KR20020082308A - Composition for the mouth containing extraction of Euonymus alatus Sieb - Google Patents
Composition for the mouth containing extraction of Euonymus alatus Sieb Download PDFInfo
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- KR20020082308A KR20020082308A KR1020010021516A KR20010021516A KR20020082308A KR 20020082308 A KR20020082308 A KR 20020082308A KR 1020010021516 A KR1020010021516 A KR 1020010021516A KR 20010021516 A KR20010021516 A KR 20010021516A KR 20020082308 A KR20020082308 A KR 20020082308A
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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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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- 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
Abstract
Description
본 발명은 귀전우 추출물을 함유하는 구강용 조성물에 관한 것으로 더욱 상세하게는 귀전우 추출물을 함유시킴으로서 구강내 미생물의 생육을 억제시켜 풍치 및 치주질환을 효과적으로 예방 및 치료하기 위한 귀전우 추출물을 함유한 구강용 조성물에 관한 것이다.The present invention relates to a composition for oral cavity containing an extract of a cow's ear, more specifically, containing an ear cow's extract to inhibit the growth of microorganisms in the oral cavity containing an extract of the cow's ear to effectively prevent and treat taste and periodontal disease. It relates to a composition for oral cavity.
일반적으로, 치과 영역의 질환 중 치주질환은 발병요인도 복잡하고 증상도 다양하다. 이중 치주질환 및 잇몸 염증의 주요 발생원인의 하나가 구강내 미생물에 의한 것으로 이들은 일차적으로 당류등을 이용하여 불용성 글루칸을 생성하고 이들이 각종 균체와 같이 치아표면에 부착하여 치석을 형성하게 된다. 이러한 치석에 의해 충치 및 치주질환이 발생하게 된다.In general, periodontal disease among the diseases in the dental field is complicated and various symptoms. One of the major causes of periodontal disease and inflammation of the gums is caused by microorganisms in the oral cavity. They primarily produce insoluble glucan using sugars and the like and attach to the tooth surface like various cells to form tartar. This tartar causes tooth decay and periodontal disease.
따라서, 치주질환 및 잇몸 염증을 예방하기 위한 가장 효과적인 방법은 이러한 미생물들의 활성을 억제하고, 염증이 발생한 잇몸의 혈류를 좋게하는 물질을 사용하므로서 이루어 질 수 있다.Therefore, the most effective method for preventing periodontal disease and gum inflammation can be achieved by using a substance that inhibits the activity of these microorganisms and improves the blood flow of the inflamed gum.
즉, 치주질환의 예방에 대해서는 구강내 청결을 적절히 행하는 것이 중요하지만, 예방치료를 목적으로 여러 가지 약제를 배합시킨 구강용 조성물이 제안되고 있다.That is, it is important to properly clean the oral cavity for the prevention of periodontal disease, but a composition for oral cavity in which various drugs are formulated for the purpose of prophylactic treatment has been proposed.
특히, 치주질환의 예방치료를 목적으로 사용되고 있는 약제중 충치의 주요 원인균인 스트렙토코커스 뮤탄스의 생육을 억제하고 살균하는데 이용되는 페니실린, 에리스로마이신, 테트라사이클린과 같은 항생제는 살균 및 생육 억제에는 효과적이기는 하나 내성균의 출현을 유발하는 등의 부작용이 크다.In particular, antibiotics such as penicillin, erythromycin, and tetracycline, which are used to inhibit and sterilize the growth of Streptococcus mutans, a major causative agent of tooth decay, are used for the prevention and treatment of periodontal disease. However, side effects such as causing the emergence of resistant bacteria are great.
또한, 클로르헥시딘과 같은 항균제는 충치 형성을 저해하는 효과는 있으나, 독성이 강하고 치아착색등과 같은 부작용이 있는 것으로 알려졌다. (J.Dect.Res.23,441(1946), J.Dent.Res.31,421(1966), J.Oral Ther. Pharmacol. 3,157(1966), J.Am.Dent.Ass.871006(1973))In addition, an antimicrobial agent such as chlorhexidine has an effect of inhibiting the formation of caries, but it is known to have high toxicity and side effects such as tooth coloring. (J. Dect. Res. 23,441 (1946), J. Dent. Res. 31,421 (1966), J. Oral Ther. Pharmacol. 3,157 (1966), J. Am. Dent. Ass. 871006 (1973))
따라서, 상기의 문제점을 해결하기 위해 종래의 치마분 등의 구강용 조성물 성분에 약효성분, 방부제, 안정화제 성분이 첨가된 구강용 조성물이 개발되고 있고, 구강내의 세정작용에 더해 약리작용 및 안정성이 우수한 구강용 조성물 등이 제공되고 있다.Therefore, in order to solve the above problems, a composition for oral cavity in which a medicinal ingredient, an antiseptic, and a stabilizer component is added to a composition for oral cavity, such as a dentifrice, has been developed. Oral compositions and the like are provided.
이중 대표적인 약효성분의 예가 천연 유래의 물질들로 생귀나리아(Sanguinaria)추출물(미합중국 특허 4,689,216 호), 몰약(Myrrh), 라타니(Rhatany), 세이지(Sage), 카모밀레(Chamomile), 에치네시아(Echinacea)등의 추출물(S. de Rysky ; Journal of Clinical Dentistry Vol.I, Supplement A. A22-24, 1988), 세이지(Sage) 및 미질향(Rosemary)등의 추출물(일본국 공개 특허 공보 소58-134,013호), 홉(Hops) 추출물(일본국 공개 특허 공보 소 62-138,420 호)등이 알려져 있다.Examples of representative active ingredients include Sanguinaria extract (US Pat. No. 4,689,216), Myrrh, Rhatany, Sage, Chamomile, Echine Extracts such as Echinacea (S. de Rysky; Journal of Clinical Dentistry Vol.I, Supplement A. A22-24, 1988), Extracts such as Sage and Rosemary (Japanese Patent Laid-Open Publication) 58-134,013), Hops extract (Japanese Patent Laid-Open No. 62-138,420) and the like are known.
그러나, 이들은 그들의 불안정성으로 인해 제제 혹은 구강중에서 실활되어 충분한 효과를 나타내지 못하는 문제점이 있었다.However, they have a problem in that they are inactivated in the preparation or the oral cavity due to their instability and do not exhibit sufficient effects.
따라서, 본 발명의 목적은 상기와 같은 문제점을 해결하기 위한 것으로, 귀전우 추출물이 구강용 조성물에 배합됨으로써 종래 구강 조성물의 불안정성으로 인해 제제 혹은 구강중에서 실활되는 문제점을 극복하기 위한 효과적인 구강 조성물을 제공하는 것이다.Accordingly, an object of the present invention is to solve the problems as described above, by providing the oral cavity extract to the composition for oral cavity to provide an effective oral composition for overcoming the problem of inactivation in the formulation or oral cavity due to the instability of the conventional oral composition. It is.
본 발명의 다른 목적 및 적용은 하기 발명의 상세한 설명으로부터 당업자에게 명백하게 드러날 것이다.Other objects and applications of the present invention will become apparent to those skilled in the art from the following detailed description.
본 발명은 귀전우 추출물을 함유하는 구강용 조성물에 관한 것이다.The present invention relates to a composition for oral cavity containing the extract of precious cow.
본 발명에 따른 귀전우에서 물, 메탄올, 아세톤, 1.3-부틸렌글리콜 또는 이들의 혼합물로 추출되어 얻어진 귀전우 추출물을 총 중량에 대해 0.01-5.0Wt% 함유함을 특징으로 한다.It is characterized in that it contains 0.01-5.0 Wt% of the total weight of the ear cow extract obtained by extraction with water, methanol, acetone, 1.3-butylene glycol or a mixture thereof in the ear cow according to the present invention.
본 발명에 사용되는 귀전우 추출물은 충치의 주요 원인균에 대해 항균 효과를 갖는 성분으로, 예로부터 한방이나 민간에서 산후 피멎이약, 정신불안, 여성의자궁출혈, 대하, 어혈을 없애는 약으로 쓰였으며, 열매를 오래 달여 고약을 만들어 피부병 치료에 사용해 왔다. 그외 혈액순환을 좋게 하고 어혈을 풀어 주며 염증을 없애고 정신을 안정시켜 주는 등의 약효를 갖고 있다고 알려져 있다.The extract from the ear cow used in the present invention has an antimicrobial effect on the main causative agent of tooth decay, and has been used as a medicine to remove postpartum blood pill, mental anxiety, uterine bleeding, lobulation, and blood from the Chinese medicine. The fruit has been decocted for a long time and used to treat skin diseases. In addition, it is known that it has good effects such as improving blood circulation, releasing blood, removing inflammation and stabilizing the mind.
특히, 본초학에서는 귀전우의 약효로 혈당강하작용, 혈전억제, 지혈, 치풍, 통월경 및 광증을 들고 있다.In particular, the herbal medicine has a hypoglycemic effect, thrombosis suppression, hemostasis, typhoon, dysmenorrhea and madness.
이러한 귀전우는 노박덩굴과(Celastraceae)에 속하는 화살나무의 날개가 붙어있는 간지를 채취한 것으로 우리나라 전역에 식재 또는 자생분포하며 일본, 중국에도 분포한다.These deciduous cattle are collected from the leaves of arrowheads belonging to the celticaceae family and are planted or distributed natively throughout Korea and distributed in Japan and China.
귀전우의 주요 성분은 만니톨(Mannitol), 솔비톨(Sorbitol), 글리세롤(Glycerol), 당(Sugar), 키리산테민(chrysanthemin), 벤조익액시드(benzoic acid), 둘시톨(Dulcitol), 탄닌(Tannin), 고무질 등이며 애피프리델리놀, 프리델라놀등의 테르페노이드도 함유하고 있다.The main components of guinea pigs are mannitol, sorbitol, glycerol, sugar, sugar, chrysanthemin, benzoic acid, dulcitol, and tannin. ), Rubber, etc., and also contains terpenoids such as epipridelinol and pridelanol.
상기 테르페노이드는 불포화 탄화수소의 일종으로 항균, 살충작용등이 보고되고 있으며 피부자극제, 소염제, 소독제, 완화제로 이용되고 있다.The terpenoids are a kind of unsaturated hydrocarbons, which have been reported to have antibacterial and insecticidal effects and are used as skin irritants, anti-inflammatory agents, disinfectants, and emollients.
또한, 항생, 항암, 혈압강화, 호르몬 분비촉진 작용을 가지며 뇌를 자극해 흥분 또는 진정 작용을 가지므로 향기요법에도 쓰이는 물질이다.In addition, antibiotics, anti-cancer, blood pressure strengthening, hormonal secretion action and stimulate the brain to have an excitement or sedative effect, so it is used in aromatherapy.
따라서, 이상에서 언급된 귀전우 추출물의 구강질환에 대한 유효성을 검토한 결과, 본 발명자는 충치원인균인 스트렙토코커스 뮤탄스(Streptococcus mutans ATCC 27607, ATCC 27341, ATCC 25175,NRRL 10449), 스트렙토코커스 생기스(Streptococcus sanguis), 스트렙토코커스 살리베리어스(Streptococcussalivarius) 및 치주병 원인균인 악티노마이세스 비스코커스(Actinomyces viscous), 악티노마이세스 내스런디( Actinomyces naeslundi), 박테리오데스 깅기발라스(Bacteriodes gingivalis), 박테리오데스 프로시디어스(Bacteriodes frosythus), 박테리오데스 인터메디어스(Bacteriodes intermedius)에 대하여 항균작용이 있음을 확인하였다.Therefore, as a result of examining the effectiveness of oral disease of the above-mentioned ear cow extract, the present inventors have identified Streptococcus mutans ATCC 27607, ATCC 27341, ATCC 25175, NRRL 10449 (Streptococcus sanguis), Streptococcus salivarius, and Actinomyces viscous, Actinomyces naeslundi, and Bacteriodes gingivalisivaivagingus , Bacteriodes frosythus, Bacteriodes intermedius (Bacteriodes intermedius) was confirmed to have an antimicrobial action.
본 발명에 따른 구강용 조성물은 귀전우 추출물을 함유함으로써 프라그 형성이 억제되고, 충치 및 치주질환이 예방된다.The composition for oral cavity according to the present invention contains plaque extract, inhibits plaque formation, and prevents tooth decay and periodontal disease.
이하 본 발명을 상세히 설명하면 다음과 같다.Hereinafter, the present invention will be described in detail.
본 발명은 구강용 조성물 중에 귀전우 추출물이 배합되었을때의 구강 조성물에 관한 것이다.The present invention relates to an oral composition when the precious cow extract is blended in the composition for oral cavity.
상기 귀전우추출물의 추출시 물, 에탄올, 메탄올, 아세톤, 1.3-부틸렌글리콜등의 용매를 그대로 또는 혼합하여 귀전우 원료에 대해 2-10배를 넣어 추출한 후, 여과액을 농축하여 농축물을 얻었다. 이렇게 하여 얻은 귀전우 추출액의 구강용 조성물의 배합량은 전체의 0.01-5.0Wt％, 바람직하게는 0.5-3.0Wt％로 배합하는 것이 좋다.When extracting the precious cow extract, a solvent such as water, ethanol, methanol, acetone, 1.3-butylene glycol or the like is mixed and extracted 2-10 times with respect to the raw cow cattle, and the filtrate is concentrated to concentrate the concentrate. Got it. Thus, the compounding quantity of the composition for oral cavity of the obtained cow's milk extract is 0.01-5.0 Wt% of the whole, Preferably it is good to mix | blend 0.5-3.0Wt%.
귀전우 추출물의 배합량이 0.01Wt％미만이면 치주질환 예방, 치료 및 충치예방효과가 미약하게 되고, 5.0Wt％ 이상일 경우에는 큰 효과를 기대할 수 없다.If the amount of the extract of less than 0.01 Wt%, the effect of preventing, treating, and preventing tooth decay of the periodontal disease is weak.
본 발명에 따른 귀전우 추출물은 수분을 함유하는 각종의 조성물, 예를 들면, 크림, 치마제 등에 배합하는 것이 가능하다.The deciduous cow extract according to the present invention can be blended with various compositions containing moisture, for example, creams, dentifrices and the like.
따라서, 이하 구강용 조성물의 필수성분과 첨가물에 대하여 설명한다. 이들성분의 양은 당업자에 의해 용이하게 선정될 수 있다.Therefore, the essential components and additives of the composition for oral cavity are described below. The amount of these components can be easily selected by those skilled in the art.
본 발명에서 실시하기 위한 각종 구강용 조성물은 그 종류에 따라 적당한 성분을 사용해야 하는데, 예를 들면 통상적인 치약성분들로서 연마제, 습윤제, 기포제, 결합제, 감미제, pH조절제, 방부제, 약효성분, 향료, 증백제, 색소, 용제등을 함유할 수 있다.Various oral compositions for carrying out the present invention should use a suitable component according to the type, for example, as a conventional toothpaste components, abrasives, wetting agents, foaming agents, binders, sweeteners, pH adjusting agents, preservatives, medicinal ingredients, fragrances, thickening A white agent, a pigment, a solvent, etc. can be contained.
상기 연마제로는 탄산칼슘, 침강 실리카, 수산화알루미늄, 인산일수소칼슘, 불용성 메타인산나트륨 등이 있으며, 이들 연마제를 단독 혹은 2종 이상 혼합하여 1-60Wt％, 바람직하게는 10-50Wt％를 사용한다.Examples of the abrasive include calcium carbonate, precipitated silica, aluminum hydroxide, calcium monohydrogen phosphate, and insoluble sodium metaphosphate. These abrasives may be used alone or in combination of two or more of them, using 1-60 Wt%, preferably 10-50 Wt%. do.
상기 습윤제로서는 글리세린, 소르비톨액, 폴리에틸렌글리콜, 프로필렌글리콜 등을 단독 혹은 2종 이상 혼합하여 10-60Wt％, 바람직하게는 20-50Wt％를 사용한다.As the humectant, 10-60 Wt%, preferably 20-50 Wt%, of glycerin, sorbitol solution, polyethylene glycol, propylene glycol, or the like may be used alone or in combination of two or more thereof.
상기 기포제로는 라우릴황산나트륨, 소디움라우릴사르코시네이트, 자당지방산에스테르, 폴리옥시에칠렌경화피마자유, 폴리옥시에칠렌폴리옥시프로필렌 공중합체 등의 음이온 및 비이온 계면활성제를 단독 혹은 2종 이상 혼합하여 0.5-5.0Wt％, 바람직하게는 0.5-3.0Wt％를 사용한다.As the foaming agent, anionic and nonionic surfactants such as sodium lauryl sulfate, sodium lauryl sarcosinate, sucrose fatty acid ester, polyoxyethylene-cured castor oil, and polyoxyethylene polyoxypropylene copolymer may be used alone or in combination of two or more thereof. 0.5-5.0 Wt%, preferably 0.5-3.0 Wt% is used.
상기 결합제로서는 카르복시메칠 셀룰로오스 나트륨, 하이드록시메틸셀룰로오스, 카라기난, 잔탄껌, 알긴산 나트륨등을 단독 혹은 2종 이상 혼합하여 0.1-5.0Wt％, 바람직하게는 0.1-2.0Wt％를 사용한다.As said binder, 0.1-5.0 Wt%, Preferably 0.1-2.0 Wt% is used individually or in mixture of 2 or more types of carboxymethyl cellulose sodium, hydroxymethylcellulose, carrageenan, xanthan gum, and sodium alginate.
상기 감미제로는 삭카린나트륨, 아스파탐, 스테비오사이드, 자이리톨, 감초산등을 단독 혹은 2종 이상 혼합하여 0.05-5.0Wt％를 사용하는 것이 바람직하다.As the sweetener, it is preferable to use 0.05-5.0 Wt% of saccharin sodium, aspartame, stevioside, ziitol, and licorice acid alone or in mixture of two or more thereof.
상기 pH조절제로는 인산나트륨, 인산이나트륨, 인산삼나트륨, 피로인산나트륨, 구연산, 구연산나트륨, 주석산등이 있다The pH adjusting agent includes sodium phosphate, disodium phosphate, trisodium phosphate, sodium pyrophosphate, citric acid, sodium citrate, tartaric acid, and the like.
상기 방부제로는 파라옥시안식향산메칠, 파라옥시안식향산프로필, 안식향산나트륨을 단독 혹은 2종 이상 혼합하여 사용한다.As said preservative, paraoxybenzoic acid methyl, propyl paraoxybenzoic acid, and sodium benzoate are used individually or in mixture of 2 or more types.
상기 약효성분으로는 불화나트륨, 일불소인산나트륨, 불화주석, 클로로헥시딘, 알란토인클로로히드록시알루미네이트, 아미노카프론산, 트라넥사민산, 트리클로산, 염화세틸피리디움, 염화아연, 염산피리독신, 초산토코페롤등을 단독 혹은 2종 이상 혼합하여 사용한다.Examples of the active ingredient include sodium fluoride, sodium monofluorophosphate, tin fluoride, chlorohexidine, allantoin chlorohydroxy aluminate, aminocaproic acid, tranexamic acid, triclosan, cetylpyrididium, zinc chloride, pyridoxine hydrochloride, Tocopherol acetate, etc. may be used alone or in combination of two or more thereof.
상기 향료로는 페퍼민트오일, 스피아민트오일, 멘톨, 아네톨등을 적당량 혼합사용한다.As the fragrance, peppermint oil, spearmint oil, menthol, anetol and the like are mixed and used in an appropriate amount.
상기 색소로는 식용색소를, 정제수 및 에탄올의 용제로 통상의 구강용 조성물의 제조방법에 따라 제조할 수 있다.As said pigment, food colorings can be manufactured according to the manufacturing method of the composition for oral cavity in the solvent of purified water and ethanol.
본 발명에서 귀전우 추출물 제조방법은 다음과 같다.In the present invention, the method of producing a jeonwoowoo extract is as follows.
본 발명에서 귀전우 추출물의 제조방법은 화살나무의 잎과 여린 가지(Twigs) 1킬로그램을 1.0 내지 2.0 센티미터로 절단한 다음, 이를 용매 속에 24시간 이상 침적시키고 이를 압착 여과하여 얻는다.In the present invention, the method for producing a jeonwoowoo extract is obtained by cutting one kilogram of leaves and tender branches (Twigs) to 1.0 to 2.0 centimeters, immersed in a solvent for 24 hours or more and compression filtration.
이때, 사용한 용매는 1,3-부틸렌글리콜과 에탄올(95%)의 중량비가 1:1 내지 1:2가 되게 하고 화살나무의 잎과 여린 가지와 용매의 중량비가 1:1 내지 1:3으로 한다.In this case, the solvent used is a weight ratio of 1,3-butylene glycol and ethanol (95%) is 1: 1 to 1: 2, and the weight ratio of the leaves and fragile branches of the arrow and the solvent is 1: 1 to 1: 3. It is done.
이하 본 발명의 구강용 조성물의 효과에 대하여 시험 예 및 실시 예를 들어상세히 설명할 것이나, 이들 실시 예에 의해 본 발명의 범위가 한정되는 것은 아니다.Although the test example and the Example will be described in detail about the effect of the composition for oral cavity of this invention below, the scope of the present invention is not limited by these Examples.
[시험예 1][Test Example 1]
구강용 조성물에 사용한 귀전우 추출물의 충치 및 치주질환균에 대한 항균효과를 하기 실험방법에 따라 측정하고, 그 결과를 표 1에 나타낸다.The antimicrobial effect of the decayed ear extract used in the composition for oral cavity against caries and periodontal disease bacteria was measured according to the following experimental method, and the results are shown in Table 1.
1) 실험방법1) Experiment Method
시험균인 박테리오데스 깅기발라스(Bacteriodes gingivalis ATCC 33277)와 후소박테리움 뉴클레아툼(Fusobacteriumnucleatum ATCC 25586), 스트렙토코커스 뮤탄스(Streptococcus mutans ATCC 27607), 악티노마이세스 비스코커스(Actinomyces viscosus ATCC 15987)를 BHI(Brain Heart Infusion) 배지에 각각 배양하고, BHI배지에 시료인 귀전우추출물을 농도별로 연속 희석하여 최종농도가 5 ppm이 되도록 하였다.Test strains Bacteriodes gingivalis ATCC 33277, Fusobacterium nucleatum ATCC 25586, Streptococcus mutans ATCC 27607, Actinomyces Viscous 1569 ) Was incubated in BHI (Brain Heart Infusion) medium, and the sample of the degenerative cow extract, which was sampled in BHI medium, was continuously diluted to 5 ppm.
상기에서 제조된 시험균 현탁액을 접종하여 각각의 희석된튜브에 세균수가 103cells/정도 되도록 한 후, 37도에서 24시간 배양하여 세포의 성장을 저해하는 최소균성장 억제농도(MIC : minimal inhibitory concentration)를 측정하였다.After inoculating the test cell suspension prepared above, the number of bacteria in each diluted tube was about 103 cells /, followed by culturing at 37 ° C. for 24 hours to determine the minimum inhibitory concentration (MIC) that inhibits the growth of cells. Measured.
이때, 후소박테리움 뉴클레아툼(Fusobacterium nucleatum ATCC 25586), 악티노마이세스 비스코커스(Actinomycesviscosus ATCC 15987)는 혐기성자(Anaerobic chamber)에서 배양하였다.At this time, Fusobacterium nucleatum ATCC 25586 and Actinomyces vicosus ATCC 15987 were incubated in an anaerobic chamber.
[표 1] 실시예와 비교예의 최소저해농도(MIC)TABLE 1 Minimum Inhibitory Concentration (MIC) of Examples and Comparative Examples
[시험예 2][Test Example 2]
일반적으로, 충치는 구강내 미생물인 스트렙토코커스 뮤탄스에 의해 발생하는 것으로 알려져 있다.Generally, caries is known to be caused by Streptococcus mutans, an oral microorganism.
즉, 이 균의 균체외 효소인 글루코실 트랜스퍼레이즈(GTase)는 음식을 섭취한 후 구강내 남아 있는 각종 당류로부터 접착성이 있는 다당체의 일종인 수용성 및 불용성 글루칸을 합성하는데, 이들 중 특히 불용성 글루칸이 각종 세균들과 더불어 치아표면에 부착하여 견고한 치석을 형성하게 된다. 이들 치석 내에 부착된 각종 세균들은 구강 내에 잔조해 있는 각종 유기물들을 대사하고, 그 결과로 생성된 유기산에 의해 치아 표면을 둘러싸고 있는 에나멜을 용해함으로써 치아부식이 일어나게 된다.In other words, the bacterium's extracellular enzyme glucosyl transferase (GTase) synthesizes water-soluble and insoluble glucan, which is a kind of adhesive polysaccharide, from various sugars remaining in the oral cavity after ingesting food. Together with these bacteria, they attach to the tooth surface to form solid tartar. Various bacteria attached to these tartars metabolize various organic matter remaining in the oral cavity, and tooth corrosion occurs by dissolving enamel surrounding the tooth surface by the resulting organic acid.
따라서, 본 실험에서는 충치균이 내는 균체의 효소인 글루코실 트랜스퍼레이즈(GTase)가 불용성 글루칸을 합성하는 과정을 억제함으로써, 충치 예방목적을 달성할 수 있다고 생각하여, 귀전우 추출물이 함유된 구강요조성물의 (실시예 1-4) 글루코실 트랜스퍼레이즈(GTase) 저해 효과를 측정하였다.Therefore, in this experiment, it is thought that glucosyl transferase (GTase), which is an enzyme of the cells of caries, inhibits the process of synthesizing insoluble glucan, thereby achieving the purpose of preventing tooth decay. (Example 1-4) Glucosyl transferase (GTase) inhibitory effect of was measured.
1) 실험방법1) Experiment Method
치약 베이스에서의 글루코실 트랜스퍼레이즈(GTase) 저해효과 측정.Determination of Glucosyl Transferase (GTase) Inhibitory Effect in Toothpaste Base.
① 글루코실 트랜스퍼레이즈(GTase) 분리 및 정제① Glucosyl Transferase (GTase) Isolation and Purification
스트렙토코커스 뮤탄스 ATCC 25175를 3ℓ의 BHI 브로쓰에 접종하여 37℃ 인큐베이터에서 18시간 배양한 후 원심분리(3000rpm, 20분)하여 균체를 제거하고, 그 상층액을 황산암모늄으로 60-70％포화시킨 후 4℃에서 18시간동안 방치하였다. 이렇게 하여 생성된 침전물을 원심분리(3000rpm, 20분)하여 인산칼륨완충액(0.05M pH6.5) 200ml에 용해한 후 동일 완충용액으로 24시간 투석시켰다. 투석이 완료된 액을 다시 원심분리하여 불용물을 제거하고, 얻은 조효소 용액을 히드록시아파타이트(hydroxyapatite)로 충진된 칼럼에 주입한 후 이온강도가 다른 동일 완충용액을 단계적으로 용출시켜 정제된 글루코실 트랜스퍼레이즈(GTase)를 얻었다.Streptococcus mutans ATCC 25175 was inoculated in 3 L BHI broth, incubated for 18 hours in a 37 ° C incubator, centrifuged (3000 rpm, 20 minutes) to remove the cells, and the supernatant was saturated with ammonium sulfate 60-70%. After standing for 18 hours at 4 ℃. The precipitate thus produced was centrifuged (3000 rpm, 20 minutes), dissolved in 200 ml of potassium phosphate buffer (0.05 M pH6.5), and dialyzed with the same buffer solution for 24 hours. The dialysis solution was centrifuged again to remove insoluble materials, and the obtained coenzyme solution was injected into a column filled with hydroxyapatite, followed by stepwise eluting of the same buffer solution having different ionic strength, thereby purifying glucosyl transfer. GTase was obtained.
② 글루코실 트랜스퍼레이즈(GTase) 저해활성 측정② Measurement of Glucosyl Transferase (GTase) Inhibitory Activity
상기방법으로 제조된 GTF 50-100㎍, 설당 70㎍및 0.02％ NaN3를 함유하는 0.05M인산칼륨완충액(pH 6.5) 4ml에 실시예 1-4의 구강용 조성물 각각 10g을 증류수로 농도 50％까지 희석한 다음, 원심분리하여 상등액을 얻는다.4 g of 0.05 M potassium phosphate buffer solution (pH 6.5) containing 50 to 100 μg of GTF prepared by the above method, 70 μg of sugar and 0.02% NaN 3, each of 10 g of the oral composition of Example 1-4 was diluted to 50% in distilled water. Dilute and then centrifuge to obtain supernatant.
이 액을 농도 0.5％까지 재희석한 후, 완충용액에 가하고 37℃에서 18시간 동안 항온기에서 반응시킨 후 생성된 글루칸의 양을 분광광도계(spectrophotometer)로 660㎚에서 흡광도를 측정하였다.After diluting the solution to a concentration of 0.5%, it was added to a buffer solution and reacted at 37 ° C. for 18 hours in an incubator, and then the absorbance was measured at 660 nm using a spectrophotometer.
이 파장에서 흡광도의 감소는 글루칸의 생성 억제, 즉 글루코실 트랜스퍼레이즈(GTase) 저해도에 비례한다. 글루코실 트랜스퍼레이즈(GTase) 저해활성도는 다음 식에 의해 구하였으며, 대조용액의 흡광도는 귀전우 추출물을 제외한 반응액의 흡광도이다.The decrease in absorbance at this wavelength is proportional to the inhibition of glucan production, ie the inhibition of glucosyl transferase (GTase). Glucosyl transferase (GTase) inhibitory activity was determined by the following equation, and the absorbance of the control solution was the absorbance of the reaction solution except for the extract of cow ox.
글루코실 트랜스퍼레이즈(GTase) 저해활성도(％) = {1- ( X / Y ) x 100}Glucosyl Transferase (GTase) Inhibitory Activity (%) = {1- (X / Y) x 100}
( X : 시료용액의 흡광도 Y : 대조용액의 흡광도 )(X: absorbance of sample solution Y: absorbance of control solution)
2) 실험결과2) Experiment result
상기 방법에 의해 표 2 와 같은 결과를 얻었다. 즉 귀전우 추출물이 함유된 구강용조성물(실시예 1-3)은 글루코실 트랜스퍼레이즈(GTase) 저해 효과가 우수하므로, 충치 예방을 목적으로 하는 구강용 조성물에 효과적이다.The result shown in Table 2 was obtained by the said method. That is, the composition for oral cavity (Examples 1-3) containing an extract of an ear cow has excellent glucosyl transferase (GTase) inhibitory effect, and thus is effective for oral compositions for the purpose of preventing tooth decay.
[표 2] 조성물 중에서 귀전우 추출물에 의한 글루코실 트랜스퍼레이즈(GTase) 저해 효과.[Table 2] Glucosyl transferase (GTase) inhibitory effect by the extract from cows in the composition.
실시예 1 : 치약Example 1: Toothpaste
실시예 2 : 불투명 치약Example 2: Opaque Toothpaste
실시예 3 : 구강세정제Example 3: Mouthwash
실시예 4 : 구강청량제Example 4 Oral Refresher
본 발명에 따른 구강용 조성물은 귀전우 추출물을 함유하여 구강내 미생물의 생육을 억제시켜 충치 및 치주질환의 예방 및 치료에 효과적이며, 저농도에서도 충분한 효과를 발휘할 수 있다.The composition for oral cavity according to the present invention is effective in the prevention and treatment of tooth decay and periodontal disease by suppressing the growth of microorganisms in the oral cavity by containing the extract of the ear canal, it can exhibit a sufficient effect even at low concentrations.
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| KR101592549B1 (en) | 2015-06-29 | 2016-02-18 | 김용수 | Composition for treating and preventing periodontal disease and caries |
| KR20160061934A (en) | 2016-05-13 | 2016-06-01 | 주식회사 파미니티 | Composition for preventing or alleviating periodontal disease |
| KR101641215B1 (en) * | 2015-05-15 | 2016-07-20 | 대한민국(환경부 국립생물자원관장) | Oral composition comprising Morus alba L. extract and Winged spindle extract for preventing and treating dental caries |
| KR101641213B1 (en) * | 2015-07-08 | 2016-07-20 | 대한민국(환경부 국립생물자원관장) | Oral composition comprising Morus alba L. extract and Winged spindle extract for preventing and treating periodontal disease |
| KR102078414B1 (en) * | 2018-10-31 | 2020-02-17 | 주식회사 발효예스 | METHOD FOR MANUFACTURING Euonymus alatus FERMENTED BROTH, AND THE Euonymus alatus FERMENTED BROTH MANUFACTURED BY THE METHOD |
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- 2001-04-20 KR KR1020010021516A patent/KR20020082308A/en not_active Application Discontinuation
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| KR101641215B1 (en) * | 2015-05-15 | 2016-07-20 | 대한민국(환경부 국립생물자원관장) | Oral composition comprising Morus alba L. extract and Winged spindle extract for preventing and treating dental caries |
| WO2016186335A3 (en) * | 2015-05-15 | 2017-01-12 | 대한민국(환경부 국립생물자원관장) | Oral composition for cavity prevention and treatment including mulberry root extract and euonymus alatus extract as effective ingredients |
| CN107073061A (en) * | 2015-05-15 | 2017-08-18 | 大韩民国（环境部国立生物资源馆） | The prevention of dental caries and treatment oral cavity composition containing the mulberry bark extract as active component and winged euonymus extract |
| KR101592549B1 (en) | 2015-06-29 | 2016-02-18 | 김용수 | Composition for treating and preventing periodontal disease and caries |
| KR101641213B1 (en) * | 2015-07-08 | 2016-07-20 | 대한민국(환경부 국립생물자원관장) | Oral composition comprising Morus alba L. extract and Winged spindle extract for preventing and treating periodontal disease |
| WO2017007140A1 (en) * | 2015-07-08 | 2017-01-12 | 대한민국(환경부 국립생물자원관장) | Oral composition for preventing and treating periodontitis, comprising mulberry root bark extract and winged spindle extract as active ingredients |
| CN106535911A (en) * | 2015-07-08 | 2017-03-22 | 大韩民国（环境部国立生物资源馆长） | Oral composition for preventing and treating periodontitis, comprising mulberry root bark extract and winged spindle extract as active ingredients |
| KR20160061934A (en) | 2016-05-13 | 2016-06-01 | 주식회사 파미니티 | Composition for preventing or alleviating periodontal disease |
| KR102078414B1 (en) * | 2018-10-31 | 2020-02-17 | 주식회사 발효예스 | METHOD FOR MANUFACTURING Euonymus alatus FERMENTED BROTH, AND THE Euonymus alatus FERMENTED BROTH MANUFACTURED BY THE METHOD |
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