KR102034844B1 - Compositions for Anti-Bacterial and Anti-Inflammatory Effect Comprising Callus Extract of Hibiscus Syriacus - Google Patents
Compositions for Anti-Bacterial and Anti-Inflammatory Effect Comprising Callus Extract of Hibiscus Syriacus Download PDFInfo
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- KR102034844B1 KR102034844B1 KR1020190071250A KR20190071250A KR102034844B1 KR 102034844 B1 KR102034844 B1 KR 102034844B1 KR 1020190071250 A KR1020190071250 A KR 1020190071250A KR 20190071250 A KR20190071250 A KR 20190071250A KR 102034844 B1 KR102034844 B1 KR 102034844B1
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- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
- A61K8/9783—Angiosperms [Magnoliophyta]
- A61K8/9789—Magnoliopsida [dicotyledons]
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/105—Plant extracts, their artificial duplicates or their derivatives
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- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
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- A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
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- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q17/00—Barrier preparations; Preparations brought into direct contact with the skin for affording protection against external influences, e.g. sunlight, X-rays or other harmful rays, corrosive materials, bacteria or insect stings
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- A—HUMAN NECESSITIES
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- A61Q19/00—Preparations for care of the skin
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
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Abstract
Description
본 발명은 항균 및 항염증용 조성물에 관한 것으로, 더욱 구체적으로 무궁화 캘러스 추출물을 유효성분으로 포함하는 항균 및 항염증용 조성물에 관한 것이다.The present invention relates to a composition for antibacterial and anti-inflammatory, and more particularly to a composition for antimicrobial and anti-inflammatory comprising an extract of Rose of Sharon as an active ingredient.
생물체의 항상성(homeostasis)을 유지하기 위한 과정에서 중요한 역할을 담당하는 물질들 중 일부는 각종 생물체 유래의 생리 활성 물질이다. 지금까지 수많은 생리활성 물질에 관해 많은 연구가 진행되고 있으며, 그 중, 항균 및 항진균 물질에 관한 연구는 생명과학 및 의학 분야에서 매우 중요한 영역이다.Some of the substances that play an important role in the process of maintaining the homeostasis of organisms are bioactive substances from various organisms. Many studies have been conducted on many bioactive substances, and among them, research on antibacterial and antifungal substances 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 생물체에 관해서 많은 연구가 이루어지고 있으며, 항균 및 항진균 물질을 생산하는 생물체 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 organisms producing antimicrobial and antifungal substances. Research into the development of antibacterial or antifungal agents has long been attempted.
항균물질은 유해 미생물을 죽이면서 인체 또는 동물에게는 독성이 낮고 체내의 효소 등에 의해 비활성화되지 않는 선택적 독성작용(selective toxicity)을 갖는 물질로서, 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 harmful microorganisms and is not toxic to humans or animals and is not inactivated by enzymes in the body. This is mainly due to DNA replication, transcription and decoding of genetic information, and electronic energy. Inhibition of transport, biosynthesis of cell walls, and the like through the mechanism of inhibiting the growth of microorganisms.
현재까지 개발된 주된 항균물질들은 식물, 미생물 등 천연에서 유래한 것과 화학적으로 합성된 것으로 나뉠 수 있는데, 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 substances developed to date can be divided into those derived from nature such as plants and microorganisms. Many antimicrobial substances derived from nature or chemically are used for the treatment of diseases caused by harmful bacteria. Due to the increased resistance to chemically synthesized antimicrobials, there is a growing interest in the development of antimicrobials using antimicrobials derived from nature.
또한, 염증 반응은 세포 또는 조직의 손상이나 박테리아, 곰팡이, 바이러스, 다양한 종류의 알레르기 유발물질 등의 외부감염원에 감염되었을 때 국소 혈관과 체액 중 각종 염증매개인자 및 면역세포가 관련되어, 효소 활성화, 염증 매개물질 분비, 체액 침윤, 세포 이동, 조직 파괴 등 일련의 복합적인 생리적 반응과 홍반, 부종, 발열, 통증 등 외적 증상을 나타낸다. 평상시의 염증 반응은 외부감염원을 제거하고 손상된 조직을 재생하여 생명체 기능회복작용을 하지만, 항원이 제거되지 않거나 내부물질이 원인이 되어 염증 반응이 과도하거나 지속해서 일어나면 오히려 점막 손상을 촉진하고, 그 결과 일부에서는 암 발생 등의 질환을 이끈다. In addition, the inflammatory response is associated with various inflammatory mediators and immune cells in local blood vessels and body fluids when cells or tissues are damaged or when they are infected with external infectious agents such as bacteria, fungi, viruses, and various types of allergens. It exhibits a series of complex physiological reactions including secretion of inflammatory mediators, fluid infiltration, cell migration, and tissue destruction, as well as external symptoms such as erythema, edema, fever, and pain. The usual inflammatory response is to restore the function of life by removing external sources of infection and regenerating damaged tissues.However, if the inflammatory response occurs excessively or continuously due to the absence of antigens or internal substances, it promotes mucosal damage. Some lead to diseases such as cancer.
염증의 발생 원인으로서는 다양한 생화학적인 현상이 관여하고 있는 것으로 알려져 있는데, 특히 일산화질소(Nitric Oxide; NO)를 발생시키는 효소인 산화질소 합성 효소(Nitric Oxide Synthase; NOS)와 프로스타글란딘의 생합성과 관련된 효소들이 염증 반응을 매개하는 데 있어서 가장 중요한 역할을 하는 것으로 알려져 있다. 따라서 L-아르기닌으로부터 일산화질소를 생성시키는 효소인 산화질소 합성효소(NOS)나, 아라키돈산으로부터 프로스타글란딘 등을 합성하는데 관련된 효소인 사이클로옥시게나아제(COX)의 활성 저해를 통해 염증을 차단하는 것이 염증 치료제 개발에 목표가 되고 있다. Inflammation is known to be involved in various biochemical phenomena, especially enzymes related to the biosynthesis of nitric oxide synthase (NOS) and prostaglandins, enzymes that produce nitric oxide (NO) It is known to play the most important role in mediating the inflammatory response. Therefore, blocking inflammation through inhibition of the activity of nitric oxide synthase (NOS), an enzyme that produces nitric oxide from L-arginine, and cyclooxygenase (COX), an enzyme involved in synthesizing prostaglandins from arachidonic acid It is aimed at developing therapeutics.
한편, 우리나라의 국화인 무궁화(Mugunghwa; Hibiscus syriacus L.)는 국내에서만 약 200 여종이 있는 것으로 알려져 있으며, 목근피(Hibiscus syriacus bark)는 무궁화의 뿌리 및 줄기 껍질 부분으로, 예로부터 한방에서 소염, 항균 등의 효능을 가진 약용식물로 이용되고 있다. 그러나, 무궁화가 우리나라의 국화임에도 불구하고 현재까지 국내에서는 무궁화의 재배 및 육종에 관한 연구만 주로 이루어지고 있다. 또한, 무궁화를 이용한 다양한 생리활성 연구가 미흡하며, 특히 생활 속에 적용 가능한 기능성 화장품 또는 식품 등의 원료로 사용 가능한 연구 결과는 거의 없다.On the other hand, Mugunghwa (Higucus syriacus L.), the national flower of Korea, is known to have about 200 species in Korea alone, and the root and stem bark of Mugunghwa is the anti-inflammatory, It is used as a medicinal plant having an antibacterial effect. However, although Mugunghwa is the national flower of Korea, so far, only studies on the cultivation and breeding of Mugunghwa in Korea have been conducted. In addition, research on various physiological activities using hibiscus vulgaris is inadequate, and there are few studies that can be used as raw materials, such as functional cosmetics or food, which can be applied in everyday life.
이러한 배경 하에서, 본 발명자들은 항균 및 염증 개선 효과가 우수한 물질에 관하여 연구를 수행하였으며, 특히 국내 자생 식물체 중 무궁화의 항균 및 염증 개선 효과에 주목하여 이의 항균 및 염증 개선 효능을 더욱 개선하고자 예의 연구노력한 결과, 무궁화 캘러스 추출물이 갖는 우수한 항균 및 항염 효과를 규명하여, 본 발명을 완성하게 되었다.Under these backgrounds, the present inventors have conducted research on substances having excellent antibacterial and inflammation improving effects, and in particular, paying attention to the antimicrobial and inflammation improving effects of Mugunghwa among Korean native plants, the research efforts have been made to further improve its antimicrobial and inflammation improving effects. As a result, the excellent antimicrobial and anti-inflammatory effects of the hibiscus callus extract have been identified and the present invention has been completed.
따라서, 본 발명의 주된 목적은 무궁화 캘러스 추출물을 유효성분으로 포함하는 항균 및 항염증용 화장료 조성물을 제공하는 것이다.Therefore, the main object of the present invention is to provide an antimicrobial and anti-inflammatory cosmetic composition comprising the green leaf callus extract as an active ingredient.
본 발명의 다른 목적은 무궁화 캘러스 추출물을 유효성분으로 포함하는 항균 및 항염증용 약학적 조성물을 제공하는 것이다.Another object of the present invention to provide an antimicrobial and anti-inflammatory pharmaceutical composition comprising the green leaf callus extract as an active ingredient.
본 발명의 또 다른 목적은 무궁화 추출물을 유효성분으로 포함하는 항균 및 항염증용 식품 조성물을 제공하는 것이다.Still another object of the present invention is to provide an antimicrobial and anti-inflammatory food composition comprising the Rose of Sharon extract as an active ingredient.
본 발명의 다른 목적 및 이점은 하기의 발명의 상세한 설명, 청구범위 및 도면에 의해 보다 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 무궁화(Hibiscus Syriacus) 캘러스 추출물을 유효성분으로 포함하는 항균 및 항염증용 화장료 조성물을 제공한다.According to an aspect of the present invention, the present invention provides a cosmetic composition for antibacterial and anti-inflammatory comprising Hibiscus Syriacus callus extract as an active ingredient.
본 발명자들은 유해 세균들과 프로피오니박테리움 아크네스(Propionibacterium acnes) 및 스타필로코커스 에피더미디스(Staphylococcus epidermidis)와 같은 여드름의 원인균을 효과적으로 억제할 수 있는 물질을 찾고자 여러 가지 천연물질들을 대상으로 그 효과를 탐색하였고, 국내 자생 식물체 중 무궁화의 항균 및 항염증 효과에 주목하여 그 효능을 더욱 개선하고자 연구노력한 결과, 무궁화 캘러스 추출물의 항균 및 항염증 활성이 매우 우수함을 발견하였고, 또한 피부에 적용시에도 안전성에 문제가 없음을 확인하였다.The inventors have found that harmful bacteria and propionibacterium acnes In order to find a substance that can effectively suppress acne-causing bacteria such as acnes ) and Staphylococcus epidermidis , we searched for the effects of various natural substances. As a result of efforts to further improve the efficacy by paying attention to the effect, it was found that the antimicrobial and anti-inflammatory activity of the hibiscus callus extract was very excellent, and also confirmed that there is no safety problem even when applied to the skin.
본 발명의 "무궁화(Hibiscus syriacus)"는 한반도 전역에서 자생하는 쌍떡잎식물 아욱목 아욱과의 낙엽관목을 의미한다." Hibiscus syriacus " of the present invention refers to the deciduous shrub of the dicotyledonous plant mallow mallow native to the whole Korean peninsula.
본 발명의 용어 "캘러스(callus)"란, 식물의 전체 또는 일부 조직을 적당한 조건 아래에서 배양하여 분열 증식시킬 수 있는 무정형의 세포괴를 의미한다. 상기 캘러스는 분화기능이 없고 탈분화된 상태이지만, 정기적으로 이식하여 배양하면 그대로 무한 증식할 수 있을 뿐만 아니라, 일정한 조건으로 배양하면 싹, 뿌리, 배 또는 완전한 식물체까지 재분화시킬 수 있다. 이처럼 캘러스를 배양하기 위한 배지에는 옥신이나 시토키닌(cytokinin) 등의 식물호르몬이 필수적으로 포함되어야 하고, 특히 액체배지에서 배양하면 단세포로 분리되기 쉽고 배지나 세포 밀도를 조절하면 단일세포 유래의 세포 집단(clone)을 얻을 수가 있다.As used herein, the term "callus" refers to an amorphous cell mass capable of dividing and proliferating whole or part of a plant under appropriate conditions. The callus has no differentiation and dedifferentiated state, but if transplanted and cultured regularly, it can not only proliferate intact, but also can re-differentiate shoots, roots, pears or even complete plants if cultured under constant conditions. As such, the medium for culturing callus should include plant hormones such as auxin or cytokinin, and in particular, when cultured in a liquid medium, it is easy to separate into single cells. clone).
본 발명에서 사용되는 용어, "추출물"은 식물의 추출 처리에 의하여 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, 액상 또는 건조 분말 형태로 제조되어 사용될 수 있다. As used herein, the term "extract" refers to an extract obtained by extracting 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, a mixture thereof, or the like, It includes the extract itself and extracts of all formulations that can be formed using the extract. The extract of the present invention may preferably be prepared after extraction, prepared in liquid or dry powder form.
본 발명의 상기 무궁화 캘러스의 추출에 있어서, 상기 추출물을 추출하는 방법은 특별히 제한되지 아니하며, 당해 기술 분야에서 통상적으로 사용하는 방법에 따라 추출할 수 있다. 상기 추출 방법의 비제한적인 예로는, 열수 추출법, 초음파 추출법, 여과법, 환류 추출법 등을 들 수 있으며, 이들은 단독으로 수행되거나 2종 이상의 방법을 병용하여 수행될 수 있다. In the extraction of the green light callus of the present invention, the method of extracting the extract is not particularly limited and may be extracted according to a method commonly used in the art. Non-limiting examples of the extraction method, hot water extraction method, ultrasonic extraction method, filtration method, reflux extraction method, and the like, these may be performed alone or in combination of two or more methods.
본 발명에서 상기 무궁화 캘러스를 추출하는데 사용되는 추출 용매의 종류는 특별히 제한되지 아니하며, 당해 기술 분야에서 공지된 임의의 용매를 사용할 수 있다. 상기 추출 용매의 비제한적인 예로는 물; 메탄올, 에탄올, 프로필알코올, 부틸알코올 등의 C1 내지 C4의 저급 알코올; 글리세린, 부틸렌글리콜, 프로필렌글리콜 등의 다가 알코올; 및 메틸아세테이트, 에틸아세테이트, 아세톤, 벤젠, 헥산, 디에틸에테르, 디클로로메탄 등의 탄화수소계 용매; 또는 이들의 혼합물을 사용할 수 있다. In the present invention, the type of the extraction solvent used for extracting the green light callus is not particularly limited, and any solvent known in the art may be used. Non-limiting examples of such extraction solvents include water; C1 to C4 lower alcohols such as methanol, ethanol, propyl alcohol, and butyl alcohol; Polyhydric alcohols such as glycerin, butylene glycol, and propylene glycol; Hydrocarbon solvents such as methyl acetate, ethyl acetate, acetone, benzene, hexane, diethyl ether and dichloromethane; Or mixtures thereof.
본 발명의 항균 및 항염증용 화장료 조성물은 다양한 유해균에 대해 항균 활성이 있다. 본 발명의 항균 및 항염증용 화장료 조성물은 그람 양성균, 그람 음성균에 대하여 항균 활성을 갖고 있는데, 구체적으로 대장균(Escherichia coli), 슈도모나스 에루지노사(Pseudomonas aeruginosa), 스타필로코커스 아우레우스(Staphylococcus aureus), 칸디다 알비칸스(Candida albicans), 아스퍼질러스 나이거(Aspergillus niger) 등의 유해 세균에 대하여 항균 활성을 나타낸다.The antimicrobial and anti-inflammatory cosmetic composition of the present invention has antimicrobial activity against various harmful bacteria. The antimicrobial and anti-inflammatory cosmetic composition of the present invention has antimicrobial activity against Gram-positive bacteria and Gram-negative bacteria, specifically Escherichia coli ( Esherichia coli ), Pseudomonas aeruginosa), Staphylococcus aureus (Staphylococcus aureus), Candida albicans (Candida albicans ), Aspergillus niger ( Aspergillus niger ) and the antimicrobial activity.
본 발명의 항균 및 항염증용 화장료 조성물은 여드름 원인균에 대하여 직접적인 항균 활성이 있는데, 특히, 여드름 유발에 가장 큰 원인으로 지목되고 있는 프로피오니박테리움 속 균주 및 스타필로코커스 속 균주에 대해 항균 활성을 나타내며, 보다 구체적으로는 프로피오니박테리움 아크네스(Propionibacterium acnes) 및 스타필로코커스 에피더미디스(Staphylococcus epidermidis) 균주에 대해 항균 활성을 갖는다. The antimicrobial and anti-inflammatory cosmetic composition of the present invention has a direct antimicrobial activity against acne causing bacteria, in particular, the antimicrobial activity against the strains of the genus Propionibacterium and staphylococcus are considered as the biggest cause of acne. More specifically, it has antimicrobial activity against Propionibacterium acnes and Staphylococcus epidermidis strains.
또한, 본 발명의 항균 및 항염증용 화장료 조성물에서, 상기 항염증 활성은 일산화질소(NO) 생성 억제를 통해 수행되는데, 본 발명자들은 무궁화 캘러스 추출물이 염증 유발물질인 LPS에 의해 매개되는 신호전달체계에서 일산화질소(Nitric Oxide; NO)의 생성 억제를 통해 염증 관련 세포 내 신호전달체계를 효과적으로 차단함으로써 염증성 질환의 예방 또는 치료에 사용될 수 있음을 확인한 바 있다(도 2 참조). In addition, in the antimicrobial and anti-inflammatory cosmetic composition of the present invention, the anti-inflammatory activity is carried out through the inhibition of the production of nitric oxide (NO), the present inventors signal transmission system mediated by the LPS which is an inflammable substance callus extract Inhibiting the production of nitric oxide (Nitric Oxide) in the blocking the inflammation-associated intracellular signaling system has been confirmed that it can be used for the prevention or treatment of inflammatory diseases (see Figure 2).
본 발명의 항균 및 항염증용 화장료 조성물에서, 상기 조성물은 마스크팩, 용액, 외용연고, 크림, 폼, 영양화장수, 유연화장수, 유연수, 유액, 메이크업 베이스, 에센스, 비누, 액체 세정료, 입욕제, 선스크린 크림, 선오일, 현탁액, 유탁액, 페이스트, 겔, 로션, 파우더, 비누, 계면활성제-함유 클린싱, 오일, 분말 파운데이션, 유탁액 파운데이션, 왁스 파운데이션, 패치 및 스프레이로 구성된 군으로부터 선택되는 제형으로 제조할 수 있으나, 이에 제한되는 것은 아니다.In the antimicrobial and anti-inflammatory cosmetic composition of the present invention, the composition is a mask pack, solution, external ointment, cream, foam, nourishing longevity, softening longevity, softening water, emulsion, makeup base, essence, soap, liquid cleansing agent, bathing agent, Formulations selected from the group consisting of sunscreen creams, sun oils, suspensions, emulsions, pastes, gels, lotions, powders, soaps, surfactant-containing cleansing, oils, powder foundations, emulsion foundations, wax foundations, patches and sprays It may be prepared as, but is not limited thereto.
또한, 본 발명의 항균 및 항염증용 화장료 조성물은 일반 피부 화장료에 배합되는 화장품학적으로 허용 가능한 담체를 1종 이상 추가로 포함할 수 있으며, 통상의 성분으로 예를 들면 유분, 물, 계면활성제, 보습제, 저급 알콜, 증점제, 킬레이트제, 색소, 방부제, 향료 등을 적절히 배합할 수 있으나, 이에 제한되는 것은 아니다. In addition, the antimicrobial and anti-inflammatory cosmetic composition of the present invention may further include one or more cosmetically acceptable carriers formulated in general skin cosmetics, and may include, for example, oil, water, surfactants, Moisturizers, lower alcohols, thickeners, chelating agents, pigments, preservatives, fragrances and the like may be appropriately blended, but is not limited thereto.
본 발명의 항균 및 항염증용 화장료 조성물에 포함되는 화장품학적으로 허용 가능한 담체는 제형에 따라 다양하다.The cosmetically acceptable carrier included in the antimicrobial and anti-inflammatory cosmetic composition of the present invention varies depending on the dosage form.
본 발명의 제형이 연고, 페이스트, 크림 또는 젤인 경우에는, 담체 성분으로서 동물성 유, 식물성 유, 왁스, 파라핀, 전분, 트라칸트, 셀룰로오스 유도체, 폴리에틸렌 글리콜, 실리콘, 벤토나이트, 실리카, 탈크, 산화아연 또는 이들의 혼합물이 이용될 수 있다.When the formulation of the present invention is an ointment, paste, cream or gel, the carrier component may be animal oil, vegetable oil, wax, paraffin, starch, tracant, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc, zinc oxide or Mixtures of these may be used.
본 발명의 제형이 파우더 또는 스프레이인 경우에는, 담체 성분으로서 락토스, 탈크, 실리카, 알루미늄 히드록사이드, 칼슘 실케이트, 폴리아미드 파우더 또는 이들의 혼합물이 이용될 수 있고, 특히 스프레이인 경우에는 추가적으로 클로로플루오로히드로카본, 프로판/부탄 또는 디메틸 에테르와 같은 추진제를 포함할 수 있다.If the formulation of the invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate, polyamide powder or mixtures thereof may be used, in particular in the case of a spray additionally chloro Propellants, such as fluorohydrocarbons, propane / butane or dimethyl ether.
본 발명의 제형이 용액 또는 유탁액인 경우에는, 담체 성분으로서 용매, 용해화제 또는 유탁화제가 이용되며, 예컨대 물, 에탄올, 이소프로판올, 에틸 카보네이트, 에틸 아세테이트, 벤질 알콜, 벤질 벤조에이트, 프로필렌글리콜, 1,3-부틸글리콜 오일이 이용될 수 있으며, 특히, 목화씨 오일, 땅콩 오일, 옥수수 배종 오일, 올리브오일, 피마자 오일 및 참깨 오일, 글리세롤 지방족 에스테르, 폴리에틸렌 글리콜 또는 소르비탄의 지방산 에스테르가 이용될 수 있다.When the formulation of the present invention is a solution or emulsion, solvents, solubilizers or emulsions are used as carrier components, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylglycol oil may be used, in particular cottonseed oil, peanut oil, corn seed oil, olive oil, castor oil and sesame oil, fatty acid esters of glycerol aliphatic ester, polyethylene glycol or sorbitan may be used. have.
본 발명의 제형이 현탁액인 경우에는, 담체 성분으로서 물, 에탄올 또는 프로필렌 글리콜과 같은 액상의 희석제, 에톡실화 이소스테아릴 알콜, 폴리옥시에틸렌 소르비톨 에스테르 및 폴리옥시에틸렌 소르비탄 에스테르와 같은 현탁제, 미소결정성 셀룰로오스, 알루미늄 메타히드록시드, 벤토나이트, 아가 또는 트라칸트 등이 이용될 수 있다.When the formulation of the present invention is a suspension, liquid carrier diluents such as water, ethanol or propylene glycol, suspending agents such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, micro Crystalline cellulose, aluminum metahydroxy, bentonite, agar or tracant and the like can be used.
본 발명의 제형이 비누인 경우에는 담체 성분으로서 지방산의 알칼리 금속 염, 지방산 헤미에스테르 염, 지방산 단백질 히드롤리제이트, 이세티오네이트, 라놀린 유도체, 지방족 알콜, 식물성 유, 글리세롤, 당 등이 이용될 수 있다.When the formulation of the present invention is a soap, alkali metal salts of fatty acids, fatty acid hemiester salts, fatty acid protein hydrolyzates, isethionates, lanolin derivatives, aliphatic alcohols, vegetable oils, glycerol, sugars and the like may be used as carrier components. Can be.
본 발명의 항균 및 항염증용 화장료 조성물에서, 무궁화 캘러스 추출물은 구체적으로는 건조 중량으로, 화장료 조성물 총 중량의 0.0001 내지 50 중량%로 포함될 수 있으며, 보다 구체적으로는 0.0005 내지 10 중량%로 포함될 수 있다. 상기 범위 내에서 항균 및 항염증에 대한 우수한 효능을 나타내는 이점이 있으며, 조성물의 제형이 안정화되는 이점이 있다.In the antimicrobial and anti-inflammatory cosmetic composition of the present invention, the green light callus extract may be specifically included as a dry weight, 0.0001 to 50% by weight of the total weight of the cosmetic composition, more specifically may be included as 0.0005 to 10% by weight have. Within this range there is an advantage of showing excellent efficacy against antibacterial and anti-inflammatory, there is an advantage that the formulation of the composition is stabilized.
본 발명의 다른 양태에 따르면, 본 발명은 무궁화(Hibiscus Syriacus) 캘러스 추출물을 유효성분으로 포함하는 항균 및 항염증용 약학적 조성물을 제공한다. 본 발명의 무궁화 캘러스 추출물은 유해 세균에 대한 우수한 항균 활성을 갖는 동시에 염증 발생에 관여하는 일산화질소(Nitric Oxide; NO)의 생성을 효과적으로 억제하여 세균 감염증 및 염증성 질환의 치료 및 예방을 위한 약학적 조성물로 사용될 수 있다. According to another aspect of the present invention, the present invention provides a pharmaceutical composition for antibacterial and anti-inflammatory comprising Hibiscus Syriacus callus extract as an active ingredient. Green light callus extract of the present invention has excellent antibacterial activity against harmful bacteria and at the same time effectively inhibits the production of nitric oxide (Nitric Oxide (NO)) involved in the development of inflammation, pharmaceutical compositions for the treatment and prevention of bacterial infections and inflammatory diseases Can be used as
본 발명에서 "치료"는 상기 조성물의 투여로 세균 감염증 또는 염증성 질환의 증세가 호전되거나 이롭게 되는 모든 행위를 의미하며, "예방"은 상기 조성물의 투여로 세균 감염증 또는 염증성 질환의 발병을 억제 또는 지연시키는 모든 행위를 의미한다.In the present invention, "treatment" means any action that improves or benefits the symptoms of bacterial infection or inflammatory disease by administration of the composition, and "prevention" means to inhibit or delay the onset of bacterial infection or inflammatory disease by administration of the composition. It means all the acts to make.
본 발명에서 "세균 감염증"은 유해 세균에 감염되어 발생하는 질병을 총칭하는 의미로서, 본 발명에서 상기 "세균 감염증"의 비제한적인 예로는 대장균(Escherichia coli), 슈도모나스 에루지노사(Pseudomonas aeruginosa), 스타필로코커스 아우레우스(Staphylococcus aureus), 칸디다 알비칸스(Candida albicans), 아스퍼질러스 나이거(Aspergillus niger) 등의 유해 세균에 감염되어 발생하는 질병을 들 수 있다. 또한, 본 발명의 상기 "세균 감염증"의 비제한적인 예로는 여드름 원인균인 프로피오니박테리움 아크네스(Propionibacterium acnes) 및 스타필로코커스 에피더미디스(Staphylococcus epidermidis) 균주에 감염되어 발생하는 여드름 증상을 들 수 있다. In the present invention, "bacterial infectious disease" is a generic term for a disease caused by infection with harmful bacteria, non-limiting examples of the "bacterial infectious disease" in the present invention E. coli ( Esherichia coli), luge Pseudomonas Labor (Pseudomonas aeruginosa), Staphylococcus aureus (Staphylococcus aureus), Candida albicans (Candida albicans), Aspergillus or disease arising from infection by harmful bacteria, such as this (Aspergillus niger) Can be mentioned. In addition, a non-limiting example of the "bacterial infection" of the present invention is propionibacterium acne causing propionibacterium acnes ) and Staphylococcus epidermidis strains.
또한, 본 발명에서 "염증성 질환"은 염증을 주병변으로 하는 질병을 총칭하는 의미로서, 본 발명에서 상기 염증성 질환은 일산화질소(Nitric Oxide; NO) 매개 염증성 질환인 것을 특징으로 한다. 바람직하게는 상기 일산화질소(Nitric Oxide; NO) 매개 염증성 질환으로는 아토피 피부염, 건선, 부비동염, 비염, 결막염, 천식, 피부염, 염증성 콜라겐 혈관 질환, 사구체신염, 뇌염, 염증성 장염, 만성 폐쇄성 폐질환, 패혈증, 패혈성 쇼크증, 폐섬유증, 미분화 척추관절증, 미분화 관절병증, 관절염, 염증성 골용해, 바이러스 또는 박테리아 감염에 의한 만성 염증질환, 대장염, 궤양성 대장염, 염증성 장질환, 타입 1 당뇨병, 타입 2 당뇨병, 관절염, 류마티스 관절염, 반응성 관절염, 골관절염, 공피증, 골다공증, 아테롬성 동맥경화증, 심근염, 심내막염, 심낭염, 낭성 섬유증, 하시모토 갑상선염, 그레이브스병, 나병, 매독, 라임병(Lyme disease), 보렐리아증(Borreliosis), 신경성-보렐리아증, 결핵, 사르코이드증(Sarcoidosis), 루프스, 동창성 루프스, 결핵성 루프스, 루프스 신염, 전신성 홍반성 루프스, 황반변성, 포도막염, 과민대장 증후군, 크론씨병, 쇼그랜 증후군, 섬유근통, 만성피로 증후군, 만성피로 면역부전 증후군, 근육통성 뇌척수염, 근위축성 측삭경화증, 파키슨병, 알츠하이머병, 다발성경화증, 자폐스펙트럼 장애, 주의력결핍 장애, 주의력 결핍 과잉행동장애 등이 있다. In addition, in the present invention, "inflammatory disease" refers to a disease that is mainly a inflammation of the inflammation, the inflammatory disease in the present invention is characterized in that the nitric oxide (Nitric Oxide; NO) mediated inflammatory disease. Preferably, the nitric oxide (NO) mediated inflammatory disease includes atopic dermatitis, psoriasis, sinusitis, rhinitis, conjunctivitis, asthma, dermatitis, inflammatory collagen vascular disease, glomerulonephritis, encephalitis, inflammatory enteritis, chronic obstructive pulmonary disease, Sepsis, Septic shock, Pulmonary fibrosis, Undifferentiated spondyloarthropathy, Undifferentiated arthrosis, Arthritis, Inflammatory osteolysis, Chronic inflammatory disease caused by viral or bacterial infection, Colitis, Ulcerative colitis, Inflammatory bowel disease,
본 발명의 약학적 조성물은 약제학적으로 허용되는 담체를 포함한다. 본 발명의 약학적 조성물에 포함되는 약제학적으로 허용되는 담체는 제제시에 통상적으로 이용되는 것으로서, 락토스, 덱스트로스, 수크로스, 솔비톨, 만니톨, 전분, 아카시아 고무, 인산 칼슘, 알기네이트, 젤라틴, 규산 칼슘, 미세결정성 셀룰로스, 폴리비닐피롤리돈, 셀룰로스, 물, 시럽, 메틸 셀룰로스, 메틸히드록시벤조에이트, 프로필히드록시벤조에이트, 활석, 스테아르산 마그네슘 및 미네랄 오일 등을 포함하나, 이에 한정되는 것은 아니다. 본 발명의 약학적 조성물은 상기 성분들 이외에 윤활제, 습윤제, 감미제, 향미제, 유화제, 현탁제, 보존제 등을 추가로 포함할 수 있다. 적합한 약학적으로 허용되는 담체 및 제제는 Remington's Pharmaceutical Sciences(19th ed., 1995)에 상세히 기재되어 있다.The pharmaceutical composition of the present invention comprises a pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers included in the pharmaceutical compositions of the present invention are those commonly used in the preparation, such as lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia rubber, calcium phosphate, alginate, gelatin, Calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil, and the like It doesn't happen. In addition to the above components, the pharmaceutical composition of the present invention may further include a lubricant, a humectant, a sweetener, a flavoring agent, an emulsifier, a suspending agent, a preservative, and the like. Suitable pharmaceutically acceptable carriers and formulations are described in detail in Remington's Pharmaceutical Sciences (19th ed., 1995).
본 발명의 약학적 조성물은 경구 또는 비경구 투여할 수 있으며, 비경구 투여인 경우에는 정맥 내 주입, 피하주입, 근육 주입, 복강 주입, 경피 투여 등으로 투여할 수 있다.The pharmaceutical composition of the present invention may be administered orally or parenterally, and in the case of parenteral administration, it may be administered by intravenous injection, subcutaneous injection, intramuscular injection, intraperitoneal injection, transdermal administration, or the like.
본 발명의 약학적 조성물의 적합한 투여량은 제제화 방법, 투여 방식, 환자의 연령, 체중, 성, 병적 상태, 음식, 투여 시간, 투여 경로, 배설 속도 및 반응 감응성과 같은 요인들에 의해 다양하게 처방될 수 있다. 본 발명의 약제학적 조성물의 1일 투여량은 예컨대 0.001-100㎎/㎏이다.Suitable dosages of the pharmaceutical compositions of the present invention may be prescribed in various ways depending on factors such as the formulation method, mode of administration, age, weight, sex, morbidity, food, time of administration, route of administration, rate of excretion, and response to response of the patient. Can be. The daily dose of the pharmaceutical composition of the present invention is, for example, 0.001-100 mg / kg.
본 발명의 약학적 조성물은 당해 발명이 속하는 기술 분야에서 통상의 지식을 가진 자가 쉽게 실시할 수 있는 방법에 따라, 약학적으로 허용되는 담체 및/또는 부형제를 이용하여 제제화함으로써 단위 용량 형태로 제조되거나 또는 다용량 용기 내에 내입시켜 제조될 수 있다. 이때 제형은 오일 또는 수성 매질 중의 용액, 현탁액, 시럽제 또는 유화액 형태이거나 엑스제, 산제, 분말제, 과립제, 정제 또는 캅셀제 형태일 수도 있으며, 분산제 또는 안정화제를 추가로 포함할 수 있다.The pharmaceutical compositions of the present invention may be prepared in unit dosage form by formulating with a pharmaceutically acceptable carrier and / or excipient, according to methods readily available to those skilled in the art. Or by incorporating into a multi-dose container. The formulation may be in the form of solutions, suspensions, syrups or emulsions in oils or aqueous media or in the form of extracts, powders, powders, granules, tablets or capsules, and may further comprise a dispersing or stabilizing agent.
본 발명의 다른 양태에 따르면, 본 발명은 세균 감염증 또는 염증성 질환의 치료 및 예방용 약학적 조성물을 인간을 제외한 개체에 투여하는 단계를 포함하는 세균 감염증 또는 염증성 질환의 예방 또는 치료 방법을 제공한다. 본 발명의 세균 감염증 또는 염증성 질환 및 투여 방법은 상기 본 발명의 약학적 조성물과 관련하여 설명한 바와 같다. 본 발명의 용어 "투여"란 적절한 방법으로 개체에게 소정의 물질을 도입하는 것을 의미한다. 본 발명의 용어 "개체"란 세균 감염증 또는 염증성 질환이 발병하였거나 발병할 수 있는 인간을 포함한 쥐, 생쥐, 가축 등의 모든 동물을 의미한다. 구체적인 예로, 인간을 포함한 포유동물일 수 있다.According to another aspect of the present invention, the present invention provides a method for preventing or treating a bacterial infection or inflammatory disease comprising administering a pharmaceutical composition for treating and preventing a bacterial infection or inflammatory disease to a subject other than a human. The bacterial infection or inflammatory disease of the present invention and the method of administration are as described above in connection with the pharmaceutical composition of the present invention. The term "administration" of the present invention means introducing a certain substance into an individual in a suitable manner. The term "individual" of the present invention means all animals, such as rats, mice, and livestock, including humans, who may or may have developed bacterial infections or inflammatory diseases. As a specific example, it may be a mammal including a human.
또한, 본 발명의 다른 양태에 따르면, 본 발명은 무궁화(Hibiscus Syriacus) 캘러스 추출물을 유효성분으로 포함하는 항균 및 항염증용 식품 조성물을 제공한다. 본 발명의 무궁화 캘러스 추출물은 유해 세균에 대한 우수한 항균 활성을 갖는 동시에 염증 발생에 관여하는 일산화질소(Nitric Oxide; NO)의 생성을 효과적으로 억제하여 세균 감염증 및 염증성 질환의 예방을 위한 식품 조성물로 사용될 수 있다. 본 발명의 무궁화 캘러스 추출물과, 세균 감염증 및 염증성 질환의 치료 및 예방은 상기에서 설명한 바와 같다.In addition, according to another aspect of the present invention, the present invention provides a food composition for antibacterial and anti-inflammatory comprising Hibiscus Syriacus callus extract as an active ingredient. The green light callus extract of the present invention can be used as a food composition for the prevention of bacterial infections and inflammatory diseases by effectively inhibiting the production of nitric oxide (NO) involved in the development of inflammation while having excellent antibacterial activity against harmful bacteria. have. The green light callus extract of the present invention and the treatment and prevention of bacterial infections and inflammatory diseases are as described above.
본 발명의 식품 조성물은 무궁화 캘러스 추출물 이외에 식품 제조 시에 통상적으로 첨가되는 성분을 포함한다. 예를 들어, 단백질, 탄수화물, 지방, 영양소, 조미제 및 감미제를 포함한다. 상술한 탄수화물의 예는 모노사카라이드, 예를 들어, 포도당, 과당 등; 디사카라이드, 예를 들어 말토스, 슈크로스, 올리고당 등; 및 폴리사카라이드, 예를 들어 덱스트린, 사이클로덱스트린 등과 같은 통상적인 당 및 자일리톨, 소르비톨, 에리트리톨 등의 당알코올이다. 감미제로서 천연 감미제[타우마틴, 스테비아 추출물(예를 들어 레바우디오시드 A, 글리시리진 등)] 및 합성 감미제(사카린, 아스파르탐 등)를 사용할 수 있다. 예컨대, 본 발명의 식품 조성물이 드링크제로 제조되는 경우에는 본 발명의 유효성분 이외에 구연산, 액상과당, 설탕, 포도당, 초산, 사과산, 과즙, 두충 추출액, 대추 추출액, 감초 추출액 등을 추가로 포함할 수 있다.The food composition of the present invention contains components that are commonly added during food preparation in addition to the green light callus extract. Examples include proteins, carbohydrates, fats, nutrients, seasonings and sweeteners. Examples of the above carbohydrates include monosaccharides such as glucose, fructose and the like; Disaccharides such as maltose, sucrose, oligosaccharides and the like; And sugars such as conventional sugars such as polysaccharides such as dextrin, cyclodextrin and the like and xylitol, sorbitol, erythritol. As sweeteners, natural sweeteners (tauumatin, stevia extract (eg rebaudioside A, glycyrrhizin, etc.)) and synthetic sweeteners (saccharin, aspartame, etc.) can be used. For example, when the food composition of the present invention is prepared with a drink, in addition to the active ingredient of the present invention, citric acid, liquid fructose, sugar, glucose, acetic acid, malic acid, fruit juice, tofu extract, jujube extract, licorice extract, etc. may be further included. have.
본 발명의 특징 및 이점을 요약하면 다음과 같다:The features and advantages of the present invention are summarized as follows:
(a) 본 발명의 무궁화 캘러스 추출물을 포함하는 조성물은 다양한 유해세균 및 여드름 원인균들에 대해 폭넓은 항균 스펙트럼을 나타내고 있으며, 본 발명의 조성물은 유해세균에 의해 발생하는 다양한 세균 감염증에 적용이 가능하다. (a) The composition containing the green light callus extract of the present invention exhibits a broad antimicrobial spectrum against various harmful bacteria and acne causing bacteria, the composition of the present invention can be applied to various bacterial infections caused by harmful bacteria .
(b) 또한, 본 발명의 무궁화 캘러스 추출물을 포함하는 조성물은 세포 독성이 없고, 산화질소(NO) 매개 염증 반응을 효과적으로 억제하여 염증성 질환의 치료 및 예방에 효과적으로 사용될 수 있다. (b) In addition, the composition comprising the green light callus extract of the present invention is not cytotoxic and effectively inhibits nitric oxide (NO) mediated inflammatory response can be effectively used for the treatment and prevention of inflammatory diseases.
(c) 따라서, 본 발명의 조성물은 화장품, 의약, 건강기능식품 등의 다양한 분야에서 항균 및 항염증 목적으로 유용하게 이용가능하다.(c) Therefore, the composition of the present invention can be usefully used for antibacterial and anti-inflammatory purposes in various fields, such as cosmetics, medicine, health food.
도 1은 무궁화 캘러스 추출물을 인간각질형성세포에 처리한 후 세포 생존율의 변화를 측정하여 나타낸 그래프이다.
도 2는 무궁화 캘러스 추출물을 RAW 264.7 세포에 처리한 후 NO 생성량의 변화를 측정하여 나타낸 그래프이다.Figure 1 is a graph showing the measurement of the change in cell viability after treatment with the green keratin extract human rose keratinocytes.
Figure 2 is a graph showing the measurement of the change in the amount of NO production after processing the green leaf callus extract to RAW 264.7 cells.
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.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.
실시예Example 1. 무궁화 1. Mugunghwa 캘러스Callus 추출물의 제조 Preparation of Extract
무궁화 잎을 기내(in vitro)에서 무균 상태로 배양하기 위해 70% 에탄올과 4% 차아염소산나트륨을 사용하여 1분간 소독한 후, 멸균수로 여러 번 세척하였다. 핀셋과 메스를 사용하여 무균 상태의 무궁화 잎을 잘라 상처를 낸 후, 1ppm 2,4-D(2,4 Dichlorophenoxyacetic Acid), 및 0.5ppm BA(6-benzylaminopurine) 호르몬이 포함된 3% Sucrose WPM(woody plant medium) 고체배지에 치상하여 4주간 배양한 후, 주기적으로 계대배양하였다.Green leaves were sterilized with 70% ethanol and 4% sodium hypochlorite for 1 minute to sterile in vitro, and then washed several times with sterile water. Using tweezers and a scalpel, cut the sterile green leaves and cut them, and then cut 3% Sucrose WPM with 1 ppm 2,4-D (2,4 Dichlorophenoxyacetic Acid) and 0.5 ppm BA (6-benzylaminopurine) woody plant medium) and then incubated for 4 weeks on a solid medium and then subcultured periodically.
식물생장 호르몬인 2,4-D(2,4 Dichlorophenoxyacetic Acid) 1ppm과 BA(6-benzylaminopurine) 0.5ppm이 첨가된 3% Sucrose WPM 액체배지를 조제하였고, 상기 고체배지에서 계대배양한 캘러스를 상기 액체배지에 접종하였고, 3주간 25℃ 암(dark) 조건에서 배양한 후, 원심분리하여 캘러스를 회수하였다. A 3% Sucrose WPM liquid medium containing 1 ppm of plant growth hormone 2,4-D (2,4 Dichlorophenoxyacetic Acid) and 0.5 ppm of BA (6-benzylaminopurine) was prepared, and the callus cultured in the solid medium was used as the callus. The medium was inoculated, cultured in a dark condition at 25 ° C. for 3 weeks, and then the callus was recovered by centrifugation.
상기 회수된 캘러스에서 수분을 제거한 후 동결건조하였고, 막자사발을 사용하여 파쇄하였다. 파우더 상태의 캘러스를 70% 에탄올(24h, 37℃)로 추출하고, 필터 페이퍼를 사용하여 여과한 후 감압 농축하여, 무궁화 캘러스 추출물을 수득하였다.Water was removed from the recovered callus and lyophilized, and crushed using a mortar and pestle. The powdered callus was extracted with 70% ethanol (24 h, 37 ° C.), filtered using a filter paper, and then concentrated under reduced pressure to obtain a hibiscus callus extract.
실시예Example 2. 무궁화 2. Rose of Sharon 캘러스Callus 추출물의한Extract of 세포 독성 효과 검증 Cytotoxic effect verification
상기 실시예 1에서 수득한 무궁화 캘러스 추출물이 인간각질형성세포(HaCaT, Human immortalized keratinocyte)의 생장에 미치는 효과를 아래와 같은 방법으로 확인하였다. The effect of the hibiscus callus extract obtained in Example 1 on the growth of human immortalized keratinocytes (HaCaT) was confirmed by the following method.
우선, 10% FBS(fetal bovine serum, Cambrex)가 포함된 전용 배지인 DMEM(Dulbeccos modified Eagles medium) 배지에 인간각질형성세포를 24 well 세포 배양 접시에 1×105 농도로 접종한 후 24시간 동안, 37℃, 5% CO2 습윤 조건에서 배양하였다. 이후, 배지를 제거하고 무혈청 DMEM 배지로 희석한 실시예 1에서 제조한 무궁화 캘러스 추출물을 농도별(1, 및 10㎍/mL)로 처리하여 24시간 동안 배양하였다. 24시간 후, 1mg/mL의 MTT를 처리하고 2시간 후, 세포에 MTT 처리로 인해 생성된 포마잔(formazan)을 DMSO로 녹여 570nm에서 흡광도를 측정하였다. 양성대조군으로 TGF-β10㎍/mL를 처리하였다.First, human keratinocytes were inoculated at 1 × 10 5 concentration in a 24 well cell culture dish in DMEM (Dulbeccos modified Eagles medium), a medium containing 10% FBS (fetal bovine serum, Cambrex), for 24 hours. Incubated at 37 ° C., 5% CO 2 wet conditions. Subsequently, the cultured callus extract prepared in Example 1, in which the medium was removed and diluted with serum-free DMEM medium, was treated with concentrations (1, and 10 µg / mL) and incubated for 24 hours. After 24 hours, 1 mg / mL MTT treatment and 2 hours later, formazan (formazan) produced by MTT treatment was dissolved in DMSO, and the absorbance was measured at 570 nm. TGF-
그 결과, 무궁화 캘러스 추출물이 인간각질형성세포의 세포 생존율에 크게 영향을 미치지 않는 것을 확인하였으며, 오히려 1㎍/mL 농도에서 인간각질형성세포의 생존을 촉진하는 결과를 나타냈다. 상기 결과를 통해 무궁화 캘러스 추출물은 인체에 독성을 나타내지 않음을 확인할 수 있었다(도 1 참조).As a result, it was confirmed that the green leaf callus extract does not significantly affect the cell viability of human keratinocytes, but rather promotes the survival of human keratinocytes at a concentration of 1 μg / mL. Through the above results, it was confirmed that the Rose of Sharon callus extract is not toxic to the human body (see FIG. 1).
실시예Example 3. 무궁화 3. Rose of Sharon 캘러스Callus 추출물의 항균 테스트 Antimicrobial Test of Extracts
3-1. 유해 세균에 대한 항균 테스트3-1. Antibacterial test for harmful bacteria
실시예 1에서 제조한 무궁화 캘러스 추출물이 갖는 항균 활성 탐색은 유해 미생물을 이용하여 미생물 생장 최소억제농도(MIC)를 평가하였다.The antimicrobial activity search of the green light callus extract prepared in Example 1 evaluated the microbial growth minimum inhibitory concentration (MIC) using harmful microorganisms.
유해 세균은 대장균(Escherichia coli KCTC 2571), 슈도모나스 에루지노사(Pseudomonas aeruginosa KCTC 2513), 스타필로코커스 아우레우스(Staphylococcus aureus KCTC 3881) 균주, 진균류인 칸디다 알비칸스(Candida albicans KCTC 7965), 및 아스퍼질러스 나이거(Aspergillus niger KCTC 6317)를 사용하였다(KCTC 한국생명공학연구원 생물자원센터).Harmful bacteria include Escherichia coli KCTC 2571, Pseudomonas aeruginosa KCTC 2513, Staphylococcus aureus KCTC 3881 strain, the fungus Candida albicans, and AsC7 7965. Aspergillus niger KCTC 6317 was used (KCTC Bioresource Center, Korea Research Institute of Bioscience and Biotechnology).
[표 1]TABLE 1
대장균(E. coli)의 배양에 사용된 배지는 LB 배지를 사용하였고, 슈도모나스 에루기노사(P. aeruginosa)와 스타필로코커스 아우레우스(S. aureus) 균주의 배양은 NA 배지, 그리고 칸디다 알비칸스(C. albicans)와 아스퍼질러스 나이거(Aspergillus niger) 균주는 각각 YM 배지 및 PDA 배지를 사용하였다. 각 세균은 2~3개의 집락(1×107 CFU/mL)을 얻어 접종하였다. 비교예 1은 1% 메틸파라벤을 사용하였고, 비교예 2는 타사제품 A를 1% 농도(w/w)로 사용하였다.The medium used for the culture of E. coli was LB medium, and P. aeruginosa and S. aureus strains were cultured with NA medium, and Candida albi. C. albicans and Aspergillus niger strains used YM medium and PDA medium, respectively. Each bacterium was inoculated with two to three colonies (1 × 10 7 CFU / mL). Comparative Example 1 used 1% methyl paraben, and Comparative Example 2 used a third-party product A at a 1% concentration (w / w).
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 based on 1 × 10 5 ~ 6 CFU / mL, diluted in the medium and inoculated and maintained for 2 to 3 hours at 37 ℃.
본 실험에 사용한 무궁화 캘러스 추출물은 2% 농도(w/w)로 적정하여 각각 2배 계열로 희석하였다. 각 유해 세균들과 시료가 함유된 배지를 96 well plate에 옮기고 37℃에서 16~24시간 동안 배양한 후, 600nm에서 Multi label Counter(1420, Perkin Elmer)로 측정하여 각 유해 세균들에 대한 무궁화 캘러스 추출물의 최소저해농도 값을 구하고, 이를 하기 [표 2]에 나타내었다.The green rose extract used in this experiment was titrated at 2% concentration (w / w) and diluted in 2-fold series. Transfer medium containing each harmful bacteria and sample to 96 well plate, incubate at 37 ℃ for 16 ~ 24 hours, measure with Multi label Counter (1420, Perkin Elmer) at 600nm and callus callus for each harmful bacteria The minimum inhibitory concentration value of the extract was obtained and shown in Table 2 below.
[표 2]TABLE 2
그 결과, 본 발명의 무궁화 캘러스 추출물은 화학적으로 합성된 항균제인 메틸파라벤과 비교하여 유사한 항균력을 나타내었고, 시중에서 판매되는 타사의 천연항균 제품과 비교할 때 월등하게 우수한 항균 효과를 나타내는 것으로 확인되었다. As a result, the hibiscus callus extract of the present invention showed a similar antimicrobial activity compared to the chemically synthesized antimicrobial agent methylparaben, it was confirmed to exhibit an excellent antimicrobial effect compared to other natural antibacterial products of the market.
3-2. 여드름 유발 원인균에 대한 항균 테스트3-2. Antibacterial test for acne causing bacteria
본 발명의 무궁화 캘러스 추출물이 여드름 원인균에 대한 직접적인 항균 활성이 있는지를 확인하기 위하여 여드름 원인균으로 알려진 프로피오니박테리움 아크네스(Propionibacterium acnes) 균주와 스타필로코커스 에피더미디스(Staphylococcus epidermidis) 균주를 대상으로 여드름 유발 원인균의 생장 억제 효능을 검증하였다. Propionibacterium known as acne causative bacteria in order to determine whether the green light extract of the present invention has direct antimicrobial activity against acne causative bacteria acnes ) strain and Staphylococcus epidermidis strain were tested for growth inhibition of acne-causing bacteria.
[표 3]TABLE 3
검증을 위해 액체배지 희석법에 따라 프로피오니박테리움 아크네스(P. acnes) 균주와 스타필로코커스 에피더미디스 균주에 대한 최소 성장억제농도(Minimun Inhibitory Concentration, MIC)를 구하여 검증하였다.Minimal Inhibitory Concentration (MIC) was determined for the Propionibacterium acnes strain and Staphylococcus epidermidis strain according to the liquid medium dilution method.
먼저, 프로피오니박테리움 아크네스 균주는 RCM 배지에, 스타필로코커스 에피더미디스 균주는 NB 배지를 이용하여 배양하였고, 상기 [표 3]의 배양 조건으로 배양한 후 시험에 사용하였다. 각 균주별 액체 배지와 무궁화 캘러스 추출물을 혼합하여 2종의 균주를 1×106 CFU/ml의 초기 균수로 준비하여 미생물 저감 효능 시험을 실시하였다. 준비된 2종 균주를 대조군, 비교군을 포함하여 각각 접종하여 균주별 최적 조건에서 배양하였으며, 0, 1, 24시간마다 시료를 채취하여 멸균된 식염수를 이용하여 10배수로 희석하여 한천배지에 도말하였다. 도말한 플레이트들은 최적조건에서 배양한 후, 전형적인 집락을 확인 및 계수하여 감소율을 확인하였고, 이를 통해 2종 미생물에 대한 저감 효능을 확인할 수 있었다. 비교 검증을 위해 종래 여드름 원인균에 대해 항균효과가 알려진 0.1%(w/v) Salicylic acid(비교예 3)와 1%(w/w) Tea Tree oil(비교예 4)을 사용하였다.First, the Propionibacterium acnes strain was cultured in RCM medium, and Staphylococcus epidermidis strain was cultured using NB medium, and then used for the test after culturing under the culture conditions of [Table 3]. The microbial reduction efficacy test was performed by mixing two strains with an initial bacterial count of 1 × 10 6 CFU / ml by mixing the liquid medium of each strain and the hibiscus callus extract. The prepared two strains were inoculated, including the control group and the comparison group, and incubated under optimum conditions for each strain. Samples were collected every 0, 1, and 24 hours, and diluted to 10-fold using sterilized saline and plated on agar medium. After culturing the plate at the optimum conditions, the typical colonies were identified and counted to confirm the reduction rate, through which the reduction effect on the two microorganisms could be confirmed. For comparative verification, 0.1% (w / v) Salicylic acid (Comparative Example 3) and 1% (w / w) Tea Tree oil (Comparative Example 4), which are known to have antimicrobial effects against conventional acne-causing bacteria, were used.
[표 4]TABLE 4
그 결과, 상기 [표 4]에 나타난 바와 같이, 본 발명의 무궁화 캘러스 추출물을 시료로 처리한 시험군에서는 여드름 원인균인 프로피오니박테리움 아크네스 균주와 스타필로코커스 에피더미디스 균주의 생장을 99.9% 이상 억제하는 것으로 관찰되었으며, [비교예 3]의 Salicylic acid 또는 [비교예 4]의 Tea Tree oil과 대비하여 여드름 원인균에 대한 생장 억제 효과가 더욱 우수한 것으로 나타났다. 상기 결과를 통해 본 발명의 무궁화 캘러스 추출물이 여드름 피부의 개선에 효과적으로 사용할 수 있음을 확인할 수 있었다.As a result, as shown in [Table 4], in the test group treated with the green light callus extract of the present invention as a sample, 99.9% of the growth of acne-causing bacteria, Propionibacterium acnes strain and Staphylococcus epidermidis strain, was observed. Abnormal inhibition was observed, and growth inhibition effect against acne-causing bacteria was better than that of Salicylic acid of [Comparative Example 3] or Tea Tree oil of [Comparative Example 4]. Through the above results, it was confirmed that the Rose of Sharon callus extract of the present invention can be effectively used to improve acne skin.
실시예Example 4. 무궁화 4. Rose of Sharon 캘러스Callus 추출물의 항염 테스트 Anti-inflammatory Test of Extracts
무궁화 캘러스 추출물의 항염증 효과를 확인하기 위해, 염증 유발에 관여하는 대표적인 세포 독성물질인 일산화질소(nitric oxide, NO)의 생성 억제율을 측정하였다.In order to confirm the anti-inflammatory effect of the green leaf callus extract, the inhibition rate of nitric oxide (NO), which is a representative cytotoxic substance involved in inflammation, was measured.
먼저, 대식세포주인 RAW 264.7 세포를 한국세포주은행(KTCC, Seoul, Korea)에서 분양받아 사용하였다. 구체적으로, 10% FBS(Fetal bovine serum)를 함유한 DMEM(Dulbecco’s modified Eagle’s medium)에 10% FBS, 100㎍/㎖ 페니실린 및 100㎍/㎖ 스트렙토마이신을 첨가한 배지에 상기 세포를 접종하여 37℃, 5% CO2 조건에서 배양하였다.First, RAW 264.7 cells, which are macrophages, were distributed and used by Korea Cell Line Bank (KTCC, Seoul, Korea). Specifically, the cells were inoculated in a medium containing 10% FBS, 100 µg / ml penicillin, and 100 µg / ml streptomycin in Dulbecco's modified Eagle's medium (DMEM) containing 10% FBS (Fetal bovine serum) to 37 ° C. Incubated at 5% CO 2 .
구체적으로, 상기 RAW 264.7 세포를 1×106 cells/mL의 농도(in DMEM)로 96 웰 플레이트에 접종하고 24시간 배양 후, 실시예 1에서 제조된 무궁화 캘러스 추출물을 0.1, 1, 및 10㎍/㎖ 농도로 희석한 시료와 내독소로 알려진 LPS(1μg/ml)를 함유한 새로운 배지를 동시에 처리하여 24시간 배양하였다. 그 후, 세포 배양 상등액 100㎕와 Griess reagent[1%(w/v) sulfanilamide, 0.1%(w/v) naphtylethylenediamine in 2.5%(v/v) phosphoric acid] 100㎕를 혼합하여 96 웰 플레이트에서 10분 동안 반응시키고, ELISA 리더를 사용하여 540nm에서 흡광도를 측정하여 생성된 산화질소의 양을 측정하였다. 생성된 아질산염(nitrite)의 농도는 아질산나트륨(sodium nitrite)을 DMEM 배지에 용해한 표준 곡선을 이용하여 계산하였다. LPS를 처리한 대조군과 LPS를 처리하지 않은 대조군에서 생성된 아질산염의 생성량 차이를 기준으로 각 시료의 일산화질소 생성 저해 활성을 확인하였다.Specifically, the RAW 264.7 cells were inoculated in a 96 well plate at a concentration of 1 × 10 6 cells / mL (in DMEM) and cultured for 24 hours, and then 0.1, 1, and 10 μg of the green light extract prepared in Example 1 were used. Samples diluted at a concentration of / ml and fresh medium containing LPS (1 μg / ml), known as endotoxin, were simultaneously treated and incubated for 24 hours. Subsequently, 100 μl of the cell culture supernatant and 100 μl of Griess reagent [1% (w / v) sulfanilamide, 0.1% (w / v) naphtylethylenediamine in 2.5% (v / v) phosphoric acid] were mixed. The reaction was carried out for minutes and the absorbance was measured at 540 nm using an ELISA reader to determine the amount of nitric oxide produced. The concentration of the resulting nitrite was calculated using a standard curve of sodium nitrite dissolved in DMEM medium. Nitrogen monoxide production inhibitory activity of each sample was confirmed based on the difference in the amount of nitrite produced in the LPS-treated control group and the LPS-treated control group.
실험 결과, 본 발명의 무궁화 캘러스 추출물은 LPS처리 대조군에 비해 각각 25%(0.1㎍/㎖), 33%(1㎍/㎖), 및 63%(10㎍/㎖) 수준으로 산화질소 생성을 억제하여 산화질소 생성 억제에 우수한 효과를 보이는 것으로 관찰되었다(도 2 참조). As a result, the green leaf callus extract of the present invention inhibited nitric oxide production at 25% (0.1 μg / ml), 33% (1 μg / ml), and 63% (10 μg / ml) levels, respectively, compared to the LPS-treated control group. It was observed to show an excellent effect on the inhibition of nitric oxide production (see Fig. 2).
상기와 같은 결과를 통해, 본 발명의 무궁화 캘러스 추출물은 염증반응에 중요한 역할을 하는 산화질소의 생성을 직접적으로 저해하여 염증관련 질환의 치료 및 예방에 효과적으로 작용할 수 있음을 확인할 수 있었다. Through the above results, it was confirmed that the green light callus extract of the present invention can directly inhibit the production of nitric oxide that plays an important role in the inflammatory response can effectively act in the treatment and prevention of inflammation-related diseases.
상기와 같은 결과를 통해, 본 발명의 무궁화 캘러스 추출물을 포함하는 조성물은 세포 독성이 없고, 유해 세균에 대한 우수한 항균 효과가 있으며, 여드름 원인균의 생장을 효과적으로 억제하고, NO 생성을 직접적으로 억제하여 염증성 질환의 치료 및 예방에 효과가 있음을 확인할 수 있었다. Through the above results, the composition containing the green light callus extract of the present invention is not cytotoxic, has excellent antimicrobial effect against harmful bacteria, effectively inhibit the growth of acne causative bacteria, and directly inhibit the production of NO inflammatory It was confirmed that it is effective in the treatment and prevention of the disease.
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.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.
따라서, 본 발명의 실질적인 범위는 첨부된 청구항과 그의 등가물에 의하여 정의된다고 할 것이다.Thus, the substantial scope of the present invention will be defined by the appended claims and equivalents thereof.
Claims (6)
여드름 원인균인 프로피오니박테리움 아크네스(Propionibacterium acnes) 및 스타필로코커스 에피더미디스(Staphylococcus epidermidis) 균주에 항균 활성을 갖는 것을 특징으로 하는,
항균 및 항염증용 화장료 조성물.It contains Hibiscus Syriacus callus extract as an active ingredient,
Characterized by having antibacterial activity against the acne causing bacteria Propionibacterium acnes and Staphylococcus epidermidis strains,
Cosmetic composition for antibacterial and anti-inflammatory.
상기 조성물은 대장균(Escherichia coli), 슈도모나스 에루지노사(Pseudomonas aeruginosa), 스타필로코커스 아우레우스(Staphylococcus aureus), 칸디다 알비칸스(Candida albicans), 및 아스퍼질러스 나이거(Aspergillus niger)로 이루어진 군에서 선택된 하나 이상의 유해 세균에 추가적으로 항균 활성을 갖는 것을 특징으로 하는 항균 및 항염증용 화장료 조성물.The method of claim 1,
The composition is composed of Escherichia coli , Pseudomonas aeruginosa , Staphylococcus aureus , Candida albicans , and Aspergillus niger Antimicrobial and anti-inflammatory cosmetic composition, characterized in that it has an antimicrobial activity in addition to one or more harmful bacteria selected from.
상기 항염증 활성은 일산화질소(NO) 생성 억제를 통해 수행되는 것을 특징으로 하는 항균 및 항염증용 화장료 조성물.The method of claim 1,
The anti-inflammatory activity is characterized in that the antimicrobial and anti-inflammatory cosmetic composition is carried out through the inhibition of nitric oxide (NO) production.
여드름 원인균인 프로피오니박테리움 아크네스(Propionibacterium acnes) 및 스타필로코커스 에피더미디스(Staphylococcus epidermidis) 균주에 항균 활성을 갖는 것을 특징으로 하는,
항균 및 항염증용 약학적 조성물.It contains Hibiscus Syriacus callus extract as an active ingredient,
Characterized by having antibacterial activity against the acne causing bacteria Propionibacterium acnes and Staphylococcus epidermidis strains,
Pharmaceutical composition for antibacterial and anti-inflammatory.
여드름 원인균인 프로피오니박테리움 아크네스(Propionibacterium acnes) 및 스타필로코커스 에피더미디스(Staphylococcus epidermidis) 균주에 항균 활성을 갖는 것을 특징으로 하는,
항균 및 항염증용 식품 조성물.It contains Hibiscus Syriacus callus extract as an active ingredient,
Characterized by having antibacterial activity against the acne causing bacteria Propionibacterium acnes and Staphylococcus epidermidis strains,
Antimicrobial and anti-inflammatory food composition.
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| KR102266125B1 (en) * | 2020-04-23 | 2021-06-17 | (주)에스디생명공학 | Composition for preventing or treating inflammatory diseases and cancer, including Hibiscus syriacus leaf callus extract as an active ingredient |
| KR20220017247A (en) * | 2020-08-04 | 2022-02-11 | (주)에스디생명공학 | Anti-inflammatory composition comprising gold nanoparticles containing Hibiscus syriacus callus extract |
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| Hindawi BioMed Research International., Vol. 2017, pp. 1-9 (2017)* * |
| 인터넷 화장품 판매사이트 Na beauty™ "INGREDIENTS : HIBISCUS SYRIACUS CALLUS EXTRACT" (2017) ※ 주소 : https://nabeauty.com/essential_grid/ingredients-hibiscus-syriacus-callus-extract/* * |
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| KR102266125B1 (en) * | 2020-04-23 | 2021-06-17 | (주)에스디생명공학 | Composition for preventing or treating inflammatory diseases and cancer, including Hibiscus syriacus leaf callus extract as an active ingredient |
| KR20220017247A (en) * | 2020-08-04 | 2022-02-11 | (주)에스디생명공학 | Anti-inflammatory composition comprising gold nanoparticles containing Hibiscus syriacus callus extract |
| KR102397659B1 (en) * | 2020-08-04 | 2022-05-13 | (주)에스디생명공학 | Anti-inflammatory composition comprising gold nanoparticles containing Hibiscus syriacus callus extract |
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