KR102289506B1 - Anti-bacterial composition for Streptococcus mutans and composition for removing halitosis using the same - Google Patents
Anti-bacterial composition for Streptococcus mutans and composition for removing halitosis using the same Download PDFInfo
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- KR102289506B1 KR102289506B1 KR1020190090418A KR20190090418A KR102289506B1 KR 102289506 B1 KR102289506 B1 KR 102289506B1 KR 1020190090418 A KR1020190090418 A KR 1020190090418A KR 20190090418 A KR20190090418 A KR 20190090418A KR 102289506 B1 KR102289506 B1 KR 102289506B1
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
- A61K8/9755—Gymnosperms [Coniferophyta]
- A61K8/9761—Cupressaceae [Cypress family], e.g. juniper or cypress
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
- A61K8/9783—Angiosperms [Magnoliophyta]
- A61K8/9789—Magnoliopsida [dicotyledons]
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- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q11/00—Preparations for care of the teeth, of the oral cavity or of dentures; Dentifrices, e.g. toothpastes; Mouth rinses
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- 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
- A61Q17/005—Antimicrobial preparations
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2300/00—Mixtures or combinations of active ingredients, wherein at least one active ingredient is fully defined in groups A61K31/00 - A61K41/00
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Abstract
본 발명은 측백나무 잎 추출물, 비파나무 잎 추출물 등을 이용한 치아우식증의 원인균인 스트렙토코커스 뮤탄스에 대한 항균 조성물과 이를 이용한 구취 제거용 조성물을 개시한다.The present invention discloses an antibacterial composition for Streptococcus mutans, a causative bacterium of dental caries, and a composition for removing bad breath using the same, using an arborvitae leaf extract, a loquat leaf extract, and the like.
Description
본 발명은 스트렙토코커스 뮤탄스 균에 대한 항균 조성물 및 이를 이용한 구취 제거용 조성물에 관한 것이다. The present invention relates to an antibacterial composition for Streptococcus mutans and a composition for removing bad breath using the same.
국민의 생활수준 향상과 의료기술 발달로 평균수명이 연장되면서 고령인구의 삶의 질을 높이기 위한 구강건강 관리의 중요성이 커지고 있다. 구강건강이 문제가 되어 치아상실까지 유발할 수 있는 대표적인 질환으로 치아우식증과 치주질환이 있다. 2013년 보건복지부의 자료에 따르면, 우리나라 성인 남성의 영구치우식 경험율은 87% 수준이었으며, 여성은 90% 이었다. 또한 우리나라 성인의 만 30세 이상 치주질환 유병률은 2007년부터 2012년까지 감소하다가 2013년에는 다시 증가하였고, 성인 세 명 중에 한명은 치주질환에 이완된 것으로 조사되었다.The importance of oral health management to improve the quality of life of the elderly is increasing as the average life expectancy is extended due to the improvement of people's living standards and the development of medical technology. Dental caries and periodontal disease are typical diseases that can cause tooth loss due to oral health problems. According to data from the Ministry of Health and Welfare in 2013, the experience rate of permanent dental caries among Korean adult males was 87%, and that of females was 90%. In addition, the prevalence of periodontal disease among Korean adults aged 30 years and older decreased from 2007 to 2012, and then increased again in 2013.
치아우식증은 세계인구의 80% 이상이 경험하고 있으며 서구 일부 선진국에서 1970년대 이후 지속적으로 발병이 감소하고 있으나 우리나라를 비롯하여 동유럽과 일부 아프리카 지역 등에서는 아직도 증가하고 있다(Nishi et al., 2002 Community Dent Oral Epidemiol 2002 30:296-301.). 치아우식증은 여러 가지 동통을 유발할 뿐 아니라, 계속적으로 진행되면 치근단, 치조골 파괴 및 치아 상실을 유발한다. 치아우식증 발생은 식이습관 및 구강 내 미생물과 깊은 관련이 있으며 인체 구강에 서식하는 약 750여종에 달하는 미생물 중 치아우식증을 대표하는 원인균은 스트렙토코커스 뮤탄스(Streptococcus mutans)로 알려져 있다(Loesche WJ. Medical Microbiology 1996 (4th ed.);Kilian Clarke J. Br J Exp Pathol. 1924 Jun; 5(3): 141147.; 김혜영, 서울대학교 대학원 박사학위논문 2002). Dental caries is experienced by more than 80% of the world's population, and although the incidence has continuously decreased since the 1970s in some developed countries in the West, it is still increasing in Eastern Europe and some African regions, including Korea (Nishi et al., 2002 Community Dent) (Nishi et al., 2002 Community Dent). Oral Epidemiol 2002 30:296-301.). Dental caries not only causes various pains, but also causes apical, alveolar bone destruction and tooth loss if it continues to progress. The occurrence of dental caries is closely related to dietary habits and oral microorganisms, and the causative bacterium representing dental caries among about 750 types of microorganisms that inhabit the human oral cavity is known as Streptococcus mutans (Loesche WJ. Medical). Microbiology 1996 (4th ed.);Kilian Clarke J. Br J Exp Pathol. 1924 Jun; 5(3): 141147.; Hyeyoung Kim, Seoul National University doctoral thesis 2002).
구강 내에서 스트렙토코커스 뮤탄스가 당분 등을 접하게 되면 치아 사이에 polysaccharide 결정체를 만들어 저장하게 된다. 치아 피막에 부착하여 glucosyltransferase(GTase) 효소의 작용으로 불용성 글루칸(glucan)을 합성한다. 합성된 글루칸은 균 사이의 결합력을 증가시키고 신진대사 폐기물로 형성된 젖산이 polysaccharide 결정체 아래에 축적하여 농축된다. 이렇게 농축된 고농도의 젖산은 치아 구성성분인 인산칼슘 중 Ca2+를 녹아내리게 하여 치아 표면 법랑질에 구멍을 내며 치아우식증을 유발하는 것으로 알려져 있다(Sigmund, SS et al., 1992 J. Periodontol. 63:322-331.; Kim et al., Hyunmoonsa 2009 19:307-316). 이에 따라 치아우식증을 예방하기 위하여 스트렙토코커스 뮤탄스에 효과적인 항균제나 GTase의 억제 활성을 보유한 물질을 탐색이 활발하게 진행되고 있다.When Streptococcus mutans comes in contact with sugar, etc. in the oral cavity, polysaccharide crystals are created and stored between the teeth. It attaches to the tooth film and synthesizes insoluble glucan by the action of glucosyltransferase (GTase) enzyme. The synthesized glucan increases the binding force between bacteria and lactic acid formed as metabolic waste accumulates under the polysaccharide crystals and is concentrated. It is known that this concentrated high concentration of lactic acid dissolves Ca 2+ in calcium phosphate, which is a component of teeth, and causes dental caries by making holes in the tooth surface enamel (Sigmund, SS et al., 1992 J. Periodontol. 63 :322-331.; Kim et al., Hyunmoonsa 2009 19:307-316). Accordingly, in order to prevent dental caries, the search for an effective antibacterial agent for Streptococcus mutans or a substance having an inhibitory activity of GTase is being actively conducted.
만성질환인 치아우식증을 치료하기 위하여 일반 항균제를 지속적인 복용할 경우엔 소화기장애, 과민반응, 신장독성, 구강내 독성, 치아변색 등 여러 부작용이 나타날 수 있기 때문에 해당 질환에 특이적이며 안전성이 높은 천연물질을 탐색이 지속적으로 시도되고 있다. 이에 따라 한국등록특허 제1015449390000호에서는 진교 추출물을 유효성분으로 포함하는 치아우식증 예방 또는 치료용 조성물을 개시하고 있으며 한국등록특허 제1015069950000호에서는 오리나루 수피 추출물, 한국등록특허 제1013158050000호에서는 씀바귀 추출물, 한국등록특허 제1012952420000에서는 구맥 추출물을 유효성분으로 포함하는 치아우식증 예방 또는 치료용 조성물을 개시하고 있다. .In the case of continuous use of general antibacterial agents to treat dental caries, a chronic disease, various side effects such as digestive disorders, hypersensitivity reactions, kidney toxicity, oral toxicity, and tooth discoloration may occur. The search for materials is continuously being attempted. Accordingly, Korean Patent No. 1015449390000 discloses a composition for preventing or treating dental caries comprising Jingyo extract as an active ingredient, Korean Patent No. 1015069950000, Orinaru bark extract, Korean Patent No. 1013158050000, Japanese cypress extract, Korean Patent Registration No. 1012952420000 discloses a composition for preventing or treating dental caries, which includes a gourd extract as an active ingredient. .
본 발명은 치우우식증의 원인균인 스트렙토코커스 뮤탄스에 대한 측백나무 잎 추출물 등의 항균 활성 등을 개시한다.The present invention discloses antibacterial activity, such as arborvitae leaf extract, against Streptococcus mutans, a causative agent of dental caries.
본 발명의 목적은 측백나무 잎 추출물 등을 이용한 스트렙토코커스 뮤탄스에 대한 항균 조성물을 제공하는 데 있다.It is an object of the present invention to provide an antibacterial composition against Streptococcus mutans using an extract of arborvitae leaf and the like.
본 발명의 다른 목적은 측백나무 잎 추출물 등을 이용한 구취 제거용 조성물을 제공하는 데 있다.Another object of the present invention is to provide a composition for removing bad breath using an extract of arborvitae leaf and the like.
본 발명의 다른 목적이나 구체적인 목적은 이하에서 제시될 것이다.Other objects or specific objects of the present invention will be set forth below.
본 발명은 아래의 실시예 및 실험예에서 확인되는 바와 같이, 한천배지확산법(disc-agar plate diffusion method)에 의해, 측백나무 잎 추출물과 비파나무 잎 추출물 그리고 이들의 혼합물이 치아우식증의 원인균인 스트렙토코쿠스 뮤탄스(S. mutans)에 항균 활성을 가지고 또 기타 유해 미생물인 대장균(E. coli), 살모넬라 엔테리카(S. enterica), 비브리오 파라해모리티쿠스(V. parahaemolyticus), 바실러스 세레우스(B. cereus)에 대해 항균 활성을 가지며, 또한 구강 청결 항균 효과, 구취 제거 효과 등을 가질 뿐만 아니라 자극도 낮고 향미 등 기호도가 우수함을 확인함으로서 완성된 것이다. As confirmed in the Examples and Experimental Examples below, the present invention is, by the disc-agar plate diffusion method, a arborvitae leaf extract, a loquat leaf extract, and a mixture thereof, which is a causative agent of dental caries, streptococcal. Coccus mutans ( S. mutans ) with antibacterial activity and other harmful microorganisms E. coli ( E. coli ), Salmonella enterica ( S. enterica ), Vibrio para haemolyticus ( V. parahaemolyticus ), Bacillus cereus ( B. cereus ) It has antibacterial activity, and also has an antibacterial effect for oral hygiene, an effect of removing bad breath, etc., and is completed by confirming that the irritation is low and the preference such as flavor is excellent.
전술한 바를 고려할 때, 본 발명은 일 측면에 있어서, 측백나무 잎 추출물, 비파나무 잎 추출물 또는 이들의 혼합물을 유효성분으로 포함하는 스트렙토코쿠스 뮤탄스에 대한 항균용 조성물이나 치아우식증 개선용 조성물로 파악할 수 있고, 다른 측면에 있어서는 측백나무 잎 추출물, 비파나무 잎 추출물 또는 이들의 혼합물을 유효성분으로 포함하는 구취 제거제 조성물로 파악할 수 있으며, 또 다른 측면에 있어서는 측백나무 잎 추출물, 비파나무 잎 추출물 또는 이들의 혼합물을 유효성분으로 포함하는 스트렙토코커스 뮤탄스, 대장균, 살모넬라 엔테리카, 비브리오 파라해모리티쿠스, 바실러스 세레우스에 대한 항균용 조성물로 파악할 수 있다.Considering the above, in one aspect, the present invention provides an antibacterial composition for Streptococcus mutans or a composition for improving dental caries comprising an arborvitae leaf extract, a loquat leaf extract, or a mixture thereof as an active ingredient. In another aspect, it can be identified as a bad breath remover composition comprising an arborvitae leaf extract, a loquat leaf extract or a mixture thereof as an active ingredient, and in another aspect, an arborvitae leaf extract, a loquat leaf extract or It can be identified as an antibacterial composition for Streptococcus mutans, Escherichia coli, Salmonella enterica, Vibrio parahaemolyticus, and Bacillus cereus containing a mixture thereof as an active ingredient.
본 명세서에서, "추출물"이란 추출 대상을 물, 탄소수 1 내지 4의 저급 알콜(메탄올, 에탄올, 부탄올 등), 메틸렌클로라이드, 에틸렌, 아세톤, 헥산, 에테르, 클로로포름, 에틸아세테이트, 부틸아세테이트, N,N-디메틸포름아미드(DMF), 디메틸설폭사이드(DMSO), 1,3-부틸렌글리콜, 프로필렌글리콜 또는 이들의 혼합 용매를 사용하여 침출하여 얻어진 추출물, 이산화탄소, 펜탄 등 초임계 추출 용매를 사용하여 얻어진 추출물, 그 추출물을 분획하여 얻어진 분획물 또는 정유 추출물을 의미하며, 추출 방법은 활성물질의 극성, 추출 정도, 보존 정도를 고려하여 냉침, 환류, 가온, 초음파 방사, 초임계 추출 등 임의의 방법을 적용할 수 있다. 분획된 추출물의 경우 추출물을 특정 용매에 현탁시킨 후 극성이 다른 용매와 혼합·정치시켜 얻은 분획물, 상기 조추출물을 실리카겔 등이 충진된 칼럼에 흡착시킨 후 소수성 용매, 친수성 용매 또는 이들의 혼합 용매를 이동상으로 하여 얻은 분획물을 포함하는 의미이다. 또한 상기 추출물의 의미에는 동결건조, 진공건조, 열풍건조, 분무건조 등의 방식으로 추출 용매가 제거된 농축된 액상의 추출물 또는 고형상의 추출물이 포함된다. 바람직하게는 추출용매로서 물, 에탄올 또는 이들의 혼합 용매, 특히 60% 내지 90%의 에탄올 수용액으로 추출하여 얻어진 것이나 정유 추출물을 의미한다. 일반적으로 정유 추출물은 휘발 성분을 응측시켜 얻어진 오일상의 추출물을 말한다. 정유 추출물의 제조에는 수증기 증류법, 휘발성 용제(벤젠, 헥산 등)를 이용한 용제 추출법, 초임계추출법 등 당업계에 공지된 방법을 이용할 수 있다. As used herein, the term "extract" refers to water, lower alcohols having 1 to 4 carbon atoms (methanol, ethanol, butanol, etc.), methylene chloride, ethylene, acetone, hexane, ether, chloroform, ethyl acetate, butyl acetate, N, Extracts obtained by leaching using N-dimethylformamide (DMF), dimethyl sulfoxide (DMSO), 1,3-butylene glycol, propylene glycol or a mixed solvent thereof, carbon dioxide, pentane, etc. It means an extract obtained, a fraction obtained by fractionating the extract, or an essential oil extract, and the extraction method is an arbitrary method such as chilling, reflux, heating, ultrasonic radiation, supercritical extraction, etc. can be applied. In the case of a fractionated extract, a fraction obtained by suspending the extract in a specific solvent and mixing and standing still with a solvent having a different polarity, and adsorbing the crude extract to a column filled with silica gel, etc. It is meant to include the fraction obtained as a mobile phase. In addition, the meaning of the extract includes a concentrated liquid extract or solid extract from which the extraction solvent is removed by freeze drying, vacuum drying, hot air drying, spray drying, or the like. Preferably, it refers to an essential oil extract or obtained by extraction with water, ethanol, or a mixed solvent thereof as an extraction solvent, particularly 60% to 90% of an aqueous ethanol solution. In general, an essential oil extract refers to an oily extract obtained by condensing a volatile component. For the preparation of the essential oil extract, methods known in the art, such as steam distillation, solvent extraction using a volatile solvent (benzene, hexane, etc.), and supercritical extraction may be used.
또 본 명세서에서 "유효성분"이란 단독으로 목적하는 활성을 나타내거나 또는 그 자체는 활성이 없는 담체와 함께 활성을 나타낼 수 있는 성분을 의미한다.In addition, as used herein, the term "active ingredient" refers to a component capable of exhibiting the desired activity alone or in combination with a carrier that has no activity by itself.
또 본 명세서에서, "치아 우식증 개선"이란 대상 질환인 치아우식증의 예방, 치료 또는 증상의 경감을 포함하는 의미이다. In addition, in the present specification, "improving dental caries" means preventing, treating, or alleviating symptoms of dental caries, which is a target disease.
또 본 명세서에서, "항균"은 해당 미생물의 사멸 유도 또는 증식 억제를 포함하는 의미이다.Also, in the present specification, "antibacterial" is meant to include induction of death or inhibition of proliferation of the corresponding microorganism.
본 발명의 항균용 조성물이나 치주우식증 개선용 조성물(이하 통칭하여 "본 발명의 조성물")은 그 유효성분을 용도, 제형, 배합 목적 등에 따라 의도하는 항균 활성, 치아우식증 개선 활성 등을 나타낼 수 있는 한 임의의 양(유효량)으로 포함할 수 있는데, 통상적인 유효량은 조성물 전체 중량을 기준으로 할 때 0.001 중량 % 내지 15 중량 % 범위 내에서 결정될 것이다. 여기서 "유효량"이란 그 적용 대상인 포유동물 바람직하게는 사람에게 의료 전문가 등의 제언에 의한 투여 기간 동안 본 발명의 조성물이 투여될 때, 치아우식증 등의 개선 등의 의도한 의료적·약리학적 효과를 나타낼 수 있는 유효성분의 양을 말한다. 이러한 유효량은 당업자의 통상의 능력 범위 내에서 실험적으로 결정될 수 있다.The antibacterial composition of the present invention or the composition for improving periodontal caries (hereinafter collectively referred to as the "composition of the present invention") may exhibit the intended antibacterial activity, dental caries improvement activity, etc. It may be included in any amount (effective amount) as long as the effective amount will be determined within the range of 0.001% to 15% by weight based on the total weight of the composition. Herein, the term "effective amount" refers to the intended medical and pharmacological effects such as improvement of dental caries, etc., when the composition of the present invention is administered to a mammal, preferably a human, to which it is applied during the administration period as suggested by a medical professional. It refers to the amount of active ingredient that can be displayed. Such effective amounts can be determined empirically within the ordinary ability of one of ordinary skill in the art.
본 발명의 조성물은 구체적인 양태에 있어서, 식품 조성물로서 파악할 수 있다.In a specific aspect, the composition of this invention can be grasped|ascertained as a food composition.
본 발명의 식품 조성물은 어떠한 형태로도 제조될 수 있으며, 예컨대 차, 쥬스, 탄산음료, 이온음료 등의 음료류, 우유, 요구루트 등의 가공 유류, 껌류, 떡, 한과, 빵, 과자, 면 등의 식품류, 정제, 캡슐, 환, 과립, 액상, 분말, 편상, 페이스트상, 시럽, 겔, 젤리, 바 등의 건강기능식품 제제류 등으로 제조될 수 있다. 또 본 발명의 식품 조성물은 법률상·기능상의 구분에 있어서 제조·유통 시점의 시행 법규에 부합하는 한 임의의 제품 구분을 띨 수 있다. 예컨대 건강기능식품에관한법률에 따른 건강기능식품이거나, 식품위생법의 식품공전(식약처 고시, 식품의 기준 및 규격)상 각 식품유형에 따른 과자류, 두류, 두유류, 발효음료류, 특수용도식품 등일 수 있다.The food composition of the present invention may be prepared in any form, for example, beverages such as tea, juice, carbonated beverages, ionic beverages, processed oils such as milk and yogurt, gums, rice cakes, Korean sweets, bread, confectionery, noodles, etc. of foods, tablets, capsules, pills, granules, liquids, powders, flakes, pastes, syrups, gels, jellies, bars and the like health functional food preparations. In addition, the food composition of the present invention may have any product classification in terms of legal and functional classification as long as it conforms to the enforcement laws at the time of manufacture and distribution. For example, it is a health functional food according to the Health Functional Food Act, or confectionery, beans, soy milk, fermented beverages, special purpose food, etc. can
본 발명의 식품 조성물에는 그 유효성분 이외에 식품첨가물이 포함될 수 있다. 식품첨가물은 일반적으로 식품을 제조, 가공 또는 보존함에 있어 식품에 첨가되어 혼합되거나 침윤되는 물질로서 이해되는데, 식품과 함께 매일 그리고 장기간 복용되므로 그 안전성이 보장되어야 한다. 식품위생법에 따른 식품첨가물공전(식약처 고시, 식품첨가물 기준 및 규격)에는 안전성이 보장된 식품첨가물이 화학적 합성품, 천연 첨가물, 혼합 제제류로 구분하여 한정적으로 규정되어 있다. The food composition of the present invention may contain food additives in addition to the active ingredients thereof. Food additives are generally understood as substances that are added to, mixed or infiltrated with food in manufacturing, processing, or preserving food. The Food Additives Ordinance (Ministry of Food and Drug Safety Notice, Food Additive Standards and Specifications) according to the Food Sanitation Act restricts food additives with guaranteed safety by classifying them into chemically synthesized products, natural additives, and mixed preparations.
이들 식품첨가물은 기능적 측면에 있어서는 감미제, 풍미제, 보존제, 유화제, 산미료, 점증제 등으로 구분될 수 있다. These food additives may be classified into sweeteners, flavoring agents, preservatives, emulsifiers, acidulants, thickeners, and the like in terms of functionality.
감미제는 식품에 적당한 단맛을 부여하기 위하여 사용되는 것으로, 천연의 것이거나 합성된 것을 사용할 수 있다. 바람직하게는 천연 감미제를 사용하는 경우인데, 천연 감미제로서는 옥수수 시럽 고형물, 꿀, 수크로오스, 프룩토오스, 락토오스, 말토오스 등의 당 감미제를 들 수 있다. The sweetener is used to impart appropriate sweetness to food, and natural or synthetic ones may be used. Preferably, a natural sweetener is used. Examples of the natural sweetener include sugar sweeteners such as corn syrup solids, honey, sucrose, fructose, lactose, and maltose.
풍미제는 맛이나 향을 좋게 하기 위하여 사용될 수 있는데, 천연의 것과 합성된 것 모두 사용될 수 있다. 바람직하게는 천연의 것을 사용하는 경우이다. 천연의 것을 사용할 경우에 풍미 이외에 영양 강화의 목적도 병행할 수 있다. 천연 풍미제로서는 사과, 레몬, 감귤, 포도, 딸기, 복숭아 등에서 얻어진 것이거나 녹차잎, 둥굴레, 대잎, 계피, 국화 잎, 자스민 등에서 얻어진 것일 수 있다. 또 인삼(홍삼), 죽순, 알로에 베라, 은행 등에서 얻어진 것을 사용할 수 있다. 천연 풍미제는 액상의 농축액이나 고형상의 추출물일 수 있다. 경우에 따라서 합성 풍미제가 사용될 수 있는데, 합성 풍미제는 에스테르, 알콜, 알데하이드, 테르펜 등이 이용될 수 있다. Flavoring agents may be used to improve taste or aroma, and both natural and synthetic ones may be used. Preferably, it is a case where a natural thing is used. In the case of using a natural one, the purpose of nutritional enhancement in addition to flavor may be concurrently used. As a natural flavoring agent, it may be obtained from apple, lemon, tangerine, grape, strawberry, peach, etc., or it may be obtained from green tea leaf, dandelion, bamboo leaf, cinnamon, chrysanthemum leaf, jasmine, etc. In addition, those obtained from ginseng (red ginseng), bamboo shoots, aloe vera, ginkgo biloba, etc. can be used. The natural flavoring agent may be a liquid concentrate or a solid extract. Optionally, a synthetic flavoring agent may be used, and the synthetic flavoring agent may be an ester, an alcohol, an aldehyde, a terpene, or the like.
보존제로서는 소듐 소르브산칼슘, 소르브산나트륨, 소르브산칼륨, 벤조산칼슘, 벤조산나트륨, 벤조산칼륨, EDTA(에틸렌디아민테트라아세트산) 등이 사용될 수 있고, 또 유화제로서는 아카시아검, 카르복시메틸셀룰로스, 잔탄검, 펙틴 등을 들 수 있으며, 산미료로서는 연산, 말산, 푸마르산, 아디프산, 인산, 글루콘산, 타르타르산, 아스코르브산, 아세트산, 인산 등이 사용될 수 있다. 산미료는 맛을 증진시키는 목적 이외에 미생물의 증식을 억제할 목적으로 식품 조성물이 적정 산도로 되도록 첨가될 수 있다.As a preservative, sodium calcium sorbate, sodium sorbate, potassium sorbate, calcium benzoate, sodium benzoate, potassium benzoate, EDTA (ethylenediaminetetraacetic acid), etc. can be used, and as an emulsifier, acacia gum, carboxymethylcellulose, xanthan gum, Pectin etc. are mentioned, and acidulant, malic acid, fumaric acid, adipic acid, phosphoric acid, gluconic acid, tartaric acid, ascorbic acid, acetic acid, phosphoric acid, etc. can be used as an acidulant. Acidulant may be added so that the food composition has an appropriate acidity for the purpose of inhibiting the growth of microorganisms in addition to the purpose of enhancing the taste.
점증제로서는 현탁화 구현제, 침강제, 겔형성제, 팽화제 등이 사용될 수 있다.As the thickening agent, a suspending agent, a settling agent, a gel-forming agent, a bulking agent, and the like can be used.
본 발명의 식품 조성물은 전술한 바의 식품첨가물 이외에, 기능성과 영양성을 보충, 보강할 목적으로 당업계에 공지되고 식품첨가물로서 안정성이 보장된 생리활성 물질이나 미네랄류를 포함할 수 있다.The food composition of the present invention may contain, in addition to the food additives as described above, physiologically active substances or minerals known in the art for the purpose of supplementing and reinforcing functionality and nutrition and guaranteed stability as food additives.
그러한 생리활성 물질로서는 녹차 등에 포함된 카테킨류, 비타민 B1, 비타민 C, 비타민 E, 비타민 B12 등의 비타민류, 토코페롤, 디벤조일티아민 등을 들 수 있으며, 미네랄류로서는 구연산 칼슘 등의 칼슘 제제, 스테아린산마그네슘 등의 마그네슘 제제, 구연산철 등의 철 제제, 염화 크롬, 요오드칼륨, 셀레늄, 게르마늄, 바나듐, 아연 등을 들 수 있다. Examples of such physiologically active substances include catechins contained in green tea and the like, vitamins such as vitamin B1, vitamin C, vitamin E, and vitamin B12, tocopherol, dibenzoylthiamine, and the like. Magnesium preparations, such as iron preparations, such as iron citrate, chromium chloride, potassium iodide, selenium, germanium, vanadium, zinc, etc. are mentioned.
본 발명의 식품 조성물에는 전술한 바의 식품첨가물이 제품 유형에 따라 그 첨가 목적을 달성할 수 있는 적량으로 포함될 수 있다.In the food composition of the present invention, the food additives as described above may be included in an appropriate amount to achieve the purpose of the addition according to the type of product.
본 발명의 식품 조성물에 포함될 수 있는 기타의 식품첨가물과 관련하여서는 식품공전이나 식품첨가물 공전을 참조할 수 있다.In relation to other food additives that may be included in the food composition of the present invention, reference may be made to the Food Codex or the Food Additives Codex.
본 발명의 조성물은 다른 구체적인 양태에 있어서는 약제학적 조성물로 파악될 수 있다.The composition of the present invention may be identified as a pharmaceutical composition in another specific embodiment.
본 발명의 약제학적 조성물은 유효성분 이외에 약제학적으로 허용되는 담체를 포함하여 당업계에 공지된 통상의 방법으로 구강청결제 조성물 제형 또는 치약 조성물 제형 등의 의약외품 제형, 경구용 제형 또는 비경구용 제형 등의 의약품 제형으로 제조될 수 있다. 여기서 "약제학적으로 허용되는" 의미는 유효성분의 활성을 억제하지 않으면서 적용(처방) 대상이 적응 가능한 이상의 독성을 지니지 않는다는 의미이다.The pharmaceutical composition of the present invention includes a pharmaceutically acceptable carrier in addition to the active ingredient by a conventional method known in the art, such as a mouthwash composition formulation or a quasi-drug formulation such as a toothpaste formulation, an oral formulation, or a parenteral formulation. It can be prepared in pharmaceutical formulations. Here, "pharmaceutically acceptable" means that it does not inhibit the activity of an active ingredient and does not have toxicity beyond what the application (prescription) target can adapt.
본 발명의 조성물이 구강청결제 조성물 제형, 치약 조성물 제형으로 제조될 경우, 페이스트상, 분말상, 분무상 또는 액상(가글상)으로 제조될 수 있으며, 본 발명의 유효성분인 카르노스산을 포함하는 이외에, 충치 예방 약효 성분, 연마제 성분, 습윤제 성분, 결합제 성분, 기포제 성분, 감미제 성분, 향료 성분 및 색소 성분으로 이루어진 군에서 선택된 어느 하나 이상의 성분을 추가로 포함할 수 있다.When the composition of the present invention is prepared as a mouthwash composition formulation or a toothpaste composition formulation, it may be prepared in a paste form, powder form, spray form or liquid (gargle form), and in addition to containing carnosic acid as the active ingredient of the present invention, It may further include any one or more components selected from the group consisting of an anti-cavity drug component, an abrasive component, a wetting agent component, a binder component, a foaming component, a sweetener component, a fragrance component, and a color component.
상기 충치 예방 약효 성분으로서 불화나트륨, 불화인산나트륨, 불화아민, 불화주석, 클로로헥시딘, 세틸피리디움 클로라이드, 트리클로산, 자일리톨, 죽염 및 프로폴리스으로 이루어진 군에서 선택되는 어느 하나 이상의 성분을 사용할 수 있다.Any one or more ingredients selected from the group consisting of sodium fluoride, sodium fluorophosphate, amine fluoride, tin fluoride, chlorhexidine, cetylpyridium chloride, triclosan, xylitol, bamboo salt, and propolis can be used as the tooth decay prevention medicinal ingredient there is.
또 상기 연마제 성분으로서 침강실리카, 실리카겔, 지르코늄실리케이트, 인산일수소칼슘, 무수인산일수소칼슘, 함수알루미나, 경질탄산칼슘, 중질탄산칼슘, 칼슘피로인산염, 불용성메타인산염 및 알루미늄실리케이트으로 이루어진 군에서 선택되는 어느 하나 이상의 성분을 사용할 수 있다. In addition, the abrasive component is selected from the group consisting of precipitated silica, silica gel, zirconium silicate, calcium monohydrogen phosphate, anhydrous calcium monohydrogen phosphate, hydrous alumina, light calcium carbonate, heavy calcium carbonate, calcium pyrophosphate, insoluble metaphosphate, and aluminum silicate. Any one or more components may be used.
또 상기 습윤제 성분으로서 글리세린, 소르비톨, 자일리톨, 폴리에틸렌글리콜 및 프로필렌글리콜로 이루어진 군에서 선택되는 어느 하나 이상을 사용할 수 있다. 이러한 습윤제 성분은 페이스트 제형을 만드는데 필수적인 베이스 성분으로 치약이 공기 중에 노출될 때 건조, 고화되는 것을 방지하고 치약의 표면에 윤기를 제공할 뿐만 아니라, 종류에 따라서는 양치시 감미 효과를 주는 역할을 한다.In addition, as the humectant component, any one or more selected from the group consisting of glycerin, sorbitol, xylitol, polyethylene glycol and propylene glycol may be used. This humectant component is an essential base component for making a paste formulation. When the toothpaste is exposed to the air, it prevents drying and solidification, and provides gloss to the surface of the toothpaste. Depending on the type, it plays a role in giving a sweetening effect when brushing teeth. .
또 상기 결합제 성분으로서 수용성의 고분자 종류라면 어떤 것이든 사용할 수 있는데, 바람직하게는 카르복시메틸셀룰로오스나트륨, 카라기난류, 잔탄검 등을 사용할 수 있다. 상기 결합제는 고체인 분말 성분과 액체 성분이 분리되지 않게 하는 작용을 한다.In addition, as the binder component, any water-soluble polymer may be used. Preferably, sodium carboxymethyl cellulose, carrageenans, xanthan gum, etc. may be used. The binder acts to prevent the solid powder component from being separated from the liquid component.
또 상기 기포제 성분으로서는 음이온성 계면활성제인 라우릴황산나트륨을 사용하는 것이 바람직하며, 제형의 특성에 따라 보조적으로 폴리옥시에틸렌폴리옥시프로필렌의 공중합체(폴록사머), 폴리옥시에틸렌경화피마자유 또는 폴리옥시에틸렌솔비탄 지방산에스테르 등의 비이온성 계면활성제를 사용할 수 있다. 이러한 기포제 성분은 제품의 사용감을 증진시키고 세정 작용을 도와주며, 기타 약효 성분의 분산 및 침투를 신속하게 하고, 계면장력을 감소시킴으로서 구강내 이물질을 쉽게 떨어지게 하는 작용을 한다.In addition, as the foaming agent component, it is preferable to use sodium lauryl sulfate, an anionic surfactant, and depending on the characteristics of the formulation, a copolymer of polyoxyethylene polyoxypropylene (poloxamer), polyoxyethylene hydrogenated castor oil or polyoxy Nonionic surfactants, such as ethylene sorbitan fatty acid ester, can be used. These foaming ingredients enhance the feeling of use of the product, help the cleaning action, speed up the dispersion and penetration of other active ingredients, and reduce interfacial tension, thereby allowing foreign substances in the oral cavity to fall off easily.
또 상기 향료 성분, 감미제 성분, 색소 성분 등은 치약 조성물의 사용감을 좋게 하기 위한 것으로, 향료 성분으로서는 식용 가능한 것을 사용하는 것이 바람직한데, 그러한 성분으로서는 예컨대 멘톨, 페파민트 오일, 스피아민트 오일, 세이지, 유칼립톨, 메틸살리실레이트 또는 과일추출물 등을 들 수 있다.In addition, the perfume component, sweetener component, color component, etc. are for improving the feeling of use of the toothpaste composition, and it is preferable to use an edible component as the flavor component. As such components, for example, menthol, peppermint oil, spearmint oil, sage, eucal lyptol, methyl salicylate, or fruit extract.
또 상기 감미제 성분으로서는 삭카린나트륨을 사용하는 것이 바람직하며, 상기 색소 성분으로서는 식용색소를 사용하는 것이 바람직하다.In addition, it is preferable to use sodium saccharin as the sweetener component, and it is preferable to use food coloring as the pigment component.
본 발명의 약제학적 조성물이 경구용 제형으로 제조될 경우, 적합한 담체와 함께 당업계에 공지된 방법에 따라 분말, 과립, 정제, 환제, 당의정제, 캡슐제, 액제, 겔제, 시럽제, 현탁액, 웨이퍼 등의 제형으로 제조될 수 있다. 이때 약제학적으로 허용되는 적합한 담체의 예로서는 락토스, 글루코스, 슈크로스, 덱스트로스, 솔비톨, 만니톨, 자일리톨 등의 당류, 옥수수 전분, 감자 전분, 밀 전분 등의 전분류, 셀룰로오스, 메틸셀룰로오스, 에틸셀룰로오스, 나트륨 카르복시메틸셀룰로오스, 하이드록시프로필메틸셀룰로오스 등의 셀룰로오스류, 폴리비닐 피롤리돈, 물, 메틸히드록시벤조에이트, 프로필히드록시벤조에이트, 마그네슘 스테아레이트, 광물유, 맥아, 젤라틴, 탈크, 폴리올, 식물성유 등을 들 수 있다. 제제화활 경우 필요에 따라 충진제, 증량제, 결합제, 습윤제, 붕해제, 계면활성제 등의 희석제 및/또는 부형제를 포함하여 제제화할 수 있다.When the pharmaceutical composition of the present invention is prepared as an oral dosage form, powders, granules, tablets, pills, dragees, capsules, liquids, gels, syrups, suspensions, wafers, together with a suitable carrier according to methods known in the art. It can be prepared in a formulation such as In this case, examples of suitable pharmaceutically acceptable carriers include sugars such as lactose, glucose, sucrose, dextrose, sorbitol, mannitol, and xylitol, starches such as corn starch, potato starch, wheat starch, cellulose, methylcellulose, ethylcellulose, Cellulose such as sodium carboxymethylcellulose and hydroxypropylmethylcellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, magnesium stearate, mineral oil, malt, gelatin, talc, polyol, vegetable and oil. In the case of formulation activity, the formulation may be formulated by including diluents and/or excipients such as fillers, extenders, binders, wetting agents, disintegrants, and surfactants as necessary.
본 발명의 약제학적 조성물이 비경구용 제형으로 제조될 경우, 적합한 담체와 함께 당업계에 공지된 방법에 따라 주사제, 경피 투여제, 비강 흡입제 및 좌제의 형태로 제제화될 수 있다. 주사제로 제제화활 경우 적합한 담체로서는 멸균수, 에탄올, 글리세롤이나 프로필렌 글리콜 등의 폴리올 또는 이들의 혼합물을 들수 있으며, 바람직하게는 링거 용액, 트리에탄올 아민이 함유된 PBS(phosphate buffered saline)나 주사용 멸균수, 5% 덱스트로스 같은 등장 용액 등을 사용할 수 있다. 경피 투여제로 제제화할 경우 연고제, 크림제, 로션제, 겔제, 외용액제, 파스타제, 리니멘트제, 에어롤제 등의 형태로 제제화될 수 있다. 비강 흡입제의 경우 디클로로플루오로메탄, 트리클로로플루오로메탄, 디클로로테트라플루오로에탄, 이산화탄소 등의 적합한 추진제를 사용하여 에어로졸 스프레이 형태로 제제화될 수 있으며, 좌제의 기제로는 위텝솔(witepsol), 트윈(tween) 61, 폴리에틸렌글리콜류, 카카오지, 라우린지, 폴리옥시에틸렌 소르비탄 지방산 에스테르류, 폴리옥시에틸렌 스테아레이트류, 소르비탄 지방산 에스테르류 등이 사용될 수 있다.When the pharmaceutical composition of the present invention is prepared for parenteral use, it may be formulated in the form of injections, transdermal administrations, nasal inhalants and suppositories together with suitable carriers according to methods known in the art. In the case of formulation for injection, suitable carriers include sterile water, ethanol, polyols such as glycerol or propylene glycol, or mixtures thereof, preferably Ringer's solution, PBS (phosphate buffered saline) containing triethanolamine, or sterile water for injection. , an isotonic solution such as 5% dextrose may be used. When formulated for transdermal administration, it may be formulated in the form of an ointment, a cream, a lotion, a gel, an external solution, a pasta agent, a liniment agent, an air roll, and the like. In the case of nasal inhalants, it can be formulated in the form of an aerosol spray using a suitable propellant such as dichlorofluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide, etc. (tween) 61, polyethylene glycols, cacao fat, laurin fat, polyoxyethylene sorbitan fatty acid esters, polyoxyethylene stearate, sorbitan fatty acid esters, and the like may be used.
약제학적 조성물의 제제화와 관련하여서는 당업계에 공지되어 있으며, 보다 구체적인 사항은 문헌[Remington's Pharmaceutical Sciences(19th ed., 1995)] 등을 참조할 수 있다. 상기 문헌은 본 명세서의 일부로서 간주 된다.Formulation of pharmaceutical compositions is known in the art, and for more specific details, reference may be made to the literature [Remington's Pharmaceutical Sciences (19th ed., 1995)] and the like. This document is considered a part of this specification.
본 발명의 약제학적 조성물의 바람직한 투여량은 환자의 상태, 체중, 성별, 연령, 환자의 중증도, 투여 경로에 따라 1일 0.001mg/kg ~ 10g/kg 범위, 바람직하게는 0.001mg/kg ~ 1g/kg 범위일 수 있다. 투여는 1일 1회 또는 수회로 나누어 이루어질 수 있다. 이러한 투여량은 어떠한 측면으로든 본 발명의 범위를 제한하는 것으로 해석되지 아니 한다. A preferred dosage of the pharmaceutical composition of the present invention is in the range of 0.001 mg/kg to 10 g/kg per day, preferably 0.001 mg/kg to 1 g, depending on the patient's condition, weight, sex, age, patient's severity, and administration route. It can be in the range /kg. Administration may be performed once or divided into several times a day. Such dosages are not to be construed as limiting the scope of the invention in any respect.
전술한 바와 같이, 본 발명에 따르면 측백나무 잎 등을 이용한 스트렙토코커스 뮤탄스 균에 대한 항균 조성물을 제공할 수 있다. 본 발명의 조성물은 구강 청결제, 치약 등의 의약외품, 기능성 식품, 약품 등으로 제품화될 수 있다.As described above, according to the present invention, it is possible to provide an antibacterial composition against Streptococcus mutans using arborvitae leaves and the like. The composition of the present invention may be commercialized as a mouthwash, quasi-drugs such as toothpaste, functional food, medicine, and the like.
도 1 내지 도 5는 항균 활성 결과이다.
도 6은 세균수 측정 결과이다.
도 7은 구강내 구취가스 분석 결과이다.1 to 5 are results of antibacterial activity.
6 is a result of measuring the number of bacteria.
7 is an analysis result of bad breath gas in the oral cavity.
이하 본 발명을 실시예를 참조하여 설명한다. 그러나 본 발명의 범위가 이러한 실시예에 한정되는 것은 아니다.Hereinafter, the present invention will be described with reference to Examples. However, the scope of the present invention is not limited to these examples.
<실시예> 구강 미생물 등에 대한 항균 활성 실험, 구강내 세균수 측정 실험, 구강내 구취가스 분석 실험 및 관능평가 실험<Example> Antibacterial activity test for oral microorganisms, etc., oral bacterial count test, oral bad breath gas analysis test and sensory evaluation test
1.시료 및 방법1. Samples and Methods
1.1 시료1.1 Samples
에탄올 추출물 시료는, 측백나무(Thuja orientalis, 추출 부위: 잎), 비파나무(Eriobotrya Japonica, 추출 부위: 잎) 각 건조 분말에 10배 중량의 80% 에탄올을 가하고 80℃에서 4시간 동안 추출하고 여과지(Whatman #2)로 여과 후 감압농축하고 동결건조하여 분말로 준비하였다.For the ethanol extract sample , 10 times the weight of 80% ethanol was added to each dried powder of Thuja orientalis ( Thuja orientalis , extraction part: leaf), loquat tree ( Eriobotrya Japonica , extraction part: leaf), and extracted at 80 ° C. for 4 hours, followed by filter paper After filtration with (Whatman #2), it was concentrated under reduced pressure and lyophilized to prepare a powder.
열수 추출물 시료는 측백나무 잎, 비파나무 잎 각 건조 분말을 물과 1:10배 중량을 넣고 환류 냉각기가 장착된 추출기에서 각각 100℃에서 4시간 동안 추출하였다. 그 추출물은 에탄올 추출물시료와 동일한 방법으로 하여 분말을 얻었다. For the hot water extract sample, dry powders of arborvitae leaf and loquat leaf were added 1:10 times with water and extracted at 100° C. for 4 hours in an extractor equipped with a reflux condenser. The extract was obtained in the same manner as the ethanol extract sample to obtain a powder.
정유 추출물 시료는 측백나무 잎, 비파나무 잎 각 건조 분말 각각 100℃에서 4시간 동안 열수에서 증류 추출하여 준비하였다. 추출조건은 SDE(simultaneous distillation and extraction) 장치를 사용하였다. 포집된 정유 속의 함유되어 있는 수분을 제거하기 위해서 황산나트륨을 가하여 여과하였다. The essential oil extract samples were prepared by distillation extraction in hot water at 100° C. for 4 hours, respectively, of dry powders of arborvitae leaf and loquat leaf. As the extraction conditions, a simultaneous distillation and extraction (SDE) apparatus was used. Sodium sulfate was added and filtered to remove moisture contained in the captured essential oil.
혼합물 시료는 상기 에탄올 추출물과 상기 정유 추출물을 동일 중량으로 혼합하여 얻었다. A mixture sample was obtained by mixing the ethanol extract and the essential oil extract in equal weights.
시제품 시료는 아래 표에 기재된 바의 중량%로 준비하였다. 5% 에탄올은 보존제 및 가용화제의 용도로 사용되었으며, 또 구강청결제로서의 사용을 고려하여 계면활성제 성질을 가진 녹두 80% 에탄올 추출물(상기와 동일하게 제조)을 첨가하여 사용하였다. 시제품의 총량 100%를 맞추기 위해서 계면활성제인 Tween 20은 0.0001%, 충치예방에 쓰이는 NaF를 0.05%를 함유한 멸균수를 모든 처리구에 동일하게 적용하여 시제품을 제조하였다. Prototype samples were prepared in % by weight as indicated in the table below. 5% ethanol was used as a preservative and solubilizer, and in consideration of use as a mouthwash, 80% ethanol extract of mung bean (prepared in the same manner as above) with surfactant properties was added and used. In order to meet 100% of the total amount of the prototype, sterile water containing 0.0001% of
정유loquat
essential oil
에탄올 추출물arboretum
ethanol extract
에탄올 추출물loquat
ethanol extract
추출물green gram
extract
1.2 항균 활성 실험1.2 Antibacterial activity test
항균 활성은, 구강 미생물인 스트렙토코쿠스 뮤탄스(streptococcus mutans)와 기타 유해 미생물인 대장균(E. coli), 살모넬라 엔테리카(S. enterica), 비브리오 파라해모리티쿠스(V. parahaemolyticus), 바실러스 세레우스(B. cereus)를 대상으로 하였다. Antibacterial activity is, oral microorganisms Streptococcus mutans ( streptococcus mutans ) and other harmful microorganisms Escherichia coli ( E. coli ), Salmonella enterica ( S. enterica ), Vibrio parahaemolyticus ( V. parahaemolyticus ), Bacillus Cereus ( B. cereus ) was targeted.
배지는, 스트렙토코쿠스 뮤탄스에 대해서는 BHI(Brain Heart Infusion 배지(Bacto BHI, BD, USA)를, 기타 유해 미생물에 대해서는 LB 배지(Bacto-LB, BD, USA)를 사용하였다. As a medium, for Streptococcus mutans, BHI (Brain Heart Infusion medium (Bacto BHI, BD, USA) was used, and for other harmful microorganisms, LB medium (Bacto-LB, BD, USA) was used.
시료의 항균 활성 실험은 한천배지확산법(disc-agar plate diffusion method)을 이용하였다. 구체적으로 시료를 디에틸에테르에 일정한 농도로 녹여 0.45 ㎛ membrane filter(Milipore, USA)로 여과하여 멸균된 filter paper disc(Toyo seisakusho, 8 ㎜)에 다양한 농도로 40 ㎕씩 흡수시킨 후 질소가스를 이용하여 에테르를 제거하고 시험용 평판배지 위에 놓고 28℃의 incubator에서 20시간 배양한 다음 disc 주변의 clear zone의 직경(㎜)을 측정하였다. Antibacterial activity of the sample was tested using the agar medium diffusion method (disc-agar plate diffusion method). Specifically, the sample was dissolved in diethyl ether at a constant concentration, filtered with a 0.45 μm membrane filter (Milipore, USA), and absorbed into a sterilized filter paper disc (Toyo seisakusho, 8 mm) by 40 μl at various concentrations, followed by nitrogen gas. After removing the ether, it was placed on a test plate medium and incubated for 20 hours in an incubator at 28°C, and then the diameter (mm) of the clear zone around the disc was measured.
1.3 시제품 세균수 측정 실험1.3 Prototype bacteria count test
시제품에 대한 유해미생물들의 항균 활성 실험은, 가글액에 시험균액을 첨가하여 LB 배지(기타 유해균) 및 BHI(Brain Heart Infusion) 배지(뮤탄스 균)를 이용하여 세균수를 측정하여 수행하였다. The antibacterial activity experiment of harmful microorganisms on the prototype was performed by measuring the number of bacteria using LB medium (other harmful bacteria) and BHI (Brain Heart Infusion) medium (bacterium mutans) by adding the test bacteria solution to the gargle solution.
시험균액은 S. mutans와 E. coli, S. enterica, V. parahaemolyticus, B. cereus 혼합균주를 사용하였고 생균수가 1-9.9×104 CFU/ml이 되도록 멸균생리식염수에 희석하여 시험균액으로 사용하였다.A mixed strain of S. mutans , E. coli, S. enterica, V. parahaemolyticus, and B. cereus was used for the test solution, and diluted with sterile physiological saline so that the number of viable cells was 1-9.9 × 10 4 CFU/ml was used as the test solution. did.
가글원액 20 ml에 시험균액 0.2 ml을 첨가하여 0, 30, 60, 90, 120초간 방치하였다. 방치된 균들을 배양하기 위하여 45-50℃의 액체화된 LB 및 BHI배지에 처리완료된 시료 각 1 ml씩 취하여 액체화된 배지에 넣고 petri-dish에 넣어 응고시켰다. 응고된 petri-dish는 35℃, 충치균은 48시간, 식품유해균은 24시간동안 배양하여 자라난 균주의 밀도를 측정하였다.0.2 ml of the test bacteria solution was added to 20 ml of the gargle stock solution and left for 0, 30, 60, 90, and 120 seconds. In order to culture the left bacteria, 1 ml of each of the treated samples was taken in liquefied LB and BHI medium at 45-50° C., put in the liquid medium, and put in petri-dish to coagulate. The coagulated petri-dish was cultured at 35°C, tooth decay bacteria for 48 hours, and food harmful bacteria for 24 hours to measure the density of the grown strains.
1.4 구강내 구취가스 분석 실험1.4 Oral bad breath gas analysis test
동신대학교 대학생(남 3명, 여자 7명) 총 10명을 대상으로 점심식사 후 30분동안 음식물을 섭취하지 못하게 하였다. 시료를 채취하기 위해 실험 1분전에는 입을 다물고 코로 호흡하게 만든 다음 500 ml 부피의 공기주머니(폴리프로필렌 재질)를 이용하여 입속의 공기를 2분동안 서서히 가득 채우게 하였다. 다음 시제품 및 린스테린의 가글액 20 ml을 1분간 입안을 행구고 시료를 뱉어낸 다음 입을 다물고 각각 10분, 20분, 30분 단위로 구취가스를 입으로 2분간 서서히 불어 가득채운다음 밀봉하였다. 채취된 공기주머니는 25℃ 실온에 보관하면서 분석하였다. A total of 10 Dongshin University college students (3 males, 7 females) were prohibited from eating for 30 minutes after lunch. To collect the sample, one minute before the experiment, the mouth was closed and the nose was made to breathe, and then the air in the mouth was gradually filled for 2 minutes using an air bag (polypropylene material) with a volume of 500 ml. Next, rinse the mouth with 20 ml of the gargle solution of the prototype and linsterin for 1 minute, spit out the sample, close the mouth, and slowly fill the mouth with bad breath gas for 2 minutes at intervals of 10, 20, and 30 minutes, respectively, and then seal. The collected air bags were analyzed while being stored at room temperature at 25°C.
채취된 입속의 구취가스는 가스검지기(GV-100S, Gastec, Japan)를 사용하여 methylmercaptane성분을 정량하였다. 정량 방법은 멸균수로 입안을 행굼처리한 대조구 측정값과 시제품처리 측정값을 상호비교(구취억제율(%) = 100 x (최종값 - 최초값)/최초값)하여 수행하였다. The methylmercaptane component was quantified using a gas detector (GV-100S, Gastec, Japan) for the collected bad breath gas. The quantitative method was performed by comparing the measured value of the control group treated with sterile water and the measured value of the prototype treatment (bad breath inhibition rate (%) = 100 x (final value - initial value)/initial value).
1.5 관능평가1.5 Sensory evaluation
관능평가는 동신대학교 학생 20명을 관능평가 요원으로 하여 자극도(irritation degree), 색상, 맛, 구강 촉감, 냄새, 기호도에 대해서 아래의 5점 척도법에 따라 수행하고 각 평가점수를 합산하였다. 비교예(Control)는 시중에 판매되는 리스테린(녹차 첨가)를 가지고 수행하였다. For sensory evaluation, 20 Dongshin University students were used as sensory evaluation agents for irritation degree, color, taste, mouth feel, smell, and preference according to the following 5-point scale method, and each evaluation score was summed. Comparative Example (Control) was carried out with commercially available Listerine (with green tea added).
2. 실험 결과2. Experimental results
2.1 항균 활성 실험 결과2.1 Antibacterial activity test result
뮤탄스 균에 대한 항균 활성 실험 결과를, 혼합물에 대해서는 도 1에 나타내었고, 시제품에 대해서는 도 2에 나타내었다. The results of the antimicrobial activity test against mutans bacteria were shown in FIG. 1 for the mixture, and shown in FIG. 2 for the prototype.
도 1을 참조하여 보면, 혼합물 시료가 에탄올 추출물의 혼합물이든 정유 추출물의 혼합물이든 각각 단독 시료보다 항균 활성이 높음을 알 수 있으며 시제품 시료의 경우 시료 첨가량이 증가할 수록 대체로 높은 항균 활성을 보였다. 특히 시제품 BP1과 B2의 활성이 높았다. Referring to FIG. 1 , whether the mixture sample was a mixture of ethanol extract or a mixture of essential oil extracts, it could be seen that each sample had higher antibacterial activity than a single sample, and the prototype sample showed generally higher antibacterial activity as the amount of sample added increased. In particular, the activity of the prototypes BP1 and B2 was high.
기타 유해 미생물에 대한 항균 활성 결과를 도 3 내지 도 5에 나타내었다. 도 3은 에탄올 추출물의 항균 활성 결과이고, 도 4는 정유 추출물의 항균 활성 결과이며, 도 5는 시제품의 항균 활성 결과이다.The results of antibacterial activity against other harmful microorganisms are shown in FIGS. 3 to 5 . Figure 3 is the result of the antibacterial activity of the ethanol extract, Figure 4 is the result of the antibacterial activity of the essential oil extract, Figure 5 is the result of the antibacterial activity of the prototype.
대부분의 시료가 정도의 차이는 있지만 대체로 항균 활성을 나타내었으며, 특히 정유 추출물과 시제품 BP1과 BP2가 활성이 높았다.Most of the samples showed antibacterial activity to some degree, but especially the essential oil extracts and prototypes BP1 and BP2 showed high activity.
2.2 시제품 세균수 측정 결과2.2 Prototype bacterial count results
시제품 가글액에 대한 세균수 측정 결과를 도 6에 나타내었다. 여기서 시료는 가장 항균 활성이 높은 시제품 BP1만을 사용하였다. 세균수 측정 결과 가글 전후 구강내 세균수는 LB 배지와 BHI 배지 모두에서 뚜렷하게 감소하였음을 알 수 있다. Fig. 6 shows the result of measuring the number of bacteria for the prototype gargle solution. Here, only the prototype BP1 with the highest antibacterial activity was used as the sample. As a result of measuring the number of bacteria, it can be seen that the number of bacteria in the oral cavity before and after gargling was significantly decreased in both the LB medium and the BHI medium.
2.3 구강내 구취가스 분석 실험 결과2.3 Oral bad breath gas analysis test results
상기 혼합물 시료에 대해서 구강내 구취가스 분석 실험 결과를 도 7에 나타내었다. 여기서도 시료는 가장 항균 활성이 높은 시제품 BP1만을 사용하였다. BP1은 양성대조군인 Listerine과 유사하게 구취 제거 효과가 있었다. 7 shows the results of an oral bad breath gas analysis experiment for the mixture sample. Here, only the prototype BP1 with the highest antibacterial activity was used as the sample. BP1 had the effect of removing bad breath similar to the positive control, Listerine.
2.4 관능평가 결과2.4 Sensory evaluation results
관능평가 결과를 아래의 표에 나타내었다. 관능평가도 BP1만을 가지고 수행하였다. The sensory evaluation results are shown in the table below. Sensory evaluation was also performed with only BP1.
시중에 판매되는 리스테린(녹차첨가)의 경우 자극성이 강하다는 의견이 많았으나 개발된 시제품 BP1는 자극성이 적고 전체적인 기호도가 양호한 것으로 조사되었다. 시제품 CM4는 린스테린보다 우수한 제품임을 확인하였다.In the case of commercially available Listerine (with green tea added), there were many opinions that it was highly irritating, but the developed prototype BP1 had less irritation and was found to have good overall acceptance. It was confirmed that the prototype CM4 was superior to linsterin.
Claims (5)
상기 추출물은 물과 에탄올의 혼합용매 추출물 또는 정유 추출물인 것을 특징으로 하는 스트렙토코커스 뮤탄스 균에 대한 항균 조성물.
A mixture of arborvitae leaf extract and loquat leaf extract as an active ingredient,
The extract is an antibacterial composition against Streptococcus mutans, characterized in that the extract is a mixed solvent extract of water and ethanol or an essential oil extract.
상기 상기 조성물은 구강청결제 조성물 또는 치약 조성물인 것을 특징으로 하는 조성물.
According to claim 1,
The composition is a mouthwash composition or a toothpaste composition, characterized in that the composition.
상기 추출물은 물과 에탄올의 혼합용매 추출물 또는 정유 추출물인 것을 특징으로 하는, 스트렙토코커스 뮤탄스 균에 의한 치아우식증 개선용 조성물.
A mixture of arborvitae leaf extract and loquat leaf extract as an active ingredient,
The extract is a composition for improving dental caries caused by Streptococcus mutans, characterized in that the extract is a mixed solvent extract of water and ethanol or an essential oil extract.
상기 상기 조성물은 구강청결제 조성물 또는 치약 조성물인 것을 특징으로 하는 조성물.
4. The method of claim 3,
The composition is a mouthwash composition or a toothpaste composition, characterized in that the composition.
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