KR20190138030A - Composition for improve stress comprising lactic acid bacteria and theanine - Google Patents
Composition for improve stress comprising lactic acid bacteria and theanine Download PDFInfo
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- KR20190138030A KR20190138030A KR1020180064069A KR20180064069A KR20190138030A KR 20190138030 A KR20190138030 A KR 20190138030A KR 1020180064069 A KR1020180064069 A KR 1020180064069A KR 20180064069 A KR20180064069 A KR 20180064069A KR 20190138030 A KR20190138030 A KR 20190138030A
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- stress
- lactobacillus
- theanine
- bifidobacterium
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Abstract
Description
본 명세서에는 유산균류 및 테아닌을 포함하는 스트레스 개선용 조성물이 개시된다.Disclosed herein is a composition for improving stress comprising lactic acid bacteria and theanine.
스트레스의 어원은 'Strictus : 팽팽한 혹은 좁은' 또는 'Stringere : 팽팽하다' 이며 심리학에서 스트레스 는 스트레스 요인에 대항하기 위한 심신의 변화과정을 뜻하고 우리말로는 정신압박 이라고도 한다. The etymology of stress is 'Strictus: Tense or Narrow' or 'Stringere: Tense'. In psychology, stress refers to the process of change of mind and body to counter stress factors, and in our words it is also called mental pressure.
스트레스는 일상에 만연되어 있는 요소로 유발되며, 거의 모든 질환과 의학적 질병의 발생에 기여하는 것으로 알려져 있다.Stress is caused by the prevailing elements of daily life, and is known to contribute to the occurrence of almost all diseases and medical diseases.
구체적으로, 스트레스를 유발하는 원인과 그로 인한 인체 내 생리적 변화는 다양하지만, 공통적으로 뇌하수체-부신계를 자극하여 스트레스 호르몬이 분비되고, 이러한 스트레스가 지속되면 인체의 기능이 저하되어 신경 과민, 두통, 소화 불량 등의 증상을 유발하며, 이것이 위십이지장궤양, 고혈압, 과민성대장염, 협심증 등의 2차 질병으로 발전된다. Specifically, the causes of stress and the physiological changes in the human body are various, but the stress hormones are secreted by stimulating the pituitary-adrenal system in common, and if such stress persists, the body's function is reduced, such as nervousness, headache, It causes symptoms such as indigestion, which develops into secondary diseases such as gastroduodenal ulcer, hypertension, irritable colitis, and angina pectoris.
지금까지 스트레스를 치유하기 위하여 항우울제, 항불안제 등의 인공 화합물을 포함하는 의약품을 사용하는 약물요법이 이용되고 있지만, 이러한 약물요법은 장기간 사용시 약물에 대한 내성 및 부작용 위험이 있어 지속적 이용이 불가능하므로, 부작용이 없고, 지속적으로 복용할 수 있는 방안에 대한 니즈가 높다.Until now, drug therapies using medicines containing artificial compounds such as antidepressants and anti-anxiety agents have been used to cure stress.However, these drug therapies have a resistance to side effects and risk of side effects and therefore are not continuously available. There is no need, and there is a high need for a continual dose.
이에 다양한 식물 추출물 등을 활용하는 방법이 계속해서 연구되고 있지만, 아직까지 만족할만한 효과를 얻은 연구 결과는 없는 실정이다.The use of various plant extracts, etc. have been studied continuously, but the results have not yet been obtained a satisfactory effect.
일 측면에서, 본 발명의 목적은, 스트레스를 효과적으로 개선할 수 있는 조성물을 제공하는 것이다.In one aspect, it is an object of the present invention to provide a composition that can effectively improve stress.
일 측면에서, 본 발명의 목적은, 천연물을 사용하여 인체에 대한 독성이 없는 스트레스 개선용 조성물을 제공하는 것이다.In one aspect, an object of the present invention is to provide a composition for improving stress that is not toxic to the human body using natural products.
일 측면에서, 본 발명의 목적은, 유산균과 테아닌 각각이 갖는 효능을 극대화하는 것이다.In one aspect, an object of the present invention, to maximize the efficacy of each of the lactic acid bacteria and theanine.
일 측면에서, 본 발명의 목적은, 유산균과 테아닌의 조합으로부터 극대화된 스트레스 완화 효과를 얻는 것이다.In one aspect, an object of the present invention, to obtain a maximum stress relief effect from the combination of lactic acid bacteria and theanine.
일 측면에서, 본 발명의 목적은, 스트레스로 인해 유발된 신체적 증상 및 심리적 증상을 완화 또는 개선시키는 것이다.In one aspect, the object of the present invention is to alleviate or ameliorate the physical and psychological symptoms caused by stress.
상기와 같은 목적을 달성하기 위하여, 본 발명은, 일 측면에서, 유산균류; 및 테아닌, 이의 약학적으로 허용 가능한 염, 이의 프로드럭, 이의 수화물 또는 이의 용매화물을 포함하는, 스트레스 개선용 조성물을 제공한다.In order to achieve the above object, the present invention, in one aspect, lactic acid bacteria; And theanine, a pharmaceutically acceptable salt thereof, a prodrug thereof, a hydrate thereof, or a solvate thereof.
또한, 일 측면에서, 본 발명은, 유산균류 및 테아닌을 포함하는 스트레스 개선용 조성물; 및 지시서를 포함하며, 상기 지시서는, 대상에 유산균류는 108~1010CFU/ day로 투여하고, 테아닌은 5~500mg/kg/day로 투여하는 내용을 포함하는, 스트레스 개선용 키트를 제공한다.In addition, in one aspect, the present invention, a stress improving composition comprising lactic acid bacteria and theanine; And instructions, wherein the instructions, the lactic acid bacteria to the subject is administered 10 8 ~ 10 10 CFU / day, theanine provides a content for stress improvement kit, including the contents administered at 5 ~ 500mg / kg / day do.
본 발명의 일 측면인 조성물은, 비장세포 증식 촉진능과 코르티코스테론 분비 억제능이 우수하여, 스트레스로 인한 신체적 증상 및 심리적 증상의 개선 효과가 우수하다. 또한, 상기 조성물은, 천연물을 사용하므로, 인체 독성이 없어 화장료, 식품 또는 약학 조성물로 활용도가 높다.The composition, which is an aspect of the present invention, is excellent in promoting splenocyte growth and inhibiting corticosteroid secretion, and is excellent in improving physical symptoms and psychological symptoms due to stress. In addition, since the composition uses a natural product, there is no human toxicity, and thus the composition is highly utilized as a cosmetic, food or pharmaceutical composition.
도 1 내지 도 3은 스트레스 유도 마우스의 혈중 코르티코스테론 농도 변화를 검측한 결과에 대한 것으로, 도 1은 L-테아닌, 락토바실러스 헬베티쿠스, 및 비피도박테리움 롱검을 각각 투여한 경우, 도 2와 3은 L-테아닌과 락토바실러스 헬베티쿠스, 및 비피도박테리움 롱검을 병용한 경우의 결과를 나타낸다.
도 1 내지 도 3에서, 락토바실러스 헬베티쿠스는 R52로 표시하고, 비피도박테리움 롱검은 R175로 표시하였다.
도 4 및 도 5는 스트레스 유도 마우스의 혈중 사이토카인 변화를 검측한 결과에 대한 것으로, 도 4는 IFN-γ 농도 변화 측정 결과, 도 5는 IL-4 농도 변화 측정 결과를 나타낸다.
도 6은, L-테아닌과 유산균류(락토바실러스 헬베티쿠스, 및 비피도박테리움 롱검)을 각각 투여한 경우, 또는 병용 투여한 경우 십자형 미로 실험 결과를 보이는 것이다.1 to 3 are results of detecting the change in blood corticosterone concentration of stress-induced mice, Figure 1 is a case of administration of L-theanine, Lactobacillus helveticus, and Bifidobacterium longgum, respectively 2 and 3 show the results of using L-theanine, Lactobacillus helvetus, and Bifidobacterium long gum in combination.
1 to 3, Lactobacillus helvetticus is indicated as R52 and Bifidobacterium longgum is indicated as R175.
4 and 5 show the results of detecting changes in the cytokine in the blood of stress-induced mice, FIG. 4 shows the results of measuring IFN-γ concentration changes, and FIG. 5 shows the results of measuring IL-4 concentration changes.
Figure 6 shows the results of a cross-shaped labyrinth when L-theanine and lactic acid bacteria (Lactobacillus helvetticus and Bifidobacterium longgum) were administered or co-administered, respectively.
이하, 첨부한 도면들을 참조하여, 본 출원의 실시예들을 보다 상세하게 설명하고자 한다. 그러나 본 출원에 개시된 기술은 여기서 설명되는 실시예들에 한정되지 않고 다른 형태로 구체화될 수도 있다. 단지, 여기서 소개되는 실시예들은 개시된 내용이 철저하고 완전해질 수 있도록 그리고 당업자에게 본 출원의 사상이 충분히 전달될 수 있도록 하기 위해 제공되는 것이다. 또한, 해당 분야에서 통상의 지식을 가진 자라면 본 출원의 기술적 사상을 벗어나지 않는 범위 내에서 본 출원의 사상을 다양한 다른 형태로 구현할 수 있을 것이다.Hereinafter, with reference to the accompanying drawings, it will be described embodiments of the present application in more detail. However, the technology disclosed in the present application is not limited to the embodiments described herein and may be embodied in other forms. It is merely to be understood that the embodiments introduced herein are provided so that the disclosure can be made thorough and complete, and that the spirit of the present application can be fully conveyed to those skilled in the art. In addition, one of ordinary skill in the art may implement the spirit of the present application in various other forms without departing from the technical spirit of the present application.
본 명세서에서 "약학적으로 허용 가능"이란 통상의 의약적 복용량(medicinal dosage)으로 이용할 때 상당한 독성 효과를 피함으로써, 동물, 더 구체적으로는 인간에게 사용할 수 있다는 정부 또는 이에 준하는 규제 기구의 승인을 받을 수 있거나 승인 받거나, 또는 약전에 열거되거나 기타 일반적인 약전으로 인지되는 것을 의미한다.본 명세서에서 "약학적으로 허용 가능한 염"은 약학적으로 허용 가능하고 모 화합물(parent compound)의 바람직한 약리 활성을 갖는 본 발명의 일측면에 따른 염을 의미한다. 상기 염은 (1) 염산, 브롬화수소산, 황산, 질산, 인산 등과 같은 무기산으로 형성되거나; 또는 아세트산, 프로파이온산, 헥사노산, 사이클로펜테인프로피온산, 글라이콜산, 피루브산, 락트산, 말론산, 숙신산, 말산, 말레산, 푸마르산, 타르타르산, 시트르산, 벤조산, 3-(4-하이드록시벤조일) 벤조산, 신남산, 만델산, 메테인설폰산, 에테인설폰산, 1,2-에테인-디설폰산, 2-하이드록시에테인설폰산, 벤젠설폰산, 4-클로로벤젠설폰산, 2-나프탈렌설폰산, 4-톨루엔설폰산, 캄퍼설폰산, 4-메틸바이사이클로 [2,2,2]-oct-2-엔-1-카르복실산, 글루코헵톤산, 3-페닐프로파이온산, 트리메틸아세트산, tert-부틸아세트산, 라우릴 황산, 글루콘산, 글루탐산, 하이드록시나프토산, 살리실산, 스테아르산, 뮤콘산과 같은 유기산으로 형성되는 산 부가염(acid addition salt); 또는 (2) 모 화합물에 존재하는 산성 프로톤이 치환될 때 형성되는 염을 포함할 수 있다.As used herein, "pharmaceutically acceptable" means the approval of a government or equivalent regulatory body to use in animals, more specifically in humans, by avoiding significant toxic effects when used in conventional medicinal dosages. Or pharmacologically acceptable salts herein means pharmaceutically acceptable salts that are pharmacologically acceptable and that provide for the desired pharmacological activity of the parent compound. It means having a salt according to one aspect of the invention having. The salt is formed from (1) an inorganic acid such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, or the like; Or acetic acid, propionic acid, hexanoic acid, cyclopentanepropionic acid, glycolic acid, pyruvic acid, lactic acid, malonic acid, succinic acid, malic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, 3- (4-hydroxybenzoyl) Benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, 1,2-ethane-disulfonic acid, 2-hydroxyethanesulfonic acid, benzenesulfonic acid, 4-chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid, 4-toluenesulfonic acid, camphorsulfonic acid, 4-methylbicyclo [2,2,2] -oct-2-ene-1-carboxylic acid, glucoheptonic acid, 3-phenylpropionic acid, trimethylacetic acid, tert Acid addition salts formed with organic acids such as butylacetic acid, lauryl sulfuric acid, gluconic acid, glutamic acid, hydroxynaphthoic acid, salicylic acid, stearic acid, muconic acid; Or (2) salts formed when the acidic protons present in the parent compound are substituted.
본 명세서에서 "프로드럭(prodrug)"은 어떤 약물을 화학적으로 변화시켜 물리적, 화학적 성질을 조절한 약물을 의미하며, 그 자체는 생리 활성을 나타내지 않지만 투여 후 체내에서 화학적 혹은 효소의 작용에 의해 원래의 약물로 바뀌어 약효를 발휘할 수 있다.As used herein, "prodrug" refers to a drug that modulates physical and chemical properties by chemically changing a drug, which itself does not exhibit physiological activity, but is originally produced by the action of a chemical or enzyme in the body after administration. The drug can be turned into a drug.
본 명세서에서 "수화물(hydrate)"은 물이 결합되어 있는 화합물을 의미하며, 물과 화합물 사이에 화학적인 결합력이 없는 내포 화합물을 포함하는 광범위한 개념이다. 본 명세서에서 "용매화물"은 용질의 분자나 이온과 용매의 분자나 이온 사이에 생긴 고차의 화합물을 의미한다.As used herein, "hydrate" refers to a compound to which water is bound, and is a broad concept including an inclusion compound having no chemical bonding force between water and the compound. As used herein, the term "solvate" means a higher order compound produced between molecules or ions of a solute and molecules or ions of a solvent.
일 측면에서, 본 발명은 유산균류; 및 테아닌, 이의 약학적으로 허용 가능한 염, 이의 프로드럭, 이의 수화물 또는 이의 용매화물을 포함하는, 스트레스 개선용 조성물이다. In one aspect, the present invention is lactic acid bacteria; And theanine, a pharmaceutically acceptable salt thereof, a prodrug thereof, a hydrate thereof, or a solvate thereof.
상기 테아닌(theanine)은, 아미노산의 일종으로, 바람직하게는, L-γ-글루타밀에틸아미드(glutamylethylamide) 및 N5-에틸-L-글루타민(N5-ethyl-L-glutamine)으로도 알려져 있는 L-테아닌 포함할 수 있다. The theanine (theanine) is a type of amino acid, preferably, L-γ- glutamyl ethyl amide (glutamylethylamide) and N 5 -, also known as ethyl -L- glutamine (N 5 -ethyl-L-glutamine ) L-theanine may be included.
상기 유산균은, 유산균 자체, 그 파쇄물, 그 배양산물, 또는 그 추출물을 포함하는 것일 수 있다.The lactic acid bacterium may include lactic acid bacteria itself, its lysate, its culture product, or its extract.
일 측면에서, 상기 유산균류는, 락토바실러스속 균주; 및 비피도박테리움속 균주를 포함할 수 있다. In one aspect, the lactic acid bacteria, Lactobacillus strains; And Bifidobacterium strains.
상기 락토바실러스속 균주는, 제한되지 않고, 예컨대, 락토바실러스 아시도필러스(Lactobacillus acidophilus), 락토바실러스 람노서스(Lactobacillus rhamnosus), 락토바실러스 플라타룸(Lactobacillus plantarum), 락토바실러스 카제이(Lactobacillus casei), 락토바실러스 파라카제이(Lactobacillus paracasei), 락토바실러스 루테리(Lactobacillus reuteri), 또는 락토바실러스 헬베티쿠스(Lactobacillus helveticus)를 포함할 수 있으며, 바람직하게는 락토바실러스 헬베티쿠스이다.The Lactobacillus strain is not limited to, for example, Lactobacillus acidophilus ( Lactobacillus rhamnosus ), Lactobacillus rhamnosus , Lactobacillus platarum ( Lactobacillus plantarum ), Lactobacillus casei ( Lactobacillus casei ) , Lactobacillus paracasei , Lactobacillus reuteri , or Lactobacillus helveticus , and preferably Lactobacillus helveticus .
또한, 상기 비피도박테리움속 균주는, 제한되지 않고, 비피도박테리움 롱검(Bifidobacterium longum), 비피도박테리움 브레비스(Bifidobacterium brevis), 비피도박테리움 인팬티스(Bifidobacterium infantis), 비피도박테리움 락티스(Bifidobacterium lactis), 또는, 비피도박테리움 비피듐(Bifidobacterium bifidum)을 포함할 수 있으며, 바람직하게는 비피도박테리움 롱검이다.In addition, the Bifidobacterium strain is not limited, Bifidobacterium longum ( Bifidobacterium longum ), Bifidobacterium brevis ( Bifidobacterium) brevis ), Bifidobacterium infantis ), Bifidobacterium lactis , or Bifidobacterium bifidum , preferably Bifidobacterium longgum.
상기 락토바실러스속 균주 및 비피도박테리움속 균주는 1:1 내지 20:1의 비율로 포함될 수 있다. 상기 락토바실러스속 균주 및 비피도박테리움속 균주는, 바람직하게 1:3 내지 1:17, 더욱 바람직하게 5:1 내지 16:1, 더욱 바람직하게 5:1 내지 13:1, 더욱 바람직하게 7:1 내지 13:1, 더욱 바람직하게 8:1 내지 11:1의 비율로 포함될 수 있다. 상기 락토바실러스속 균주와 비피도박테리움속 균주의 비율은, 예컨대 단위부피당 콜로니 수(CFU/ml)의 비율로 표현될 수 있다. The Lactobacillus strain and Bifidobacterium strain may be included in a ratio of 1: 1 to 20: 1. Said Lactobacillus strain and Bifidobacterium strain are preferably 1: 3 to 1:17, more preferably 5: 1 to 16: 1, more preferably 5: 1 to 13: 1, more preferably 7 It may be included in a ratio of 1: 1 to 13: 1, more preferably 8: 1 to 11: 1. The ratio of the Lactobacillus strain and the Bifidobacterium strain may be expressed by, for example, the ratio of the number of colonies (CFU / ml) per unit volume.
상기 락토바실러스속 균주와, 비피도박테리움속 균주가 상기 비율로 포함될 때 양자로부터 극대화된 효과를 얻을 수 있다.When the Lactobacillus strain and the Bifidobacterium strain are included in the ratio, it is possible to obtain the maximum effect from both.
상기 조성물로부터 얻을 수 있는 효과는 하기 중 하나 이상을 포함할 수 있다:The effect obtainable from the composition may include one or more of the following:
i) 비장세포 증식; 및 ii) 코르티코스테론(corticosterone)의 분비 억제.i) splenocyte proliferation; And ii) inhibition of secretion of corticosterone.
상기 조성물은 스트레스로 인한 신체적 증상 및 심리적 증상 중 하나 이상을 개선하는 것일 수 있다. The composition may be to ameliorate one or more of the physical and psychological symptoms caused by stress.
상기 스트레스로 인한 신체적 증상은, 소화기 질환, 신경계 질환, 비뇨기 질환, 심혈관 질환 및 피부 질환으로 구성되는 군으로부터 선택된 하나 이상을 포함할 수 있으나, 이에 제한되지 않고, 스트레스로 인해 유발되는 신체 질환이라면 종류 여부를 불문하고 포함될 수 있다.The physical symptoms caused by the stress may include one or more selected from the group consisting of digestive diseases, nervous system diseases, urinary diseases, cardiovascular diseases, and skin diseases, but are not limited thereto. It may be included with or without.
상기 소화기 질환은, 예컨대, 과민성 대장염, 소화불량, 위염, 위경련, 위십이지장궤양, 변비 등을 포함할 수 있고, 상기 신경계 질환은, 편두통을 포함할 수 있으며, 상기 비뇨기 질환은 발기부전, 불감증 등을 포함할 수 있다. The gastrointestinal disease may include, for example, irritable colitis, indigestion, gastritis, gastric cramps, gastroduodenal ulcers, constipation, and the like, the neurological disease may include migraine, and the urinary disease may include erectile dysfunction, insensitivity, etc. It may include.
또한, 상기 심혈관 질환은, 고혈압, 저혈압, 협심증 등을 포함할 수 있으며, 상기피부 질환은 탈모, 두드러기 등을 포함할 수 있다. In addition, the cardiovascular disease may include high blood pressure, low blood pressure, angina pectoris, and the like, and the skin disease may include hair loss, hives, and the like.
또한, 상기 스트레스로 인한 심리적 증상은 우울증, 불안장애, 강박증, 불면증 및 신경과민으로 구성되는 군으로부터 선택된 하나 이상을 포함할 수 있으나, 이에 제한되지 않는다.In addition, the psychological symptoms due to the stress may include one or more selected from the group consisting of depression, anxiety disorder, OCD, insomnia and nervousness, but is not limited thereto.
일 측면에서, 상기 조성물은, 화장료 조성물, 식품 조성물 또는 약학적 조성물을 포함할 수 있다.In one aspect, the composition may include a cosmetic composition, a food composition or a pharmaceutical composition.
본 발명의 실시예에 따른 화장료 조성물은 화장품학 또는 피부과학적으로 허용가능한 매질 또는 기제를 함유하여 제형화될 수 있다. 이는 국소적용에 적합한 모든 제형으로서, 현탁액, 마이크로에멀젼, 마이크로캡슐, 미세과립구 또는 이온형(리포좀) 및 비이온형의 소낭 분산제의 형태로, 또는 크림, 스킨, 로션, 파우더, 연고, 스프레이 또는 콘실 스틱의 형태로 제공될 수 있다. 또한 포말(foam)의 형태로 또는 압축된 추진제를 더 함유한 에어로졸 조성물의 형태로도 사용될 수 있다. 이들 조성물은 당해 분야의 통상적인 방법에 따라 제조될 수 있다.Cosmetic compositions according to embodiments of the present invention may be formulated containing a cosmetically or dermatologically acceptable medium or base. It is any formulation suitable for topical application, in the form of suspensions, microemulsions, microcapsules, microgranules or ionic (liposomal) and nonionic vesicle dispersants or creams, skins, lotions, powders, ointments, sprays or concealers. It may be provided in the form of a stick. It may also be used in the form of a foam or in the form of an aerosol composition further containing a compressed propellant. These compositions can be prepared according to conventional methods in the art.
또한, 화장료 조성물은 분체, 지방 물질, 유기용매, 용해제, 농축제, 겔화제, 연화제, 항산화제, 현탁화제, 안정화제, 발포제(foaming agent), 방향제, 계면활성제, 물, 이온형 또는 비이온형 유화제, 충전제, 금속이온봉쇄제, 킬레이트화제, 보존제, 비타민, 차단제, 습윤화제, 필수 오일, 염료, 안료, 친수성 또는 친유성 활성제, 지질 소낭 또는 화장품에 통상적으로 사용되는 임의의 다른 성분과 같은 화장품학 또는 피부과학 분야에서 통상적으로 사용되는 보조제를 함유할 수 있다. 상기 보조제는 화장품학 또는 피부과학 분야에서 일반적으로 사용되는 양으로 도입된다. 본 발명의 실시예들에 따른 화장료 조성물은 피부 개선 효과를 증가시키기 위하여 피부 흡수 촉진 물질을 더 함유할 수 있다.Cosmetic compositions may also contain powders, fatty substances, organic solvents, solubilizers, thickeners, gelling agents, emollients, antioxidants, suspending agents, stabilizers, foaming agents, fragrances, surfactants, water, ionic or nonionics. Such as type emulsifiers, fillers, metal ion sequestrants, chelating agents, preservatives, vitamins, blockers, wetting agents, essential oils, dyes, pigments, hydrophilic or lipophilic actives, lipid vesicles or any other ingredients commonly used in cosmetics. It may contain adjuvants conventionally used in the cosmetic or dermatology field. Such adjuvants are introduced in amounts generally used in the cosmetic or dermatological arts. The cosmetic composition according to the embodiments of the present invention may further contain a skin absorption promoting substance to increase the skin improving effect.
또한, 상기 유효성분을 포함하는 다양한 형태의 식품 조성물, 예컨대, 식품 첨가제 또는 건강 기능성 식품으로 제공될 수 있다. 상기 유효성분을 포함하는 발효유, 치즈, 요구르트, 주스, 생균제제 및 건강식품 등으로 가공될 수 있으며, 그 외 다양한 식품 첨가제의 형태로 사용될 수 있다.In addition, it can be provided in various forms of food composition, such as food additives or health functional foods containing the active ingredient. It can be processed into fermented milk, cheese, yogurt, juice, probiotic and health food, including the active ingredient, and can be used in the form of various other food additives.
상기 건강 기능성 식품은 일상 식사에서 결핍되기 쉬운 영양소나 인체에 유용한 기능을 가진 원료나 성분(기능성원료)을 사용하여 제조한 것으로, 인체의 정상적인 기능을 유지하거나 생리기능 활성화를 통하여 건강을 유지하고 개선하는 식품을 의미할 수 있으나, 이에 제한되지 않는다. The health functional food is manufactured using nutrients or ingredients (functional ingredients) having useful functions for the human body, which are easily deficient in daily meals, and maintain and improve health by maintaining normal functions of the human body or activating physiological functions. It may mean a food, but is not limited thereto.
상기 식품 조성물은 상기 유효성분을 조성물 총 중량에 대하여 0.0001 중량% 내지 80중량%로 포함될 수 있으나, 이에 제한되지 않는다.The food composition may include the active ingredient in an amount of 0.0001% to 80% by weight based on the total weight of the composition, but is not limited thereto.
상기 건강 식품용 조성물은 환제, 캅셀제, 정제, 과립제, 캬라멜제 또는 드링크제 등으로 제형화할 수 있다. 다른 구현예에서, 액제, 분말, 과립, 정제 또는 티백 등의 형태로 가공될 수도 있다.The health food composition may be formulated as pills, capsules, tablets, granules, caramels or drinks. In other embodiments, it may be processed in the form of liquids, powders, granules, tablets or tea bags and the like.
상기 조성물은 본 발명의 주 효과를 손상시키지 않는 범위 내에서 주 효과에 상승 효과를 줄 수 있는 다른 성분 등을 함유할 수 있다. 예를 들어, 물성 개선을 위하여 향료, 색소, 살균제, 산화방지제, 방부제, 보습제, 점증제, 무기염류, 유화제 및 합성 고분자 물질 등의 첨가제를 더 포함할 수 있다. 그 외에도, 수용성 비타민, 유용성 비타민, 고분자 펩티드, 고분자 다당 및 해초 엑기스 등의 보조 성분을 더 포함할 수도 있다. 상기 성분들은 제형 또는 사용 목적에 따라서 당업자가 적의 선정하여 배합할 수 있으며, 그 첨가량은 본 발명의 목적 및 효과를 손상시키지 않는 범위 내에서 선택될 수 있다. 예를 들어, 상기 성분들의 첨가량은, 조성물 전체 중량을 기준으로, 0.01 중량% 내지 10 중량%, 예를 들어 0.01중량% 내지 5중량% 일 수 있으나, 이에 제한되는 것은 아니다.The composition may contain other ingredients and the like that can give a synergistic effect to the main effect within a range that does not impair the main effect of the present invention. For example, it may further include additives such as perfumes, pigments, fungicides, antioxidants, preservatives, moisturizers, thickeners, inorganic salts, emulsifiers and synthetic polymer materials to improve physical properties. In addition, supplementary ingredients such as water soluble vitamins, oil soluble vitamins, polymer peptides, polymer polysaccharides and seaweed extract may be further included. The above ingredients may be suitably selected and formulated by those skilled in the art according to the dosage form or purpose of use, and the amount thereof may be selected within a range that does not impair the object and effect of the present invention. For example, the amount of the components added may be 0.01 wt% to 10 wt%, for example, 0.01 wt% to 5 wt%, based on the total weight of the composition, but is not limited thereto.
조성물에 있어서, 약학 조성물은 경구 또는 비경구의 여러 가지 제형일 수 있다. 제제화할 경우에는 보통 사용하는 충진제, 증량제, 결합제, 습윤제, 붕해제, 계면활성제 등의 희석제 또는 부형제를 사용하여 조제된다. 경구투여를 위한 고형제제에는 정제, 환제, 산제, 과립제, 연질 또는 경질 캡슐제 등이 포함되며, 이러한 고형제제는 하나 이상의 화합물에 적어도 하나 이상의 부형제 예를 들면, 전분, 탄산칼슘, 수크로오스(sucrose) 또는 락토오스(lactose), 젤라틴 등을 섞어 조제된다. 또한 단순한 부형제 이외에 스테아린산 마그네슘, 탈크 등과 같은 윤활제들도 사용된다. 경구투여를 위한 액상제제로는 현탁제, 내용액제, 유제, 시럽제 등이 해당되는데 흔히 사용되는 단순 희석제인 물, 리퀴드 파라핀 이외에 여러 가지 부형제, 예를 들면 습윤제, 감미제, 방향제, 보존제 등이 포함될 수 있다. 비경구투여를 위한 제제에는 멸균된 수용액, 비수성용제, 현탁제, 유제, 동결건조제제, 좌제가 포함된다. 비수성용제, 현탁용제로는 프로필렌글리콜(propylene glycol), 폴리에틸렌 글리콜, 올리브 오일과 같은 식물성 기름, 에틸올레이트와 같은 주사 가능한 에스테르 등이 사용될 수 있다. 좌제의 기제로는 위텝솔(witepsol), 마크로골, 트윈(tween) 61, 카카오지, 라우린지, 글리세로젤라틴 등이 사용될 수 있다.In the composition, the pharmaceutical composition may be in various formulations, oral or parenteral. When formulated, diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents, and surfactants are usually used. Solid form preparations for oral administration include tablets, pills, powders, granules, soft or hard capsules, and the like, which may contain at least one excipient such as starch, calcium carbonate, sucrose, or the like. Or lactose, gelatin, or the like is mixed. In addition to simple excipients, lubricants such as magnesium stearate, talc and the like are also used. Liquid preparations for oral administration include suspensions, liquid solutions, emulsions, and syrups, and various excipients such as wetting agents, sweeteners, fragrances, and preservatives, in addition to commonly used simple diluents such as water and liquid paraffin, may be included. have. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, suppositories. As the non-aqueous solvent and the suspension solvent, propylene glycol, polyethylene glycol, vegetable oils such as olive oil, injectable esters such as ethyl oleate, and the like can be used. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerogelatin and the like can be used.
본 발명의 일 실시예에 따른 약학 조성물의 유효 성분은 대상의 연령, 성별, 체중, 병리 상태 및 그 심각도, 투여 경로 또는 처방자의 판단에 따라 달라질 것이나, 하기 기재된 투여량으로 투여하는 것이 바람직하다.The active ingredient of the pharmaceutical composition according to an embodiment of the present invention will vary depending on the age, sex, weight, pathology and severity of the subject, the route of administration, or the judgment of the prescriber, but is preferably administered at the dosages described below.
본 발명의 일 관점에 따른 조성물은, 쥐, 생쥐, 가축, 인간 등의 포유동물에 비경구, 경구 등의 다양한 경로로 투여될 수 있으며, 투여의 모든 방식은 예상될 수 있는데, 예를 들면, 경구, 경피(trandermally), 정맥, 근육, 피하주사에 의해 투여될 수 있다.The composition according to one aspect of the present invention may be administered to mammals such as rats, mice, livestock, humans by various routes, such as parenteral, oral, and all modes of administration may be expected. Oral, transdermally, intravenous, intramuscular, subcutaneous injection.
일 측면에서, 본 발명은 상기 조성물; 및 지시서를 포함하며, 상기 지시서는, 유산균류는 108~1010CFU/day로 투여하고, 테아닌은 5~500mg/kg/day로 투여하는 내용을 포함하는, 스트레스 개선용 키트이다.In one aspect, the present invention is a composition; And instructions, wherein the instructions, lactic acid bacteria are administered in 10 8 ~ 10 10 CFU / day, theanine is a kit for stress improvement, comprising the contents administered at 5 ~ 500mg / kg / day.
본 명세서에서 "키트"는, 물품(article)을 의미할 수 있으며, 예컨대, 상기 키트 또는 물품은 화장품, 식품, 또는 약품을 포함할 수 있다.As used herein, "kit" may mean an article, for example, the kit or article may include cosmetics, food, or drugs.
상기 유산균류는, 바람직하게는 108~0.8X1010CFU/day로 투여될 수 있고, 더욱 바람직하게는 2X108~0.7X1010 로 투여될 수 있다.The lactic acid bacteria, preferably may be administered at 10 8 ~ 0.8X10 10 CFU / day, more preferably may be administered at 2X10 8 ~ 0.7X10 10 .
또한 상기 지시서는 유산균류는, 락토바실러스 헬베티쿠스(Lactobacillus helveticus); 및 비피도박테리움 롱검(Bifidobacterium longum)을 포함하고, 상기 락토바실러스 헬베티쿠스와 비피도박테리움 롱검은, 순서대로 5X107~5X109 CFU/day, 1X107~1X109 CFU/day로 투여하는 내용을 포함할 수 있다.In addition, the instructions are lactic acid bacteria, Lactobacillus helveticus ( Lactobacillus helveticus ); And including Bifidobacterium ronggeom (Bifidobacterium longum), and the Lactobacillus helveticus and Bifidobacterium long black, in order to 5X10 7 ~ 5X10 9 CFU / day , 1X10 7 ~ administering to 1X10 9 CFU / day May contain content.
바람직하게, 상기 락토바실러스 헬베티쿠스와 비피도박테리움 롱검은, 순서대로 108~5X109 CFU/day, 2X107~1X109 CFU/day로 투여될 수 있으며, 더욱 바람직하게, 2X108~5X109 CFU/day, 2X107~7X108 CFU/day로 투여될 수 있다.Preferably, the Lactobacillus helveticus and Bifidobacterium long black, in order to 10 8 ~ 5X10 9 CFU / day , 2X10 7 ~ may be administered to 1X10 9 CFU / day, more preferably, 2X10 8 ~ 5X10 9 CFU / day, 2 × 10 7 to 7 × 10 8 CFU / day.
또한, 상기 테아닌의 투여량은, 바람직하게 10~450mg/kg/day, 더욱 바람직하게는 40~450 mg/kg/day, 더욱 바람직하게, 80~450 mg/kg/day일 수 있다.In addition, the dosage of theanine is preferably 10 to 450 mg / kg / day, more preferably 40 to 450 mg / kg / day, and more preferably 80 to 450 mg / kg / day.
이하, 실시예를 통하여 본 발명을 더욱 상세히 설명하고자 한다. 이들 실시예는 오로지 본 발명을 예시하기 위한 것으로서, 본 발명의 범위가 이들 실시예에 의해 제한되는 것으로 해석되지 않는 것은 당업계에서 통상의 지식을 가진 자에게 있어서 자명할 것이다.Hereinafter, the present invention will be described in more detail with reference to Examples. These examples are only for illustrating the present invention, it will be apparent to those skilled in the art that the scope of the present invention is not to be construed as limited by these examples.
[실험예 1] 마우스 비장세포 증식능 평가Experimental Example 1 Evaluation of Mouse Splenocyte Proliferation
본 발명의 락토바실러스 헬베티쿠스 R52 (Lactobacillus helveticus Rosell-52)을 MRS 액체배지 (Difco, USA)에서 37℃, 20시간 동안 배양하였다. 균체를 원심분리한 후 PBS로 2~3번 세척한 후 남아있는 배지성분을 제거하였다. 연속희석법 (serial dilution)의 과정을 거쳐 균수를 측정하여 균체의 농도가 1~5X109 CFU/mL이 되도록 PBS에 희석한 후 시험에 사용하였다. Lactobacillus helveticus of Lactobacillus helveticus R52 of the present invention Rosell-52) was incubated for 20 hours at 37 ℃ in MRS liquid medium (Difco, USA). After centrifugation of the cells, the remaining medium components were removed after washing with PBS 2-3 times. The number of bacteria was measured through serial dilution and diluted in PBS so that the concentration of the cells was 1-5 × 10 9 CFU / mL.
본 발명의 비피도박테리움 롱검 R175 (Bifidobacterium longum Rosell-175)을 BL 액체배지 (Difco, USA)에서 혐기조건 하에서 37℃, 20시간 동안 배양하였다. 균체를 원심분리한 후 PBS로 2~3번 세척한 후 남아있는 배지성분을 제거하였다. 연속희석법 (serial dilution)의 과정을 거쳐 균수를 측정하여 균체의 농도가 1~5X109 CFU/mL이 되도록 PBS에 희석한 후 시험에 사용하였다. Bifidobacterium longgum R175 of the present invention ( Bifidobacterium longum Rosell-175) was incubated in BL liquid medium (Difco, USA) for 20 hours at 37 ℃ under anaerobic conditions. After centrifugation of the cells, the remaining medium components were removed after washing with PBS 2-3 times. The number of bacteria was measured through serial dilution and diluted in PBS so that the concentration of the cells was 1-5 × 10 9 CFU / mL.
상기 균주들은, Lallemand Health Solutions Inc. (Mirabel QC, Canada)로부터 구입한 것을 사용하였다.The strains were Lallemand Health Solutions Inc. (Mirabel QC, Canada) was used.
실험동물은 20g 전후의 6주령 암컷 마우스 (C57BL/6)을 사용하였으며, 명암주기 12시간, 온도 25±5℃, 습도60±10%의 조건하에서 사육하였다. 스트레스 유발을 위해 마우스는 50 mL튜브에 사료 및 음수 급여없이 17시간 억제스트레스를 유발시켰다 (Iwakabe K, Shimada M, Ohta A, Yahata T, Ohmi Y, Habu S, Nishimura T. 1998. The restraint stress drives a shift in Th1/Th2 balance toward Th2-dominant immunity in mice. Immuno Letter 62:39-43).Experimental animals were used 6 weeks old female mice (C57BL / 6) around 20g, and were bred under the conditions of 12 hours contrast cycle, temperature 25 ± 5 ℃,
스트레스 유도 마우스의 비장을 무균적으로 적출하고 비장세포를 분리한 후 RPMI 1640배지에 10% FBS (fetal bovine serum)을 넣고, 이를 이용하여 1X107 cell/mL로 현탁하여 96 well 플레이트에 분주하였다. 각 well에 락토바실러스 헬베티쿠스 R52 (Lactobacillus helveticus Rosell-52)및 비피도박테리움 롱검 R175 (Bifidobacterium longum Rosell-175)를 희석배수에 따라 10% FBS-RPMI 1640 배지액에 현탁시켜 well 당 20㎕씩 가하였다. 여기에 Concanavalin A (ConA；2㎍/ml) 또는 10% FBS-RPMI 1640 배지액을 20㎕ 가한 후 72시간동안 배양(37℃, 5% CO2)하였다. 비장세포의 증식정도는 생존세포의 효소작용에 의해 MTT 시약이 환원되어 생성되는 formazan의 양을 측정함으로써 평가하는 MTT assay를 이용하여 측정하였다. 즉, 배양 후 각 well에 MTT용액(5 mg/ml)을 50㎕ 씩 (250㎍/ml) 가하고 4시간 동안 배양한 후에 원심분리(400 rpm, 5 min, 4℃)하여 상등액을 제거하였다. Dimethyl sulfoxide (DMSO)원액을 well당 50㎕씩 가하고 10분 이내에 ELISA reader로 흡광도(570 nm)를 측정하였다. Spleens of stress-induced mice were aseptically extracted and splenocytes were isolated, and 10% FBS (fetal bovine serum) was added to RPMI 1640 medium, which was suspended at 1 × 10 7 cells / mL and dispensed into 96 well plates. Lactobacillus helveticus in each well Lactobacillus helveticus Rosell-52) and Bifidobacterium longum R175 Rosell-175) was suspended in 10% FBS-RPMI 1640 medium according to the dilution factor and added to 20 μl per well. 20 μl of Concanavalin A (ConA; 2 μg / ml) or 10% FBS-RPMI 1640 medium was added thereto, followed by incubation for 72 hours (37 ° C., 5% CO 2). The proliferation of splenocytes was measured by MTT assay, which was evaluated by measuring the amount of formazan produced by reducing MTT reagent by enzymatic action of viable cells. That is, after incubation, 50 μl of MTT solution (5 mg / ml) was added to each well (250 μg / ml), followed by incubation for 4 hours, followed by centrifugation (400 rpm, 5 min, 4 ° C.) to remove the supernatant. Dimethyl sulfoxide (DMSO) stock solution was added 50 µl per well and absorbance (570 nm) was measured by ELISA reader within 10 minutes.
스트레스유도 마우스의 비장세포는 정상 마우스에서 분리한 비장세포를 ConA로 자극한 경우 나타내는 증식능보다 30% 감소하는 경향을 나타내었다. 락토바실러스 헬베티쿠스 R52 및 비피도박테리움 롱검 R175의 비율에 따라 증식능 (+, 0~10; ++, 10~20; +++, 20~30; ++++, >30)을 평가하였다. Splenocytes of stress-induced mice showed a 30% reduction in the proliferative capacity of splenocytes isolated from normal mice when stimulated with ConA. The proliferative capacity (+, 0-10; ++, 10-20; +++, 20-30; ++++,> 30) was evaluated according to the ratio of Lactobacillus helvetticus R52 and Bifidobacterium longgum R175. It was.
그 결과, 락토바실러스 헬베티쿠스와, 비피도박테리움 롱검을 약 3:1~15:1의 비율로 혼합하였을 때, 스트레스에 노출된 비장세포의 증식 효과가 우수함을 확인할 수 있었다(표 1).As a result, when Lactobacillus helvetticus and Bifidobacterium longgum were mixed at a ratio of about 3: 1 to 15: 1, it was confirmed that the proliferative effect of splenocytes exposed to stress was excellent (Table 1). .
[실험예 2] 스트레스 유도 마우스의 혈중 코르티코스테론 농도의 변화Experimental Example 2 Changes in Serum Corticosterone Levels in Stress-induced Mice
실험동물은 20g 전후의 6주령 암컷 마우스 (C57BL/6)을 사용하였으며, 명암주기 12시간, 온도 25±5℃, 습도60±10%의 조건하에서 사육하였다. 비교예 1~6 및 실시예 1~4를 마우스 전용사료 (AIN-93 M diet, maintenance formulation, Korea)에 혼합하여 1주일간 급여하였다. 스트레스 유도는 50 mL튜브에 사료 및 음수 급여없이 17시간 동안 물리적으로 고정하여 스트레스를 유발하였다 (Iwakabe K, Shimada M, Ohta A, Yahata T, Ohmi Y, Habu S, Nishimura T. 1998. The restraint stress drives a shift in Th1/Th2 balance toward Th2-dominant immunity in mice. Immuno Letter 62:39-43).Experimental animals were used 6 weeks old female mice (C57BL / 6) around 20g, and were bred under the conditions of 12 hours contrast cycle, temperature 25 ± 5 ℃,
마우스 용량은 마우스의 일일 평균사료섭취량 (2~3g)을 사람에 대한 용량으로의 환산하여 나타내었다.인간등가용량 (HED) 은 체표면적을 기준으로 animal dose (in mg/kg) = HED (human equivalent dose, in mg/kg, 60 kg 기준)/BSA-CF(body surface area dose conversion factor)의 계산식(Guidance for Industry and Reviewers - Estimating the Safe Starting Dose in Clinical Trials for Therapeutics in Adult Healthy Volunteers: Table 3) Conversion of Animal Doses to Human Equivalent Doses (HED) Based on Body Surface Area 등 참조)으로 산출하였다.The mouse dose is expressed as the average daily food intake of the mouse (2-3 g) in terms of the human dose. The equivalent human dose (HED) is the animal dose (in mg / kg) = HED (human) based on the body surface area. Guidance for Industry and Reviewers-Estimating the Safe Starting Dose in Clinical Trials for Therapeutics in Adult Healthy Volunteers: Table 3 (Equivalent dose, in mg / kg, 60 kg) / BSA-CF ) Conversion of Animal Doses to Human Equivalent Doses (HED) Based on Body Surface Area, etc.).
스트레스 유도 후 마우스 혈액에서 혈중 코르티코스테론 농도를 ELISA kit (MP Biomedicals, CA, USA)를 이용하여 측정하였다.After stress induction, blood corticosteroid concentrations in mouse blood were measured using an ELISA kit (MP Biomedicals, CA, USA).
상기 실험에 사용된 조성물들의 조성과, 조성물을 이용한 실험 결과는 하기 표 2 및 표 3에 기재된 바와 같다. The composition of the compositions used in the above experiments and the results of the experiments using the compositions are as described in Tables 2 and 3 below.
그 결과, 상기 표 3에서 볼 수 있듯이, L-테아닌와 유산균을 병용할 경우, L-테아닌 또는 유산균 각각을 사용할 경우에 비하여, 현저히 우수한 혈중 코르티코스테론 저감 효과가 있음을 알 수 있었고, 특히, 락토바실러스 헬베티쿠스와 비피도박테리움 롱검이 약 9:1로 포함될 경우 가장 우수한 코르티코스테론 저감 효과를 확인할 수 있었다.As a result, as shown in Table 3, when L- theanine and lactic acid bacteria used in combination, it was found that there is a remarkably excellent effect of reducing blood corticosteroids, especially when using L- theanine or lactic acid bacteria, respectively, lactose When Bacillus helveticus and Bifidobacterium longgum were included as about 9: 1, the most effective corticosteroid reduction was confirmed.
[실험예 3] 스트레스 유도 마우스의 혈중 사이토카인 농도의 변화Experimental Example 3 Changes in Blood Cytokine Concentrations in Stress-induced Mice
스트레스 유도 마우스의 비장을 무균적으로 적출하고 비장세포를 분리한 후 RPMI 1640배지에 10% FBS (fetal bovine serum)을 넣고, 이를 이용하여 1X107 cell/mL로 현탁하여 96 well 플레이트에 분주하였다. 각 well에 락토바실러스 헬버티쿠스 R52 및 비피도박테리움 롱검 R175를 희석배수에 따라 10% FBS-RPMI 1640 배지액에 현탁시켜 well 당 20㎕씩 가하였다. 여기에 Concanavalin A (ConA；2㎍/ml) 또는 10% FBS-RPMI 1640 배지액을 20㎕ 가한 후 72시간동안 배양 (37℃, 5% CO2)하였다. 배양 종료 후, 배양상층액에 존재하는 IFN-γ 및 IL-4의 양은 각 사이토카인에 대한 ELISA kit (BD Biosciences, USA)를 사용하여 분석하였다.Spleens of stress-induced mice were aseptically extracted and splenocytes were isolated, and 10% FBS (fetal bovine serum) was added to RPMI 1640 medium, which was suspended at 1 × 10 7 cells / mL and dispensed into 96 well plates. To each well, Lactobacillus helticus R52 and Bifidobacterium longgum R175 were suspended in 10% FBS-RPMI 1640 medium according to the dilution factor, and 20 μl per well was added. 20 μl of Concanavalin A (ConA; 2 μg / ml) or 10% FBS-RPMI 1640 medium was added thereto, followed by incubation for 72 hours (37 ° C., 5% CO 2). After incubation, the amount of IFN-γ and IL-4 present in the culture supernatant was analyzed using an ELISA kit (BD Biosciences, USA) for each cytokine.
그 결과, L-테아닌과 유산균을 병용투여했을 경우에, 그렇지 않은 경우에 비하여 스트레스 유도 마우스의 사이토카인 분비량이 더 높음을 확인할 수 있었다(표 4 및 도 2 참조).As a result, when L-theanine and lactic acid bacteria were co-administered, it was confirmed that the cytokine secretion of stress-induced mice was higher than that of the other cases (see Table 4 and FIG. 2).
[실험예 4] 십자형 미로 검사 (Elevated plus-maze test)Experimental Example 4 Elevated plus-maze test
본 발명에 따른 혼합물의 불안감소 또는 억제효과를 확인하기 위하여, 실험예 2의 결과를 바탕으로 L-테아닌 및 유산균 혼합물을 하기와 같이 십자형 높은 미로검사를 통하여 평가하였다.In order to confirm anxiety reduction or inhibitory effect of the mixture according to the present invention, the L-theanine and lactic acid bacteria mixture was evaluated through a crosswise high maze test as follows based on the results of Experimental Example 2.
실험동물은 20g 전후의 6주령 암컷 마우스 (C57BL/6)을 사용하였으며, 명암주기 12시간, 온도 25±5℃, 습도60±10%의 조건하에서 사육하였다. L-테아닌 및 유산균 혼합물은 마우스 전용사료 (AIN-93 M diet, maintenance formulation, Korea)에 혼합하여 4주일간 급여하였다. 이때 L-테아닌의 농도는 200 mg/day 용량, 락토바실러스 헬버티쿠스 R52는 2.7X109 CFU/day 용량, 비피도박테리움 롱검 R175는 0.3X109 CFU/day 용량으로 투여하였다.Experimental animals were used 6 weeks old female mice (C57BL / 6) around 20g, and were bred under the conditions of 12 hours contrast cycle, temperature 25 ± 5 ℃,
마우스에 시험물질의 급여를 종료한 후, 마우스를 십자로 시험장치의 중앙에 위치시켜 불안증을 유발하게 만든 후, 상기 장치위에서 마우스의 행동양상 변화를 관찰하여 혼합물의 불안감소 또는 억제를 평가하였다. After the feeding of the test substance to the mouse, the mouse was placed crosswise at the center of the test apparatus to cause anxiety, and the change in behavior of the mouse was observed on the apparatus to evaluate anxiety reduction or inhibition of the mixture.
십자형 미로는 아크릴로 제작되며, 바닥으로부터 40cm 떨어진 곳에 십자형(+)의 미로가 구성된다. 상기 미로의 통로 4개 중 마주보는 2개의 통로는 개방(open arm) 되어 있고, 또 다른 마주보는 2개의 통로(closed arm)는 높이 20cm의 벽으로 둘러싸여 있다. 중앙의 플랫폼은 가로와 세로의 길이가 각각 8cm이며, 실험 시작 시 마우스를 중앙 플랫폼에 두고 5분 동안 미로를 자유롭게 탐색하도록 하였다. 그 후, 마우스가 개방된 통로(open arm)와 폐쇄된 통로(closed arm)에 머문 시간을 분석하였다. 양성 대조군은 행동약리 시험 전 30분전에 Diazepam을 2 mg/kg 용량으로 복강투여하였으며, 음성 대조군으로는 시험물질을 투여하지 않은 마우스를 사용하였다. The cruciform maze is made of acrylic and consists of a cruciform (+)
EPM은 흔히 불안 완화제나 불안유도 약물의 효과를 확인하는데 사용되는 모델로 쥐가 좁고 어두운 곳을 선호하는 습성을 이용한 것이다. 벽이 설치된 closed arm보다 벽이 없는 open arm에서 더 많은 불안을 느끼므로 불안이 많을수록 open arm에 머무는 시간이 짧아진다. 항불안 약물의 검색은 open arm에서 머문 시간의 연장 및 open arm으로의 진입 횟수의 증가로 평가되는데 이는 close arm 으로의 진입 횟수 및 머문 시간의 감소로 표현된다. 본 발명에서 EPM을 이용한 불안 실험동물 모델에서 L-테아닌 및 유산균 혼합물의 경우 open arm에 머문 시간 및 open arm으로의 진입 횟수가 대조군에 비하여 증가됨을 관찰할 수 있었다. L-테아닌 및 유산균 혼합물 단독 투여군에 비해 L-테아닌과 유산균 혼합물의 병용투여군에서 open arm에 머무르는 시간 및 진입횟수가 증가하였다. EPM is a model commonly used to identify the effects of anxiolytics or anxiety-inducing drugs, using rats' preference for narrow, dark areas. The more anxiety the shorter the staying time on the open arm is, because more anxiety is felt in the open arm without the wall than in the closed arm. The detection of anti-anxiety drugs is assessed by the extension of the time spent in the open arm and the increase in the number of entry into the open arm, which is expressed by the decrease in the number of entry into the close arm and the length of stay. In the present invention, the anxiety test animal model using the EPM L- theanine and lactic acid bacteria mixture was observed that the time to stay in the open arm and the number of entry into the open arm was increased compared to the control group. Compared to the L-theanine and lactic acid bacterial mixture alone group, the combined arm and L-theanine and lactic acid bacterial mixture group stayed in the open arm and the number of entry times increased.
이러한 시험결과로부터, 본 발명에 따라 L-테아닌과 유산균 혼합물(락토바실러스 헬버티쿠스 R52 및 비피도박테리움 롱검 R175의 혼합물)을 유효 성분으로 함유하는 조성물을 이용할 경우, 시험 동물의 불안증을 감소시킬 수 있다는 것을 확인할 수 있었다(도 3).From these test results, according to the present invention, when using a composition containing a mixture of L- theanine and lactic acid bacteria (mixture of Lactobacillus helticus R52 and Bifidobacterium long gum R175) as an active ingredient, anxiety in test animals can be reduced. It could be confirmed that it can (Fig. 3).
Claims (11)
테아닌, 이의 약학적으로 허용 가능한 염, 이의 프로드럭, 이의 수화물 또는 이의 용매화물을 포함하는, 스트레스 개선용 조성물.
Lactic acid bacteria; And
Theanine, a pharmaceutically acceptable salt thereof, a prodrug thereof, a hydrate thereof, or a solvate thereof, the composition for improving stress.
상기 유산균류는,
락토바실러스속 균주; 및 비피도박테리움속 균주를 포함하는, 스트레스 개선용 조성물.
The method of claim 1,
The lactic acid bacteria,
Lactobacillus strains; And Bifidobacterium spp. Strain, the composition for improving stress.
상기 조성물은,
스트레스로 인한, 신체적 증상; 및 심리적 증상 중 하나 이상을 개선하는, 스트레스 개선용 조성물.
The method of claim 1,
The composition,
Physical symptoms due to stress; And improving at least one of psychological symptoms.
상기 락토바실러스속 균주 및 비피도박테리움속 균주는,
1:1 내지 20:1의 비율로 포함되는, 스트레스 개선용 조성물.
The method of claim 1,
The Lactobacillus strain and Bifidobacterium strain,
Included in a ratio of 1: 1 to 20: 1, the composition for improving stress.
제2항에 있어서,
상기 락토바실러스속 균주는 락토바실러스 헬베티쿠스(Lactobacillus helveticus)를 포함하고,
상기 비피도박테리움속 균주는 비피도박테리움 롱검(Bifidobacterium longum)을 포함하는, 스트레스 개선용 조성물.
The method of claim 2,
The Lactobacillus strain comprises Lactobacillus helveticus , Lactobacillus helveticus ,
The Bifidobacterium strain comprises a Bifidobacterium longum ( Bifidobacterium longum ), a composition for improving stress.
상기 조성물은, 하기 중 하나 이상의 특징을 포함하는, 스트레스 개선용 조성물:
i) 비장세포 증식 촉진; 및
ii) 코르티코스테론(corticosterone)의 분비 억제.
The method of claim 1,
The composition, stress improvement composition comprising one or more of the following characteristics:
i) promoting splenocyte proliferation; And
ii) inhibition of secretion of corticosterone.
상기 스트레스로 인한 신체적 증상은, 소화기 질환, 신경계 질환, 비뇨기 질환, 심혈관 질환 및 피부 질환으로 구성되는 군으로부터 선택된 하나 이상을 포함하고,
상기 스트레스로 인한 심리적 증상은, 우울증, 불안장애, 불면증 및 신경과민으로 구성되는 군으로부터 선택된 하나 이상을 포함하는, 스트레스 개선용 조성물.
The method of claim 3,
Physical symptoms caused by the stress include one or more selected from the group consisting of digestive diseases, nervous system diseases, urinary diseases, cardiovascular diseases and skin diseases,
The psychological symptoms due to the stress, depression, anxiety disorders, insomnia and neurological hypersensitivity comprising at least one selected from the group consisting of, stress improvement composition.
상기 테아닌은, L-테아닌을 포함하는, 스트레스 개선용 조성물.
The method of claim 1,
The theanine, L- theanine, the composition for stress improvement.
상기 조성물은,
화장료 조성물, 식품 조성물 또는 약학적 조성물을 포함하는, 스트레스 개선용 조성물.
The method of claim 1,
The composition,
A composition for improving stress, comprising a cosmetic composition, a food composition or a pharmaceutical composition.
상기 지시서는,
유산균류는 108~1010CFU/day로 투여하고,
테아닌은 5~500mg/kg/day로 투여하는 내용을 포함하는, 스트레스 개선용 키트.
The composition of any one of claims 1 to 9; And instructions,
The above instructions,
Lactobacillus is administered at 10 8 ~ 10 10 CFU / day,
Theanine is a kit for stress improvement comprising the contents administered at 5 ~ 500mg / kg / day.
상기 지시서는,
유산균류는, 락토바실러스 헬베티쿠스(Lactobacillus helveticus); 및 비피도박테리움 롱검(Bifidobacterium longum)을 포함하고,
상기 락토바실러스 헬베티쿠스와 비피도박테리움 롱검은,
순서대로 5X107~5X109 CFU/day, 1X107~1X109 CFU/day로 투여하는 내용을 포함하는, 스트레스 개선용 키트.
The method of claim 10,
The above instructions,
Lactobacillus, Lactobacillus helveticus ( Lactobacillus helveticus ); And Bifidobacterium longum ,
The Lactobacillus Helveticus and Bifidobacterium long black,
Order of 5X10 7 ~ 5X10 9 CFU / day , 1X10 7 ~ kit for stress improvement, including the contents of the administration to 1X10 9 CFU / day.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| KR20240144540A (en) | 2023-03-22 | 2024-10-02 | 경남대학교 산학협력단 | Composition for ameliorating stress containing extracts of Valeriana fauriel Briquet, Scutellaria baicalensis Georgi, citrus unshiu peel, Matricaria chamomilla, wormwood and Prunella Vulgaris L. lilacina Nakai |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20140043544A (en) | 2012-09-24 | 2014-04-10 | 재단법인 경기과학기술진흥원 | Ant-stress composition using an extract of xylosma congesta |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| KR20140043544A (en) | 2012-09-24 | 2014-04-10 | 재단법인 경기과학기술진흥원 | Ant-stress composition using an extract of xylosma congesta |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20240144540A (en) | 2023-03-22 | 2024-10-02 | 경남대학교 산학협력단 | Composition for ameliorating stress containing extracts of Valeriana fauriel Briquet, Scutellaria baicalensis Georgi, citrus unshiu peel, Matricaria chamomilla, wormwood and Prunella Vulgaris L. lilacina Nakai |
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