는 조성물은 IL-6 단백질의 발현을 억제하는 것을 특징으로 한다(도 2 참조). 손상된 피부는 염증반응을 통해 더욱 확대되며, 피부 발적, 피부 과민반응, 또는 광과민 반응 등을 통해 임상적으로 피부가 더욱 악화되는 경향을 보인다. 따라서, 손상된 피부 상태의 효과적인 개선을 위해서 염증반응의 조기 차단은 매우 중요하다.In addition, the composition of the clove, hibiscus, and coconut complex extract of the present invention as an active ingredient is characterized by inhibiting the expression of IL-6 protein (see Fig. 2). Damaged skin is further enlarged through inflammatory reactions, and skin tends to be worsened clinically through skin redness, skin hypersensitivity, or photosensitization. Therefore, early blocking of inflammatory response is very important for effective improvement of damaged skin condition.
본 발명의 바람직한 실시예에서는, 상기 정향, 히비스커스, 및 코코넛 복합 추출물을 처리시 염증 매개 물질인 IL-6 단백질의 생성이 억제되는 것으로 나타났으며, 이러한 IL-6 단백질 발현에 대한 억제능은 농도 의존적인 것으로 나타났다. 따라서, 본 발명의 조성물은 염증매개 물질인 IL-6 단백질의 생성을 효과적으로 억제하고, 피부 주변의 염증반응을 효과적으로 차단하여 손상된 피부의 상태개선에 효과적으로 작용한다.In a preferred embodiment of the present invention, the treatment of the clove, hibiscus, and coconut complex extract has been shown to inhibit the production of IL-6 protein, an inflammation mediator, and the inhibitory ability of the IL-6 protein expression is concentration dependent. Appeared to be. Therefore, the composition of the present invention effectively inhibits the production of the IL-6 protein, an inflammation mediator, and effectively blocks the inflammatory response around the skin, thereby effectively acting to improve the condition of the damaged skin.
또한, 본 발명의 정향, 히비스커스, 및 코코넛의 복합추출물은 우수한 항산화 효과를 나타내며, 이러한 항산화 효과는 양성대조군인 알부틴과 비교할 때, 더 우수한 것으로 나타났다(도 3 참조).In addition, the complex extract of the clove, hibiscus, and coconut of the present invention shows an excellent antioxidant effect, this antioxidant effect was found to be superior when compared to the arbutin positive control group (see Figure 3).
본 발명의 다른 양태에 따르면, 본 발명은 상기 정향, 히비스커스, 및 코코넛 추출물을 유효성분으로 포함하는 항균 및 항진균용 약학적 조성물을 제공한다.According to another aspect of the present invention, the present invention provides a pharmaceutical composition for antibacterial and antifungal comprising the clove, hibiscus, and coconut extract as an active ingredient.
본 발명의 정향, 히비스커스, 및 코코넛의 복합 식물추출물은 유해 미생물에 직접 영향을 미치는 것을 통해 탁월한 항균 및 항진균 활성을 나타내므로 항균 및 항진균용 약학적 조성물로 유용하게 사용될 수 있다.The complex plant extract of clove, hibiscus, and coconut of the present invention exhibits excellent antibacterial and antifungal activity through direct effects on harmful microorganisms, and thus can be usefully used as a pharmaceutical composition for antibacterial and antifungal diseases.
한편, 본 발명의 정향, 히비스커스, 및 코코넛 추출물을 유효성분으로 포함하는 항균 및 항진균용 조성물은 약학적으로 허용 가능한 염의 형태로 제조될 수 있다. 구체적으로 산을 첨가함으로써 염을 형성할 수 있고, 예를 들어 무기산(예: 염산, 히드로브롬산, 인산, 질산, 황산 등), 유기 카르복실산(예: 아세트산, 트리플루오로아세트산과 같은 할로 아세트산, 프로피온산, 말레산, 숙신산, 말산, 시트르산, 타르타르산, 살리실산), 및 산성 당(글루쿠론산, 갈락투론산, 글루콘산, 아스코르브산), 산성 폴리사카리드(예: 히알우론산, 콘드로이틴 술페이트, 아르기닌산), 콘드로이틴 술페이트와 같은 술폰산 당 에스테르를 포함하는 유기 술폰산(예: 메탄, 술폰산, p-톨루엔 술폰산) 등을 첨가하여 염을 형성할 수 있다.On the other hand, antimicrobial and antifungal compositions comprising clove, hibiscus, and coconut extract of the present invention as an active ingredient may be prepared in the form of a pharmaceutically acceptable salt. Specifically, salts can be formed by the addition of acids, for example inorganic acids (e.g. hydrochloric acid, hydrobromic acid, phosphoric acid, nitric acid, sulfuric acid, etc.), organic carboxylic acids (e.g. acetic acid, trifluoroacetic acid) Acetic acid, propionic acid, maleic acid, succinic acid, malic acid, citric acid, tartaric acid, salicylic acid), and acidic sugars (glucuronic acid, galacturonic acid, gluconic acid, ascorbic acid), acidic polysaccharides (e.g. hyaluronic acid, chondroitin sulfate, Salts can be formed by addition of organic sulfonic acids (eg, methane, sulfonic acid, p-toluene sulfonic acid), including sulfonic acid sugar esters such as arginine acid), chondroitin sulfate.
본 발명의 정향, 히비스커스, 및 코코넛 추출물을 유효성분으로 포함하는 항균 및 항진균용 조성물은 천연 식물추출물로 임상투여시 경구 또는 비경구로 투여가 가능하며 일반적인 의약품 제제의 형태로 사용될 수 있다.Antimicrobial and antifungal compositions comprising clove, hibiscus, and coconut extract of the present invention as active ingredients can be administered orally or parenterally during clinical administration with natural plant extracts and can be used in the form of general pharmaceutical formulations.
즉, 본 발명의 정향, 히비스커스, 및 코코넛 추출물을 유효성분으로 포함하는 항균 및 항진균용 조성물은 실제로 경구 또는 비경구의 여러 가지 제형으로 투여될 수 있는데, 제제화할 경우에는 보통 사용하는 충진제, 증량제, 결합제, 습윤제, 붕해제, 계면활성제 등의 희석제 또는 부형제를 사용하여 조제된다. 비경구투여를 위한 제제에는 멸균된 수용액, 비수성용제, 현탁제, 유제, 동결건조제제, 좌제가 포함된다. 비수성용제, 현탁용제로는 프로필렌글리콜(Propylene glycol), 폴리에틸렌 글리콜, 올리브 오일과 같은 식물성 기름, 에틸올레이트와 같은 주사 가능한 에스테르 등이 사용될 수 있다. 좌제의 기제로는 위텝솔(witepsol), 마크로골, 트윈(tween) 61, 카카오지, 리우린지, 글리세로제라틴 등이 사용될 수 있다.That is, the antimicrobial and antifungal composition comprising the clove, hibiscus, and coconut extract of the present invention as an active ingredient may be administered in various oral or parenteral formulations, and when formulated, commonly used fillers, extenders, and binders. And diluents or excipients such as wetting agents, disintegrating agents and surfactants. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, suppositories. As the non-aqueous solvent and the suspension solvent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like can be used. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, uririnji, glycerogelatin and the like can be used.
또한, 본 발명의 정향, 히비스커스, 및 코코넛 추출물을 유효성분으로 포함하는 항균 및 항진균용 조성물은 생리식염수 또는 유기용매와 같이 약제로 허용된 여러 전달체(carrier)와 혼합하여 사용될 수 있고, 안정성이나 흡수성을 증가시키기 위하여 글루코스, 수크로스 또는 덱스트란과 같은 카보하이드레이트, 아스코르브 산(ascorbic acid) 또는 글루타치온과 같은 항산화제(antioxidants), 킬레이트화제(chelating agents), 저분자 단백질 또는 다른 안정화제(stabilizers)들이 약제로 사용될 수 있다.In addition, the antimicrobial and antifungal composition comprising the clove, hibiscus, and coconut extract of the present invention as an active ingredient can be used in combination with various carriers (pharmaceutically acceptable carrier) such as physiological saline or organic solvent, stability or absorbency Carbohydrates such as glucose, sucrose or dextran, antioxidants such as ascorbic acid or glutathione, chelating agents, small molecule proteins or other stabilizers are used to increase Can be used as
본 발명의 정향, 히비스커스, 및 코코넛 추출물을 유효성분으로 포함하는 항균 및 항진균용 조성물의 유효용량은 0.01 내지 10 ㎎/㎏이고, 바람직하게는 0.1 내지 1 ㎎/㎏ 이며, 하루 1회 내지 3회 투여될 수 있다.The effective dose of the antimicrobial and antifungal composition comprising clove, hibiscus, and coconut extract of the present invention as an active ingredient is 0.01 to 10 mg / kg, preferably 0.1 to 1 mg / kg, once to three times a day. May be administered.
본 발명의 약학적 조성물에서 또는 그 배양액은 총 유효량은 볼루스(bolus) 형태 혹은 상대적으로 짧은 기간 동안 주입(infusion) 등에 의해 단일 투여량(single does)으로 환자에게 투여될 수 있으며, 다중 투여량(multiple does)이 장기간 투여되는 분할 치료 방법(fractionated treatment protocol)에 의해 투여될 수 있다. 상기 농도는 약의 투여 경로 및 치료 횟수뿐만 아니라 환자의 나이 및 건강상태 등 다양한 요인들을 고려하여 환자의 유효 투여량이 결정되는 것이므로 이러한 점을 고려할 때, 이 분야의 통상적인 지식을 가진 자라면 본 발명의 정향, 히비스커스, 및 코코넛 복합 식물추출물의 약학적 조성물로서의 특정한 용도에 따른 적절한 유효 투여량을 결정할 수 있을 것이다.In the pharmaceutical composition of the present invention or the culture medium thereof, the total effective amount may be administered to the patient in a single dose by bolus form or by infusion for a relatively short period of time, and the like. Multiple does may be administered by a fractionated treatment protocol with long term administration. Since the concentration is determined by taking into consideration various factors such as the age and health condition of the patient as well as the route of administration and the number of treatments of the drug, in view of the above, the present invention is a person having ordinary skill in the art. Appropriate effective dosages for the specific use of the clove, hibiscus, and coconut complex plant extract as pharmaceutical compositions may be determined.
본 발명의 또 다른 양태에 따르면, 본 발명은 정향, 히비스커스, 및 코코넛 추출물을 유효성분으로 포함하는 항균 및 항진균용 화장료 조성물을 제공한다.According to another aspect of the present invention, the present invention provides a cosmetic composition for antibacterial and antifungal comprising clove, hibiscus, and coconut extract as an active ingredient.
본 발명의 정향, 히비스커스, 및 코코넛 추출물을 유효성분으로 포함하는 항균 및 항진균용 화장료 조성물은 천연유래 식물 소재에서 추출되어 인체에 해가 없고, 화장품이 유해 미생물에 의해 부패하는 것을 효과적으로 막아주며, 그 첨가로 인해 화장품의 물질의 품질을 손상시키지 않으므로 화장료 조성물에 포함되어 사용되는데 적합하고, 특히 피부주름개선, 피부염증 완화, 및 항산화 효과가 있어 화장료 조성물에 포함되어 사용되는 것이 바람직하다.The antimicrobial and antifungal cosmetic composition comprising the clove, hibiscus, and coconut extract of the present invention as an active ingredient is extracted from natural plant material, which is harmless to the human body, and effectively prevents the cosmetic from being corrupted by harmful microorganisms, Since the addition does not impair the quality of the material of the cosmetic, it is suitable to be used in the cosmetic composition, and particularly, it is preferable to be used in the cosmetic composition because it has skin improvement, skin inflammation relief, and antioxidant effect.
본 발명의 화장료 조성물에서, 상기 항균 및 항진균용 화장료 조성물의 함량은 0.001 내지 10 중량%로 함유될 수 있다. In the cosmetic composition of the present invention, the content of the antimicrobial and antifungal cosmetic composition may be contained in 0.001 to 10% by weight.
본 발명의 화장료 조성물은 유효성분으로 상기 정향, 히비스커스, 및 코코넛 추출물 이외에 화장료 조성물에 통상적으로 이용되는 성분들이 포함되며, 예컨대 항산화제, 안정화제, 용해화제, 비타민, 안료 및 향료와 같은 통상적인 보조제, 그리고 담체를 포함한다.The cosmetic composition of the present invention contains ingredients commonly used in cosmetic compositions in addition to the cloves, hibiscus, and coconut extract as an active ingredient, and for example, conventional adjuvants such as antioxidants, stabilizers, solubilizers, vitamins, pigments and flavorings. And carriers.
본 발명의 화장료 조성물은 당업계에서 통상적으로 제조되는 어떠한 제형으로도 제조될 수 있으며, 예를 들어, 용액, 현탁액, 유탁액, 페이스트, 겔, 크림, 로션, 파우더, 비누, 계면활성제-함유 클렌징, 오일, 분말 파운데이션, 유탁액 파운데이션, 왁스 파운데이션 및 스프레이 등으로 제형화될 수 있으나, 이에 한정되는 것은 아니다. 보다 상세하게는, 유연 화장수(스킨), 영양 화장수(밀크로션), 영양 크림, 맛사지 크림, 에센스, 아이크림, 클렌징 크림, 클렌징 포옴, 클렌징 워터, 팩, 스프레이 또는 파우더의 제형으로 제조될 수 있다.Cosmetic compositions of the present invention may be prepared in any formulation conventionally prepared in the art and include, for example, solutions, suspensions, emulsions, pastes, gels, creams, lotions, powders, soaps, surfactant-containing cleansing , Oils, powder foundations, emulsion foundations, wax foundations and sprays, and the like, but are not limited thereto. More specifically, it may be prepared in the form of a flexible lotion (skin), nutrition lotion (milk lotion), nutrition cream, massage cream, essence, eye cream, cleansing cream, cleansing foam, cleansing water, pack, spray or powder .
본 발명의 제형이 페이스트, 크림 또는 겔인 경우에는 담체 성분으로서 동물성 유, 식물성 유, 왁스, 파라핀, 전분, 트라가칸타, 셀룰로오스 유도체, 폴리에틸렌 글리콜, 실리콘, 벤토나이트, 실리카, 탈크 또는 산화아연 등이 이용될 수 있다.When the formulation of the present invention is a paste, cream or gel, animal oil, vegetable oil, wax, paraffin, starch, tragacanta, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide are used as carrier components. Can be.
본 발명의 제형이 파우더 또는 스프레이인 경우에는 담체 성분으로서 락토스, 탈크, 실리카, 알루미늄 히드록시드, 칼슘 실리케이트 또는 폴리아미드 파우더가 이용될 수 있고, 특히 스프레이인 경우에는 추가적으로 클로로플루오로히드로카본, 프로판/부탄 또는 디메틸 에테르와 같은 추진체를 포함할 수 있다.When the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used, in particular in the case of a spray, additionally chlorofluorohydrocarbon, propane Propellant such as butane or dimethyl ether.
본 발명의 제형이 용액 또는 유탁액인 경우에는 담체 성분으로서 용매, 용해화제 또는 유탁화제가 이용되고, 예컨대 물, 에탄올, 이소프로판올, 에틸 카보네이트, 에틸 아세테이트, 벤질 알코올, 벤질 벤조에이트, 프로필렌글리콜, 1,3-부틸글리콜 오일, 글리세롤 지방족 에스테르, 폴리에틸렌 글리콜 또는 소르비탄의 지방산 에스테르가 있다.When the formulation of the present invention is a solution or emulsion, a solvent, solubilizer or emulsifier is used as the carrier component, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1 Fatty acid esters of, 3-butylglycol oil, glycerol aliphatic ester, polyethylene glycol or sorbitan.
본 발명의 제형이 현탁액인 경우에는 담체 성분으로서 물, 에탄올 또는 프로필렌 글리콜과 같은 액상의 희석제, 에톡실화 이소스테아릴 알코올, 폴리옥시에틸렌 소르비톨 에스테르 및 폴리옥시에틸렌 소르비탄 에스테르와 같은 현탁제, 미소 결정성 셀룰로오스, 알루미늄 메타히드록시드, 벤토나이트, 아가 또는 트라가칸타 등이 이용될 수 있다.When the formulation of the present invention is a suspension, liquid carrier diluents such as water, ethanol or propylene glycol, suspending agents such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol esters and polyoxyethylene sorbitan esters, and microcrystals are used as carrier components. Soluble cellulose, aluminum metahydroxy, bentonite, agar or tragacanta and the like can be used.
본 발명의 제형이 계면-활성제 함유 클렌징인 경우에는 담체 성분으로서 지방족 알코올 설페이트, 지방족 알코올 에테르 설페이트, 설포숙신산 모노에스테르, 이세티오네이트, 이미다졸리늄 유도체, 메틸타우레이트, 사르코시네이트, 지방산 아미드 에테르 설페이트, 알킬아미도베타인, 지방족 알코올, 지방산 글리세리드, 지방산 디에탄올아미드, 식물성 유, 라놀린 유도체 또는 에톡실화 글리세롤 지방산 에스테르 등이 이용될 수 있다.When the formulation of the present invention is a surfactant-containing cleansing, the carrier component is an aliphatic alcohol sulfate, an aliphatic alcohol ether sulfate, a sulfosuccinic acid monoester, an isethionate, an imidazolinium derivative, a methyltaurate, a sarcosinate, a fatty acid amide. Ether sulfates, alkylamidobetaines, aliphatic alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, lanolin derivatives or ethoxylated glycerol fatty acid esters and the like can be used.
본 발명의 화장품의 구체예로서는 여성청결제, 손세정제, 세안크림, 세안폼, 클렌징크림, 클렌징밀크, 클렌징로션, 마사지크림, 콜드크림, 모이스처크림, 유액, 화장수, 팩, 에프터세이빙크림, 썬텐방지크림, 썬텐용 오일, 비누, 보디샴푸, 헤어샴푸, 헤어린스, 헤어트리트먼트, 양모료, 육모료, 헤어크림, 헤어리퀴드, 세트로션, 헤어스프레이, 헤어브리지, 컬러린스, 컬러스프레이, 퍼머넌트웨이브액, 프레스파우더, 루스파우더, 아이섀도, 핸드크림, 및 립스틱 등을 들 수 있다.Specific examples of the cosmetics of the present invention include a female cleanser, hand cleanser, cleansing cream, cleansing foam, cleansing cream, cleansing milk, cleansing lotion, massage cream, cold cream, moisturizing cream, latex, lotion, pack, aftershave cream, sunburn prevention cream , Suntan oil, soap, body shampoo, hair shampoo, hair rinse, hair treatment, wool, hair restorer, hair cream, hair liquid, set lotion, hair spray, hair bridge, coloring, color spray, permanent wave liquid , Press powder, loose powder, eye shadow, hand cream, lipstick, and the like.
본 발명의 또 다른 양태에 따르면, 본 발명은 정향, 히비스커스, 및 코코넛 추출물을 유효성분으로 포함하는 항균 및 항진균용 식품 조성물 또는 사료 보존 조성물을 제공한다.According to another aspect of the present invention, the present invention provides a food composition or feed preservation composition for antibacterial and antifungal, comprising clove, hibiscus, and coconut extract as an active ingredient.
식품이나 사료의 보존제는 식품이나 사료의 변질, 부패, 변색 및 화학변화를 방지하기 위해 사용되는 첨가물로서 살균제, 산화방지제가 이에 포함되며, 세균, 곰팡이, 효모 등 미생물의 증식을 억제하여 식품, 사료, 화장품, 의약품 등에서 부패 미생물의 발육저지 또는 살균작용을 하는 등의 기능성 항균제도 포함된다. 이러한 식품이나 사료의 방부제의 이상적인 조건으로는 독성이 없어야 하며, 미량으로도 효과가 있어야 한다. 본 발명의 정향, 히비스커스, 및 코코넛 추출물을 유효성분으로 포함하는 항균 및 항진균용 식품 조성물 또는 사료 보존 조성물은 유해 미생물에 직접 영향을 미치는 것을 통해 탁월한 항균 및 항진균 활성을 나타내므로 상기의 식품이나 사료의 보존제, 화장품 보존제 및 의약품 보존제 등에 폭넓게 사용될 수 있다.Food and feed preservatives are additives used to prevent the deterioration, decay, discoloration and chemical change of food or feed, including fungicides and antioxidants, and inhibit the growth of microorganisms such as bacteria, fungi and yeasts. Functional antibacterial agents, such as inhibiting the development or sterilization of the decaying microorganisms in cosmetics, cosmetics, and the like are included. Ideal conditions for preservatives of these foods or feeds should be nontoxic and effective in trace amounts. The antimicrobial and antifungal food composition or feed preservation composition comprising clove, hibiscus, and coconut extract of the present invention as an active ingredient exhibits excellent antimicrobial and antifungal activity through direct influence on harmful microorganisms. It can be widely used for preservatives, cosmetic preservatives and pharmaceutical preservatives.
본 발명의 또 다른 양태에 따르면, 본 발명은 상기 정향, 히비스커스, 및 코코넛 추출물을 유효성분으로 포함하는 항균 및 항진균 조성물을 치료를 필요로 하는 개체에 투여하여 병원성 세균을 사멸시키는 단계를 포함하는, 인간을 제외한 동물의 세균 또는 진균성 감염 질환을 치료하는 방법을 제공한다.According to another aspect of the invention, the present invention comprises the step of killing pathogenic bacteria by administering to the subject in need of the antimicrobial and antifungal composition comprising the clove, hibiscus, and coconut extract as an active ingredient, Provided are methods for treating bacterial or fungal infectious diseases in animals other than humans.
본 발명에 따른 상기 인간을 제외한 동물의 세균 또는 진균성 감염 질환을 치료하는 방법은 비록 인간을 제외한 동물을 치료하는 방법이나, 인간에 있어 이러한 치료 방법이 효과가 없음을 의미하는 것은 아니다. 또한, 인간의 경우 있어서 본 발명에 따른 치료용 조성물의 투여에 의해 증상이 호전될 수 있는 세균 또는 진균 감염에 의한 질환을 갖는 것을 고려할 때, 인간의 치료에 있어서도 충분히 사용될 수 있다.Although the method for treating bacterial or fungal infectious diseases of animals other than humans according to the present invention does not mean that the method of treating animals except humans is not effective in humans. In addition, in the case of humans, when considering the disease caused by bacterial or fungal infection, the symptoms of which can be improved by administration of the therapeutic composition according to the present invention can be sufficiently used in the treatment of humans.
본 발명에서 용어 "인간을 제외한 동물"은 본 발명에 따른 치료용 조성물의 투여에 의해 증상이 호전될 수 있는 세균 또는 진균의 감염에 의해 유발되는 질환을 갖는 인간만을 제외한 말, 양, 돼지, 염소, 낙타, 영양, 개 등의 동물을 의미한다. 본 발명에 따른 치료용 조성물을 인간을 제외한 동물에게 투여함으로써, 세균성 또는 진균성 유해 미생물 감염에 의한 질환을 효과적으로 예방 및 치료할 수 있다.In the present invention, the term "animal other than human" refers to horses, sheep, pigs and goats, except for humans having diseases caused by infection of bacteria or fungi whose symptoms may be improved by administration of the therapeutic composition according to the present invention. Means animals such as camels, antelopes and dogs. By administering the therapeutic composition according to the present invention to animals other than humans, diseases caused by bacterial or fungal harmful microbial infections can be effectively prevented and treated.
본 발명에서 용어 "투여"는 어떠한 적절한 방법으로 동물에게 소정의 물질을 도입하는 것을 의미하며, 본 발명에 따른 치료용 조성물의 투여 경로는 목적 조직에 도달할 수 있는 한 어떠한 일반적인 경로를 통하여 경구 또는 비경구 투여될 수 있다. 또한, 본 발명에 따른 치료용 조성물은 유효성분이 표적 세포로 이동할 수 있는 임의의 장치에 의해 투여될 수 있다.The term "administration" in the present invention means introducing a predetermined substance into the animal by any suitable method, and the route of administration of the therapeutic composition according to the present invention is oral or via any general route as long as it can reach the target tissue. Parenteral administration. In addition, the therapeutic composition according to the present invention can be administered by any device that the active ingredient can move to the target cell.
본 발명에 따른 치료용 조성물의 바람직한 투여량은 치료 대상 동물의 상태 및 체중, 질병의 정도, 약물형태, 투여경로 및 기간에 따라 다르지만, 당업자에 의해 적절하게 선택될 수 있다. 그러나, 바람직한 효과를 위해서, 본 발명의 약학적 조성물은 0.01 내지 10 ㎎/㎏이고, 바람직하게는 0.1 내지 1 ㎎/㎏ 이며, 하루 1회 내지 3회 투여할 수 있다.Preferred dosages for the therapeutic compositions according to the invention vary depending on the condition and weight of the animal to be treated, the extent of the disease, the form of the drug, the route of administration and the duration, and may be appropriately selected by those skilled in the art. However, for the desired effect, the pharmaceutical composition of the present invention is 0.01 to 10 mg / kg, preferably 0.1 to 1 mg / kg, and may be administered once to three times a day.
본 발명의 조성물은 그 자체적으로 항균 및 항진균 효과가 있으므로 항균 및 항진균 조성물로 바로 이용될 수 있고, 항균 및 항진균 효과가 있는 천연 추출물들을 함유하는 다른 성분과 혼합하여 사용하는 형태로도 이용이 가능하여 기존의 항균 또는 항진균 효과를 좀 더 높여줄 수 있을 것으로 기대된다.Since the composition of the present invention has an antibacterial and antifungal effect on its own, it can be directly used as an antibacterial and antifungal composition, and can be used in the form of mixing with other ingredients containing natural extracts having antibacterial and antifungal effects. It is expected that the antimicrobial or antifungal effect can be enhanced.
본 발명의 특징 및 이점을 요약하면 다음과 같다:The features and advantages of the present invention are summarized as follows:
(1) 본 발명은 정향, 히비스커스, 및 코코넛 추출물을 유효성분으로 포함하는 항균 및 항진균 조성물을 제공한다.(1) The present invention provides an antimicrobial and antifungal composition comprising clove, hibiscus, and coconut extract as an active ingredient.
(2) 본 발명의 정향, 히비스커스, 및 코코넛 추출물을 유효성분으로 포함하는 항균 및 항진균 조성물은 다양한 균들에 대해 폭넓은 항균 및 항진균 스펙트럼을 나타내는 동시에, 인간 진피 섬유 아세포에서 MMP-1의 발현을 억제하고, 피부 주름의 발생을 억제하며, 손상된 피부의 염증반응을 개선하는 효능을 나타낸다. (2) The antimicrobial and antifungal composition comprising the clove, hibiscus, and coconut extract of the present invention as an active ingredient exhibits a broad antimicrobial and antifungal spectrum against various bacteria, while inhibiting the expression of MMP-1 in human dermal fibroblasts. Inhibits the occurrence of skin wrinkles, and improves the inflammatory response of damaged skin.
(3) 또한, 본 발명의 조성물은 화장료 조성물, 식품 또는 사료 보존용 첨가제 조성물 등의 다양한 분야에서 항균, 항진균 및 방부 목적으로 유용하게 이용 가능하다.(3) The composition of the present invention can also be usefully used for antibacterial, antifungal and antiseptic purposes in various fields such as cosmetic compositions, food or feed preservative additive compositions.
도 1은 본 발명의 정향, 히비스커스, 및 코코넛의 복합추출물이 갖는 MMP-1 발현 억제 효과를 나타내는 그래프이다.
도 2는 본 발명의 정향, 히비스커스, 및 코코넛의 복합추출물이 갖는 IL-6 억제 효과를 나타내는 그래프이다.
도 3은 본 발명의 정향, 히비스커스, 및 코코넛의 복합추출물이 갖는 항산화 효과를 나타내는 그래프이다.1 is a graph showing the inhibitory effect of MMP-1 expression of the complex extract of clove, hibiscus, and coconut of the present invention.
Figure 2 is a graph showing the IL-6 inhibitory effect of the complex extract of clove, hibiscus, and coconut of the present invention.
Figure 3 is a graph showing the antioxidant effect of the complex extract of clove, hibiscus, and coconut of the present invention.
이하, 실시예를 통하여 본 발명을 더욱 상세히 설명하기로 한다. 이들 실시예는 단지 본 발명을 예시하기 위한 것이므로, 본 발명의 범위가 이들 실시예에 의해 제한되는 것으로 해석되지는 않는다. 아래의 실시예에서 사용되는 %는 별도의 언급이 없는 경우, 고체/고체는 중량/중량%, 고체/액체는 중량/부피%, 그리고 액체/액체는 부피/부피%를 각각 나타낸다.Hereinafter, the present invention will be described in more detail with reference to Examples. Since these examples are only for illustrating the present invention, the scope of the present invention is not to be construed as being limited by these examples. The percentages used in the examples below refer to weight / weight percent solids / solids, weight / volume percent solids / liquids, and volume / volume percent liquids / liquids, unless otherwise noted.
실시예 1. 식물 추출물의 제조Example 1 Preparation of Plant Extract
1-1. 정향 추출물의 제조1-1. Preparation of Clove Extract
건조 중량 100g의 분쇄한 정향을 70%(V/V) 에탄올 수용액으로 12시간 가온 환류추출하고 냉침한 후, 1.2㎛ 투과사이즈를 갖는 여과지(Glassfiber, sartorius, 미국)로 여과하였다. 이 여과된 추출액을 50℃ 이하에서 감압농축한 후 감압건조기(Eyela Rotary evaporator, Tokyo Rikakika Co.,LTD, 일본)를 이용해 완전히 건조시켜 정향 추출물 12g을 수득하였다.The dried cloves, weighing 100 g of dry weight, were refluxed for 12 hours with a 70% (V / V) ethanol aqueous solution and cooled down, and then filtered through a filter paper (Glassfiber, sartorius, USA) having a permeation size of 1.2 μm. The filtered extract was concentrated under reduced pressure at 50 ° C. or lower and completely dried using a reduced pressure dryer (Eyela Rotary evaporator, Tokyo Rikakika Co., Ltd., Japan) to obtain 12 g of clove extract.
1-2. 히비스커스 추출물의 제조1-2. Preparation of Hibiscus Extract
건조 중량 100g의 분쇄한 히비스커스를 70%(V/V) 에탄올 수용액으로 12시간 가온 환류추출하고 냉침한 후, 1.2㎛ 투과사이즈를 갖는 여과지(Glassfiber, sartorius, 미국)로 여과하였다. 이 여과된 추출액을 50℃ 이하에서 감압농축한 후 감압건조기(Eyela Rotary evaporator, Tokyo Rikakika Co.,LTD, 일본)를 이용해 완전히 건조시켜 히비스커스 추출물 31g을 수득하였다.The dried hibiscus with a dry weight of 100 g was heated and refluxed for 12 hours with an aqueous 70% (V / V) ethanol solution, cooled and then filtered through a filter paper (Glassfiber, sartorius, USA) having a permeation size of 1.2 μm. The filtered extract was concentrated under reduced pressure at 50 ° C. or lower and completely dried using a reduced pressure dryer (Eyela Rotary evaporator, Tokyo Rikakika Co., Ltd., Japan) to obtain 31 g of a hibiscus extract.
1-3. 코코넛 추출물의 제조1-3. Preparation of Coconut Extract
건조 중량 100g의 분쇄한 코코넛을 70%(V/V) 에탄올 수용액으로 12시간 가온 환류추출하고 냉침한 후, 1.2㎛ 투과사이즈를 갖는 여과지(Glassfiber, sartorius, 미국)로 여과하였다. 이 여과된 추출액을 50℃ 이하에서 감압농축한 후 감압건조기(Eyela Rotary evaporator, Tokyo Rikakika Co.,LTD, 일본)를 이용해 완전히 건조시켜 코코넛 추출물 12g을 수득하였다.100 g of dry weight of crushed coconut was extracted with warm reflux for 12 hours with 70% (V / V) ethanol aqueous solution, and then cooled and filtered through a filter paper (Glassfiber, sartorius, USA) having a permeation size of 1.2 μm. The filtered extract was concentrated under reduced pressure at 50 ° C. or lower and completely dried using a reduced pressure dryer (Eyela Rotary evaporator, Tokyo Rikakika Co., Ltd., Japan) to obtain 12 g of coconut extract.
실시예 2. 단일 화합물에 대한 항균 활성 측정Example 2. Determination of Antimicrobial Activity against Single Compounds
2-1. 균주 및 배양조건2-1. Strains and Culture Conditions
상기 추출물들의 항균 효과를 측정하기 위하여 디스크 확산법 실험을 수행하여 항균 활성을 조사하였다. 항균 효과를 측정하기 위하여 사용한 균주, 배지, 및 배양온도는 아래 [표 1]에 나타내었다. In order to determine the antimicrobial effect of the extracts were carried out disk diffusion experiment to investigate the antimicrobial activity. The strain, medium, and culture temperature used to measure the antimicrobial effect are shown in Table 1 below.
[표 1]TABLE 1
대장균(E coli KCTC2571), 슈도모나스 에루지노사(Pseudomonas aeruginosa KCTC2513), 스타필로코커스 아우레우스(Staphylococcus aureus KCTC3881) 및 바실러스 서브틸리스(Bacillus subtilis KCTC1021)의 균주와, 진균류인 칸디다 알비칸스(Candida albicans KCTC7965), 아스퍼질러스 나이거(Aspergillus niger KCTC6317)를 사용하였다(KCTC 한국생명공학연구원 생물자원센터).E. coli KCTC2571, Pseudomonas aeruginosa KCTC2513, Staphylococcus aureus KCTC3881 and Bacillus subtilis KCTC1021, and the fungus albicans albicans KCTC7965) and Aspergillus niger (Aspergillus niger KCTC6317) were used (KCTC Bioresource Center, Korea Research Institute of Bioscience and Biotechnology).
2-2. 디스크 확산법에 의한 항균력 검증2-2. Antibacterial activity verification by disk diffusion method
각각의 균주는 상기 표 1의 영양배지(Nutrient Broth) 용액을 제조하여 121℃에서 15분간 멸균한 후 각각의 배양된 균주 1 백금이를 영양배지 용액 (10㎖)에 접종하고, 진탕배양기(shaking incubator)에서 24시간 배양하여 항균실험을 위한 균주를 준비하였다.Each strain prepared the Nutrient Broth solution of Table 1 and sterilized at 121 ° C. for 15 minutes, and then inoculated each
미생물 균주의 성장을 억제하는 항생물질의 최소억제농도를 측정하기 위해, 멸균된 배지에 배양된 균주를 세균은 1 X 106 cfu/㎖, 진균은 1 X 105 cfu/㎖의 농도로 각각을 접종하였다. 상기 실시예 1에서 제조된 3종의 단일 추출물은 에탄올에 2% 농도로 용해시켜 멸균한 페이퍼 디스크(직경 8mm)에 25㎕씩 두 번 점적한 후 충분히 건조시켰다. 추출물이 완전히 건조된 다음 평판 배지에 치상하여 28~37℃로 유지하여, 각각 1, 2, 3일(A. niger의 경우 7일까지 관찰) 후, 페이퍼 디스크 주변의 투명도(clear zone)를 측정하여 항균 활성을 조사하였다. To measure the minimum inhibitory concentration of the antibiotic which inhibits growth of the microbial strain, bacteria of the strains cultured in a sterile culture medium is 1 X 10 6 cfu / ㎖, fungi, each at a concentration of 1 X 10 5 cfu / ㎖ Inoculation. Three single extracts prepared in Example 1 were dissolved in ethanol at a concentration of 2%, dropped twice at 25 μl on sterile paper discs (diameter 8 mm) and dried sufficiently. After the extract is completely dried, it is placed on a plate medium and maintained at 28-37 ° C. After 1, 2 and 3 days (observe up to 7 days for A. niger), the clear zone around the paper disc is measured. The antimicrobial activity was investigated.
상기 추출물들의 항균력을 비교 검증하기 위해, 시중에서 상용되는 메틸파라벤(Methyl Paraben)을 비교예 1로, 자몽 종자 추출물을 비교예 2로 사용하여 비교 실험하였다. In order to compare and verify the antimicrobial activity of the extracts, commercially available commercial commercial methyl paraben (Methyl Paraben) in Comparative Example 1, Comparative Example 2 using grapefruit seed extract.
상기 정향, 히비스커스, 및 코코넛의 단일 추출물의 디스크 확산법 실험에 따른 항균 활성을 아래 표 2에 나타내었다. The antimicrobial activity of the clove, hibiscus, and disc diffusion experiments of a single extract of coconut is shown in Table 2 below.
[표 2]TABLE 2
그 결과, 정향 추출물의 경우에는 자몽 종자 추출물(비교예 2)과 비교하여 유사한 정도의 항균 활성을 갖는 것으로 나타났으나, 히비스커스 또는 코코넛 추출물은 메틸파라벤(비교예 1)과 비교하여 비교 열위의 항균 활성을 갖는 것이 관찰되었다. As a result, the extract of clove was found to have a similar degree of antimicrobial activity as compared to the grapefruit seed extract (Comparative Example 2), but the hibiscus or coconut extract was compared to methylparaben (Comparative Example 1). It was observed to have activity.
실시예 3. 정향, 히비스커스, 및 코코넛의 복합 추출물이 갖는 항균 활성 측정Example 3 Determination of Antimicrobial Activity of Complex Extracts of Clove, Hibiscus, and Coconut
3-1. 단일 추출물의 MIC 평가3-1. MIC Evaluation of Single Extracts
정향, 히비스커스, 및 코코넛의 단일 추출물에 대한 항균 활성을 측정하기 위하여 실시예 1에서 사용한 유해 미생물을 대상으로 미생물 생장 최소억제농도(MIC)를 평가하였다.The microbial growth minimum inhibitory concentration (MIC) was evaluated for the harmful microorganisms used in Example 1 to determine the antimicrobial activity of a single extract of clove, hibiscus, and coconut.
MIC 측정은 고형 한천 배지에서 각 세균과 진균을 배양한 후 1×105~6 CFU/mL에 준하여 5개의 집락을 얻고 배지에 희석하여 접종한 후 37℃에서 2~3시간 동안 유지하였다. MIC measurement was obtained by incubating each bacteria and fungi in a solid agar medium, 5 colonies were obtained according to 1 × 10 5-6 CFU / mL, diluted in the medium and inoculated and maintained at 37 ° C. for 2-3 hours.
본 실험에 사용한 식물추출물은 2.0mg/mL 농도로 적정하여 각각 2배 계열로 희석하였다. 각 세균과 화합물이 함유된 배지를 96 well plate에 옮기고 37℃에서 16~24시간 동안 배양한 후, 600nm에서 Multilabel Counter(1420, Perkin Elmer)로 측정하여 단일추출물들의 최소저해농도를 구하고, 이를 하기 [표 3]에 나타내었다.Plant extracts used in this experiment were titrated at a concentration of 2.0mg / mL and diluted in 2-fold series. Transfer the medium containing each bacterium and compound to a 96 well plate and incubate at 37 ° C. for 16-24 hours, and measure the minimum inhibitory concentration of single extracts by measuring it at 600 nm using the Multilabel Counter (1420, Perkin Elmer). It is shown in [Table 3].
[표 3]TABLE 3
그 결과, 정향, 히비스커스 및 코코넛 추출물 모두 메틸파라벤(비교예 1) 또는 자몽 종자 추출물(비교예 2)과 대비하여 비교 열위의 항균 활성을 갖는 것이 관찰되었다. As a result, it was observed that the clove, hibiscus and coconut extracts all had comparative inferior antimicrobial activity as compared to methylparaben (Comparative Example 1) or grapefruit seed extract (Comparative Example 2).
3-2. 정향과 히비스커스 복합 추출물의 MIC 평가3-2. MIC Evaluation of Clove and Hibiscus Complex Extract
정향과 히비스커스 추출물 사이의 상호 영향을 확인하기 위하여 이들 복합 추출물의 미생물 생장 최소억제농도(MIC)를 평가하였다.The microbial growth inhibitory concentration (MIC) of these complex extracts was evaluated to confirm the mutual effect between the clove and hibiscus extract.
항균 시너지를 확인하기 위한 실험 방법은 표준화되어 있지 않아, 항균물질을 조합하여 항균 시너지를 확인하는 방법인 체크보드 어세이(checkboard assay)를 응용하여 사용하였다. 항균력 측정실험은 6행×6열로 설정하였고, 96 웰-플레이트에 혼합효과를 측정할 정향과 히비스커스 추출물을 각각 MIC 농도에서 연속 2배 희석하여 정향 추출물은 수평축으로 히비스커스 추출물은 수직축으로 각 웰에 연속적으로 희석하였다. 이후 37℃에서 16~24시간 동안 배양한 후, 복합 추출물의 상호적으로 억제한 가장 낮은 농도로서 정의하였으며, 두 화합물 사이의 상호작용은 하기 [수학식 1]의 FIC(fractional inhibitory concentration)로 나타내었다. Experimental method for checking the antimicrobial synergy is not standardized, it was used by applying a checkboard assay (checkboard assay) which is a method for identifying the antimicrobial synergy by combining antimicrobial substances. The antimicrobial activity was set to 6 rows x 6 columns, and the clove and hibiscus extracts, which were used to measure the mixing effect on 96 well plates, were diluted two-fold at MIC concentrations, and the clove extracts were horizontal and the hibiscus extracts were vertical. Diluted with. After incubating for 16-24 hours at 37 ℃, it was defined as the lowest mutually inhibited concentration of the complex extract, the interaction between the two compounds is represented by the FIC (fractional inhibitory concentration) of the following [Equation 1] It was.
FICI(FIC index)는 하기 [식 1] 및 [식 2]에 따라 계산되었다.FICI (FIC index) was calculated according to the following [Formula 1] and [Formula 2].
[수학식 1][Equation 1]
[수학식 2][Equation 2]
이때, FIC 지수가 0.5보다 작거나 같으면 상승효과(synergistic effect), 0.5 내지 1에 해당하면 부가 효과(additive effect), 1 내지 4에 해당하면 무관함(indifferent), 4 이상이면 상쇄 효과(antagonistic effect)가 있는 것으로 판단하였고, 측정 결과를 하기 [표 4] 내지 [표 9]에 나타내었다.In this case, if the FIC index is less than or equal to 0.5, a synergistic effect, an additive effect for 0.5 to 1, an independent effect for 1 to 4, and an antagonistic effect for 4 or more ), And the measurement results are shown in the following [Table 4] to [Table 9].
[표 4] C. albicans Table 4 C. albicans
[표 5] A. niger Table 5 A. niger
[표 6] E. coli Table 6 E. coli
[표 7] P. aeruginosa Table 7 P. aeruginosa
[표 8] B. subtilis Table 8 B. subtilis
[표 9] S. aurues Table 9 S. aurues
실험 결과, 정향 추출물과 히비스커스 추출물이 1:8의 비율로 혼합된 조성물이 모든 유해 미생물에 대해 상승 또는 부가된 항균 및 항진균 효과를 갖는 것을 확인할 수 있었고, 하기 [표 10]의 비교표를 참조하면, 정향 추출물과 히비스커스 추출물이 1:8의 비율로 혼합된 조성물은 비교예의 메틸파라벤 또는 자몽종자 추출물과 유사한 수준의 항균활성을 나타내어, 가장 우수한 시너지 효과가 있는 것을 확인할 수 있었다. As a result, it was confirmed that the composition of the clove extract and hibiscus extract in a ratio of 1: 8 has a synergistic or added antimicrobial and antifungal effect on all harmful microorganisms, and referring to the comparison table of the following [Table 10], The composition of the clove extract and hibiscus extract in a ratio of 1: 8 showed an antimicrobial activity similar to that of the methylparaben or grapefruit seed extract of the comparative example, it was confirmed that the most synergistic effect.
[표 10]TABLE 10
3-3. 3-3. 정향cloves 및 And 히비스커스와Hibiscus and 코코넛의 복합 추출물에 대한 For complex extract of coconut MICMIC 평가 evaluation
정향 및 히비스커스 추출물과 코코넛 추출물 사이의 상호 영향을 확인하기 위하여 이들 복합 추출물의 비율에 따른 미생물 생장 최소억제농도(MIC)를 측정하였다. 실험은 항균 및 항진균 효과가 가장 좋은 정향 및 히비스커스의 혼합 비율(1:8)을 고정시키고, 이를 코코넛 추출물과 혼합하면서 상기 실시예 3-3의 방법과 동일한 방법으로 평가하였고, 측정 결과를 하기 [표 11]에 정리하였다.In order to confirm the mutual influence between the clove and hibiscus extract and coconut extract, the microbial growth minimum inhibition concentration (MIC) was measured according to the ratio of these complex extracts. The experiment was evaluated in the same manner as in Example 3-3 while fixing the mixing ratio (1: 8) of the clove and hibiscus best antibacterial and antifungal effect, and mixed with the coconut extract, the measurement results [ Table 11 summarized.
[표 11] TABLE 11
실험 결과, 정향 및 히비스커스의 복합 추출물(1:8)과 코코넛 추출물이 1:1의 비율로 혼합된 조성물이 모든 유해 미생물에 대해 상승 또는 부가된 항균 및 항진균 효과를 갖는 것을 확인할 수 있었다. As a result, it was confirmed that the composition in which the complex extract of clove and hibiscus (1: 8) and the coconut extract were mixed at a ratio of 1: 1 has an antibacterial and antifungal effect on all harmful microorganisms.
하기 [표 12]의 비교표를 참조하면, 정향, 히비스커스, 및 코코넛 추출물이 1:8:9의 비율로 혼합된 복합 추출물은 비교예 1의 메틸파라벤과 대비하여 매우 유사한 수준의 항균 활성을 나타내어, 가장 우수한 시너지 효과가 있는 것을 확인할 수 있었다. Referring to the comparative table of the following [Table 12], the complex extract of clove, hibiscus, and coconut extract in a ratio of 1: 8: 9 shows a very similar level of antimicrobial activity compared to methylparaben of Comparative Example 1, It was confirmed that the best synergy effect.
[표 12]TABLE 12
또한, 정향, 히비스커스, 및 코코넛 추출물이 1:8:9의 비율로 혼합된 복합 추출물은 각각의 단일 추출물의 평균값 대비 97.6%-99.3%, 시험예 4의 정향 및 히비스커스의 2종 복합추출물 대비 50%-75% 감소된 유해 미생물 생장 최소억제농도가 측정되어, 가장 우수한 시너지 효과가 있는 것을 확인할 수 있었다. In addition, the composite extract of clove, hibiscus, and coconut extract in a ratio of 1: 8: 9 is 97.6% -99.3% of the average value of each single extract, 50 of the two complex extracts of clove and hibiscus of Test Example 4 % -75% reduced harmful microbial growth minimum concentration was measured, it was confirmed that the best synergistic effect.
실시예 4. 정향, 히비스커스, 및 코코넛 추출물의 주름개선 효과 측정Example 4 Wrinkle Improvement Effect Measurement of Clove, Hibiscus, and Coconut Extract
건강한 젊은 남성 지원자(SD 생명공학, 서울, 한국)를 선발하고, 피부 생검하여 정상 인간 진피 섬유아세포(NHDF)들을 획득하였다. 세포들은 100㎜조직 배양 플레이트에서 플레이팅 되었고, 5% CO2를 함유하는 습기 환경, 37℃에서 10%의 열-비활성화된 FBS 및 1%의 페니실린-스트렙토마이신으로 보충된 DMEM 배지에서 배양되었다. 모든 실험은 오직 계대 6 내지 10 사이의 세포들을 사용하여 수행하였다.Healthy young male volunteers (SD Biotechnology, Seoul, Korea) were selected and skin biopsied to obtain normal human dermal fibroblasts (NHDFs). Cells were plated in 100 mm tissue culture plates and cultured in DMEM medium supplemented with 10% heat-inactivated FBS and 1% penicillin-streptomycin at 37 ° C. in a humid environment containing 5% CO 2 . All experiments were performed using only cells between passages 6-10.
상기 정상 인간 진피 섬유아세포들은 40㎜ 조직 배양 플레이트들(1.2×105 세포들)에 시딩하였다. 정상 인간 진피 섬유아세포들이 80% 융합에 도달할 때, 그들을 인산 완충 식염수(PBS)로 두 차례 씻어내었고, 신선한 세럼-프리 배지 1980㎕와 샘플 20㎕는 각 웰에 첨가된 후, 정상 인간 진피 섬유아세포들은 따뜻한 PBS로 세 차례 세척하였다.The normal human dermal fibroblasts were seeded in 40 mm tissue culture plates (1.2 × 10 5 cells). When normal human dermal fibroblasts reached 80% fusion, they were washed twice with phosphate buffered saline (PBS) and 1980 μl of fresh serum-free medium and 20 μl of sample were added to each well and then normal human dermis Fibroblasts were washed three times with warm PBS.
MMP-1(매트릭스 메탈로프로테이나제) 활성은 UVB 조사 후 72시간 채취된 상층액에서 평가되었다. ELISA 리더기를 이용하여 채취된 상층액은 450nm에서 측정 되었다.MMP-1 (matrix metalloproteinase) activity was evaluated in supernatants taken 72 hours after UVB irradiation. Supernatants collected using an ELISA reader were measured at 450 nm.
그 결과, 본 발명의 정향, 히비스커스, 및 코코넛의 복합추출물(시험예 5: 1:8:9 중량비)이 처리된 피부세포에서 농도의존적으로 세포생존율이 증가하고, MMP-1 활성이 억제되는 것을 확인할 수 있었다(도 1). As a result, in the skin cells treated with the complex extract of the clove, hibiscus, and coconut (Test Example 5: 1: 8: 9 weight ratio) of the present invention, the cell survival rate was increased in a concentration-dependent manner, and MMP-1 activity was inhibited. It could be confirmed (Fig. 1).
실시예 5. 정향, 히비스커스, 및 코코넛 추출물의 항염증 효과 측정Example 5 Determination of Anti-inflammatory Effects of Clove, Hibiscus, and Coconut Extracts
HaCaT cells (피부 표피 각질 세포주) 세포들은 100㎜ 조직 배양 플레이트에서 플레이팅 되었고, 5% CO2를 함유하는 습기 환경, 37℃에서 10%의 열-비활성화된 FBS 및 1%의 페니실린-스트렙토마이신으로 보충된 DMEM 배지에서 배양되었다. HaCaT cells The cells were plated in 100 mm tissue culture plates, with a humid environment containing 5% CO 2 , 10% heat-inactivated FBS and 1% penicillin-streptomycin at 37 ° C. Cultured in supplemented DMEM medium.
정상 인간 표피 각질 세포 (HaCaT cells)들은 40㎜ 조직 배양 플레이트들(1.0×105 세포들)에 시딩하였다. 정상 인간 진피 섬유아세포들이 80% 융합에 도달할 때, 그들을 인산 완충 식염수(PBS)로 두 차례 씻어내었고, 신선한 세럼-프리 배지 1980㎕와 샘플 20㎕는 각 웰에 첨가된 후, HaCaT cells은 따뜻한 PBS로 세 차례 세척하였다.Normal human epidermal keratinocytes (HaCaT cells) were seeded in 40 mm tissue culture plates (1.0 × 10 5 cells). When normal human dermal fibroblasts reached 80% fusion, they were washed twice with phosphate buffered saline (PBS), 1980 μl of fresh serum-free medium and 20 μl of sample were added to each well, and then HaCaT cells Wash three times with warm PBS.
IL-6 단백질은 UVB 조사 후 24시간 채취된 상층액에서 평가되었다. ELISA 리더기를 이용하여 채취된 상층액을 450nm에서 측정하였다.IL-6 protein was evaluated in supernatants taken 24 hours after UVB irradiation. Supernatants collected using an ELISA reader were measured at 450 nm.
그 결과, 본 발명의 정향, 히비스커스, 및 코코넛의 복합추출물(시험예5: 1:8:9 중량비)이 처리된 피부세포에서 농도 의존적으로 세포 생존율이 증가하고, 염증매개 인자인 IL-6 단백질의 발현이 억제되는 것을 확인할 수 있었다(도 3).As a result, cell viability increased concentration-dependently in skin cells treated with the complex extract of the clove, hibiscus, and coconut (Test Example 5: 1: 8: 9 weight ratio) of the present invention, and the IL-6 protein, an inflammation mediating factor. It was confirmed that the expression of is suppressed (Fig. 3).
실시예 6. 정향, 히비스커스, 및 코코넛 추출물의 항산화 효과 검증Example 6. Antioxidant Effect of Clove, Hibiscus, and Coconut Extract
DPPH(1,1-diphenyl-2-picrylhydrazyl, Sigma, USA)의 환원력을 이용하는 상 등의 문헌(Sang S et al., J.Agric. Food Chem. 50(8), p2459-2463, 2002)에 기재된 방법에 따라, 본 발명에 따른 정향, 히비스커스, 및 코코넛의 복합추출물이 갖는 항산화 활성을 측정하였다.Sang S et al., J. Agric. Food Chem. 50 (8), p2459-2463, 2002, et al., Using a reducing power of DPPH (1,1-diphenyl-2-picrylhydrazyl, Sigma, USA). According to the described method, the antioxidant activity of the complex extract of clove, hibiscus, and coconut according to the present invention was measured.
메탄올에 녹인 0.2mM DPPH 용액 180ul와 증류수 20ul에 녹인 ‘정향, 히비스커스, 및 코코넛의 복합추출물’과 알부틴(양성대조군)(1, 10, 50, 100, 및 250㎍/㎖)을 각각 96-웰 마이크로플레이트에 넣고 37℃에서 30분간 배양한 후, 환원된 DPPH의 양을 분광 광도계(Precision microplate reader; Molecular Devices, VersaMax)를 이용하여 520nm에서 흡광도를 측정하여 시료를 가하지 않는 무첨가 대조군과 활성을 비교하였다. 상기 실험은 3회 반복하여 수행되었다. 96-well of 180 μl 0.2mM DPPH solution dissolved in methanol and 20 μl of distilled water, 'complex extract of clove, hibiscus, and coconut' and arbutin (positive control) (1, 10, 50, 100, and 250 μg / ml) After incubation at 37 ° C. for 30 minutes in a microplate, the amount of reduced DPPH was measured using a spectrophotometer (Precision microplate reader; Molecular Devices, VersaMax), and the absorbance was measured at 520 nm to compare the activity with no added control. It was. The experiment was performed three times.
그 결과, 정향, 히비스커스, 및 코코넛의 복합추출물(시험예5: 1:8:9 중량비)은 우수한 항산화 효과를 보이는 것을 관찰할 수 있었고, 이러한 항산화 효과의 수치들은 양성대조군인 알부틴과 비교하였을 때도 더 우수한 것으로 나타났다(도 3 참조).As a result, it was observed that the extract of clove, hibiscus, and coconut (Test Example 5: 1: 8: 9 weight ratio) showed an excellent antioxidant effect, even when compared with the arbutin positive control group. It was shown to be better (see FIG. 3).
이상으로 본 발명의 특정한 부분을 상세히 기술하였는 바, 당업계의 통상의 지식을 가진 자에게 있어서 이러한 구체적인 기술은 단지 바람직한 구현예일 뿐이며, 이에 본 발명의 범위가 제한되는 것이 아닌 점은 명백하다. 따라서, 본 발명의 실질적인 범위는 첨부된 청구항과 그의 등가물에 의하여 정의된다고 할 것이다.Having described the specific part of the present invention in detail, it is apparent to those skilled in the art that the specific technology is merely a preferred embodiment, and the scope of the present invention is not limited thereto. Thus, the substantial scope of the present invention will be defined by the appended claims and equivalents thereof.
Claims (10)
상기 정향, 히비스커스, 및 코코넛은 1:8:9의 중량비를 갖는 것을 특징으로 하는,
항균 및 항진균 조성물.Clove, hibiscus, and coconut extract as an active ingredient,
Wherein said clove, hibiscus, and coconut have a weight ratio of 1: 8: 9,
Antibacterial and antifungal composition.
상기 조성물은 칸디다 속 진균 및 아스퍼질러스 속 진균에 대해 항진균 활성을 갖는 것을 특징으로 하는 항균 및 항진균 조성물.The method of claim 1,
The composition is antibacterial and antifungal composition, characterized in that it has antifungal activity against fungi of the genus Candida and Aspergillus.
상기 칸디다 속 진균은 칸디다 알비칸스(Candida albicans)이고, 상기 아스퍼질러스 속 진균은 아스퍼질러스 니거(Aspergillus niger)인 것을 특징으로 하는 항균 및 항진균 조성물.The method of claim 3,
The fungus of the genus Candida is Candida albicans, and the fungus of the genus Aspergillus is an antibacterial and antifungal composition, characterized in that the Aspergillus niger (Aspergillus niger).
상기 조성물은 에스체리치아(Escherichia) 속 균주, 슈도모나스(Pseudomonas) 속 균주, 바실러스(Bacillus) 속 균주, 및 스타필로코커스(Staphylococcus) 속 균주에 대해 항균 활성을 갖는 것을 특징으로 하는 항균 및 항진균 조성물.The method of claim 1,
The composition is an antibacterial and antifungal composition, characterized in that it has an antibacterial activity against the strain of the genus Escherichia, Pseudomonas strain, the strain of Bacillus, and the strain of Staphylococcus.
상기 에스체리치아(Escherichia) 속 균주는 에스체리치아 콜라이(Escherichia coli)이고, 상기 슈도모나스(Pseudomonas) 속 균주는 슈도모나스 에루지노사(Pseudomonasaeruginosa)이며, 상기 바실러스(Bacillus) 속 균주는 바실러스 서브틸리스(Bacillus subtilis)이고, 상기 스타필로코커스 속 균주는 스타필로코커스 아우레우스(Staphylococcus aureus)인 것을 특징으로 하는 항균 및 항진균 조성물.The method of claim 5,
The strain of the genus Escherichia is Escherichia coli, the strain of pseudomonas is Pseudomonas aeruginosa, and the strain of the genus Bacillus is Bacillus subtilis Bacillus subtilis), the strain of the genus Staphylococcus is Staphylococcus aureus (Staphylococcus aureus), characterized in that the antibacterial and antifungal composition.
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