KR101779536B1 - Composition for treating sleep disorder and enhacning sleeping time containing polygonatum extract - Google Patents
Composition for treating sleep disorder and enhacning sleeping time containing polygonatum extract Download PDFInfo
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- KR101779536B1 KR101779536B1 KR1020170058492A KR20170058492A KR101779536B1 KR 101779536 B1 KR101779536 B1 KR 101779536B1 KR 1020170058492 A KR1020170058492 A KR 1020170058492A KR 20170058492 A KR20170058492 A KR 20170058492A KR 101779536 B1 KR101779536 B1 KR 101779536B1
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- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/88—Liliopsida (monocotyledons)
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- A—HUMAN NECESSITIES
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- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/105—Plant extracts, their artificial duplicates or their derivatives
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2200/00—Function of food ingredients
- A23V2200/30—Foods, ingredients or supplements having a functional effect on health
- A23V2200/322—Foods, ingredients or supplements having a functional effect on health having an effect on the health of the nervous system or on mental function
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- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Medicines Containing Plant Substances (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
Abstract
Description
본 발명은 식물 추출물을 유효 성분으로 함유하는 조성물에 관한 것으로, 더욱 상세하게는 둥굴레 추출물을 유효 성분으로 함유하는 수면 장애 치료용 약학 조성물 및/또는 의약품 및/또는 수면 시간 증진 및/또는 수면 질 개선을 위한 조성물 또는 기능성 식품에 관한 것이다.The present invention relates to a composition containing a plant extract as an active ingredient, and more particularly, to a pharmaceutical composition and / or medicament for treating sleep disorders containing an extract of Aspergillus oryzae as an active ingredient and / or a composition for improving sleep time and / ≪ / RTI >
적절한 수면은 다음날 정상적인 활동을 하기 위하여 몸과 마음의 피로를 회복시키기 위해 반드시 필요하다. 최근의 연구 결과에 따르면, 적절한 수면을 통하여 집중력, 기억력이 향상될 뿐만 아니라, 비만 및 당뇨를 예방하며 졸음운전 등의 안전사고 예방 등과 관련해서도 적절한 수면은 밀접한 관련이 있다. 따라서 불면증 등의 수면장애 개선 및 수면의 질 향상은 곧 삶의 질 향상으로 이어질 수 있는 매우 중요한 요소이다. Proper sleep is essential to restore body and mind fatigue for normal activities the next day. Recent research has shown that adequate sleep does not only increase concentration and memory, but also prevent obesity and diabetes and prevent accidents such as drowsiness. Therefore, improvement of sleep disorder such as insomnia and improvement of sleep quality are very important factors that can lead to improvement of quality of life.
그러나 현대사회에서 많은 사람들은 연장 업무나 야간 학습으로 인하여 물리적인 수면 시간이 부족해지거나 스트레스, 불안, 초조와 같은 심리적 압박감으로 인하여 수면 장애를 겪으면서 충분한 수면을 취하지 못하고 있다. 통계에 의하면 약 15%에 가까운 사람들이 불안증상으로 인하여 불면증을 겪고 있어 약물 치료가 필요하다고 알려져 있고, 나이가 들수록 불면증 환자가 증가하고 있다고 알려져 있다. 불면증과 같은 수면 장애를 치료하는 약물로는 벤조다이아제핀 계열과 세로토닌 계열 등의 약물이 사용되고 있으나, 이러한 약물들을 장기간 복용할 때 약물에 대한 내성 및 약물 의존성이 형성되는 부작용이 발생하여 초기 또는 일시적인 불면을 호소하는 사람들에게 사용하기에는 부적합한 면이 있다. In modern society, however, many people are unable to sleep adequately due to lack of physical sleeping hours due to extended work or nighttime learning, or due to psychological pressure such as stress, anxiety, and agitation, resulting in sleep disorders. Statistics show that close to 15% of people are experiencing insomnia due to anxiety symptoms, so it is known that medication is needed and that the older people are increasing the insomnia. Drugs such as benzodiazepines and serotonines have been used to treat sleep disorders such as insomnia. However, long-term use of these drugs has resulted in the adverse effects of drug resistance and drug dependence, There is something unsuitable for use by those who appeal.
따라서, 수면제를 장기 복용해야 하는 사람들의 경우 이를 대체할 수 있는 수단으로 천연 소재에서 유래한 수면 보조 및/또는 수면 질 개선제에 대한 필요성이 증가하고 있다. 특히, 선진국에서는 종래 화학 합성에 의해 제조되는 수면제를 대체하거나 종래 수면제의 부작용을 회피할 수 있는 수면 증진 건강기능식품에 대한 관심이 높아지고 있다. 이처럼, 개인의 삶의 질에 대한 관심과 수면에 대한 중요성이 증가함에 따라 수면의 질을 개선하고자 하는 욕구가 높아지고 있어 불면증과 같은 수면 장애를 치료 또는 예방할 수 있는 의약품뿐만 아니라 일반인의 수면의 질을 향상시킬 수 있는 건강기능식품에 대한 수요도 더 커질 것으로 전망된다. Thus, there is an increasing need for sleep aids and / or sleep quality improvers derived from natural materials as a means of replacing those who need long-term use of sleeping pills. Particularly in developed countries, there is increasing interest in replacing sleeping pills manufactured by conventional chemical synthesis or sleep-improving health functional foods that can avoid side effects of conventional sleeping pills. Thus, as interest in the quality of life of individuals increases and the importance of sleep increases, the desire to improve the quality of sleep is increasing. Therefore, not only drugs that can treat or prevent sleep disorders such as insomnia, The demand for health functional foods that can be improved is expected to increase.
현대 한의학에서 수면장애는 불안 및 신경과민과 관련된 질환으로 분류하고 있으며, 한의학적 접근을 통한 수면장애 연구가 다각적으로 보고되고 있다. 한방에서 신경과민에 관한 연구로는 대표적으로 산조인, 길초근 등이 있다. 이러한 한약재는 저-용량에서 항-불안, 수면보고 효과를 나타내고, 고-용량에서는 진정작용, 항-우울 효과를 가지는 것으로 알려져 있다. 그 밖에 항-불안 생리활성 효능을 갖는 약재로는 황금, 현삼 및 천마 등이 있다. In Hyundai Oriental medicine, sleep disorders are classified as diseases related to anxiety and nervous irritation, and studies of sleep disorders through a oriental medicine approach are reported in various ways. There are many studies on neuropsychiatric disorders in Oriental medicine. These herbal medicines are known to have anti-anxiety and sleep reporting effects at low-dose, and sedative and anti-depressive effects at high-dose. Other medicinal products with anti-anxiolytic activity include gold, hansam, and horse chestnut.
한방에서 사용되는 천연 약재나 생약을 응용한 수면 장애를 치료 또는 수면 개선을 위한 연구가 활발하게 진행되었다. 예를 들어, 대한민국공개특허 제2007-0070307호에서는 항히스타민 약에 다양한 생약 성분을 포함하는 수면 개선 의약 조성물을 제안하고 있다. 또한, 대한민국특허 제1020245호에서는 감초 에탄올 추출물 및/또는 합환피 에탄올 추출물로 이루어진 수면 개선용 조성물을 개시하고 있다. Research has been actively conducted to treat sleep disorder or to improve sleep by applying natural herbal medicine or herbal medicine used in oriental medicine. For example, Korean Patent Publication No. 2007-0070307 proposes a composition for improving sleep surface comprising various herbal ingredients in an antihistamine drug. Korean Patent No. 1020245 discloses a composition for improving the water surface comprising a licorice ethanol extract and / or a waxy ethanol extract.
하지만, 수면 장애를 치료하고/치료하거나, 수면 시간 증진 및/또는 수면 질을 개선, 향상시키는 동시에, 인체에 무해하여 안전성이 확보될 수 있는 천연 소재를 개발할 필요성은 여전히 남아 있다. However, there remains a need to develop natural materials that can treat and / or treat sleeping disorders, improve sleeping time and / or sleep quality, and at the same time, be safe and harmless to the human body.
본 발명의 목적은 생체에 독성이 없으며 안전성이 확보될 수 있는 식물 추출물을 유효 성분으로 함유하는 수면 장애를 치료하기 위한 약학 조성물 및/또는 의약품을 제공하고자 하는 것이다. It is an object of the present invention to provide a pharmaceutical composition and / or a medicament for treating a sleep disorder containing an extract of a plant which is free from toxicity to a living body and can secure safety.
본 발명의 다른 목적은 식물 추출물을 유효 성분으로 함유하는 수면 시간 증진 및/또는 수면 질 개선용의 기능성 식품 조성물 및/또는 건강기능식품을 제공하고자 하는 것이다. Another object of the present invention is to provide a functional food composition and / or a health functional food for improving sleeping time and / or sleep quality, which comprises a plant extract as an active ingredient.
전술한 목적을 가지는 본 발명의 일 측면에 따르면, 본 발명은 둥굴레(Polygonatum sp.) 추출물을 유효 성분으로 함유하는 수면 장애를 치료하기 위한 약학 조성물을 제공한다.According to an aspect of the present invention, there is provided a pharmaceutical composition for treating a sleep disorder containing Polygonatum sp. Extract as an active ingredient.
일례로, 상기 둥굴레 추출물은 둥굴레 열수 추출물, 둥굴레 유기용매 추출물, 둥굴레 효소 분해 추출물 중에서 선택되는 적어도 1종의 추출물인 것을 특징으로 한다. For example, the dongguloe extract is characterized by being at least one kind of extract selected from dongguljang hot water extract, dongguloe organic solvent extract, and dongguljie enzymatic decomposition extract.
예시적인 실시형태에서, 상기 둥굴레 추출물은 층층갈고리둥굴레(Polygonatum sibiricum) 추출물, 대잎둥굴레(Polygonatum falcatum) 추출물 및 둥굴레(Polygonatum odoratum)로 구성되는 군에서 선택되는 적어도 1종의 추출물을 포함할 수 있다. In an exemplary embodiment, the Drosophila extract is selected from the group consisting of Polygonatum sibiricum extract, Polygonatum falcatum extract, and polygonatum odoratum . The extract may be at least one selected from the group consisting of falcatum extract and polygonatum odoratum .
예를 들어, 상기 둥굴레 효소 분해 추출물은 상기 둥굴레를 셀룰라아제, 비스코자임 및 베타-글루카나아제로 구성되는 군에서 선택되는 적어도 1종의 효소로 처리하여 얻어질 수 있다. For example, the Drosophila hydrolyzate can be obtained by treating the drosophila with at least one enzyme selected from the group consisting of cellulase, viscose, and beta-glucanase.
상기 둥굴레 추출물은 상기 약학 조성물 중에 0.01 ~ 1000 ㎎/㎖의 농도로 함유될 수 있다. The extract may be contained in the pharmaceutical composition at a concentration of 0.01 to 1000 mg / ml.
또한, 본 발명의 다른 측면에 따르면, 본 발명은 둥굴레(Polygonatum sp.) 추출물을 유효 성분으로 함유하는 수면 시간 증진 및 수면 질 개선용 기능성 식품 조성물을 제공한다. According to another aspect of the present invention, there is provided a functional food composition for improving sleeping time and improving sleep quality, which comprises Polygonatum sp. Extract as an active ingredient.
본 발명에 따르면, 둥굴레 추출물을 유효 성분으로 함유하는 수면 장애를 치료하기 위한 약학 조성물 및 수면 시간 증진 및/또는 수면 질 개선용의 기능성 식품 조성물을 제안한다. According to the present invention, there is provided a pharmaceutical composition for treating a sleep disorder containing an extract of Aspergillus oryzae as an active ingredient, and a functional food composition for improving sleeping time and / or sleep quality.
특히, 본 발명에 따른 둥굴레 추출물을 동물에 투여하였을 경우, 수면 잠복기(sleep latency)가 감소하고 수면 시간(sleep duration)이 증가하였으며, 이 둥굴레 추출물은 생체 내 신경 신호체계의 하나인 GABA A 수용체의 벤조다이아제핀 결합 부위에 결합할 수 있다. In particular, when the extract of Dangguul extract according to the present invention was administered to animals, the sleep latency decreased and the sleep duration increased, and the Dangguul extract was administered to the animals in the presence of GABA A receptor Benzodiazepine binding site.
본 발명에 따른 조성물 중의 유효 성분으로 함유되는 둥굴레 추출물은 식용으로 섭취 가능한 천연 소재에서 유래한 것으로, 생체에 대한 독성이나 부작용이 없을 것으로 예상된다. 따라서 인체에 대한 안전성이 확보될 수 있는 의약품 또는 식품 중의 유효 활성 성분으로 적용될 수 있으며, 의약 산업 및 식품 산업에서 활용될 수 있다. It is expected that the extract of Dandelion as an active ingredient in the composition according to the present invention is derived from a natural material which can be ingested for food and has no toxicity or side effects to the living body. Therefore, it can be applied as an active ingredient in pharmaceuticals or foods which can secure human safety, and can be utilized in the pharmaceutical industry and the food industry.
또한, 국내에서 쉽게 구입할 수 있는 둥굴레를 일종의 유전자원으로 활용함으로써, 나고야 의정서의 발효와 관련하여 국내의 유전자원을 확보할 수 있을 것으로 기대된다. In addition, it is expected that it will be possible to secure the domestic genetic resources in connection with the entry into force of the Nagoya Protocol by using the donggulle, which can be purchased easily in Korea, as a genetic resource.
도 1과 도 2는 각각 본 발명의 예시적인 실시예에 따라 둥굴레 추출물을 투여한 래트(rat)를 대상으로 수면 시간 증진과 관련한 뇌전도(EEG; electroencephalogram)을 측정한 방법을 개략적으로 나타낸 모식도이다.
도 3은 본 발명의 예시적인 실시예에 따라 둥굴레 추출물 및 효소 분해물의 수면 증진 효능 분석 결과를 나타낸 그래프이다. CON은 음성 대조군이고, PSE는 층층갈고리둥굴레 열수 추출물, 효소 B는 층층갈고리둥굴레 베타-글루카나아제 효소 분해물, 효소 C는 층층갈고리둥굴레 셀룰라아제 효소 분해물, 효소 V는 층층갈고리둥굴레 비스코자임 효소 분해물을 래트에 투여한 것을 나타낸다. 음성 컨트롤에 대한 유의미한 수준을 별표로 표시하였다.
도 4는 본 발명의 예시적인 실시예에 따라 다양한 둥굴레 속 추출물의 수면 증진 효능 분석 결과를 나타낸 그래프이다. CON은 음성 대조군이고, 1은 층층갈고리둥굴레 추출물, 2는 대잎둥굴레, 3번과 4번은 둥굴레(Polygonatum odoratum)로 잠정 추정되는 둥굴레(Polygonatum sp.) 추출물을 래트에 투여한 것을 나타낸다. 음성 컨트롤에 대한 유의미한 수준을 별표로 표시하였다.
도 5는 본 발명의 예시적인 실시예에 따라 둥굴레 물 추출물의 GABA A 벤조다이아조펜 수용체에 대한 결합 능력을 측정한 결과를 나타낸 그래프이다. FIG. 1 and FIG. 2 are schematic views schematically illustrating a method of measuring an electroencephalogram (EEG) related to sleeping time enhancement in a rat administered with a Dandelion extract according to an exemplary embodiment of the present invention.
FIG. 3 is a graph showing the results of analyzing the sleep enhancement efficacy of the extracts of Dinggoli and enzyme hydrolysates according to an exemplary embodiment of the present invention. CON is a negative control, PSE is a stratified egg gangrene hydrothermal extract, Enzyme B is a stratum gingival sphagnum Beta-glucanase hydrolyzate, Enzyme C is a stratified sphagnum gangrene cellulase hydrolyzate, ≪ / RTI > Significant levels of voice control were marked with asterisks.
FIG. 4 is a graph showing the results of analyzing the sleep enhancement efficacy of various extracts of Aspergillus oryzae according to an exemplary embodiment of the present invention. CON represents a negative control, 1 represents a layered gull, 2 a polygonatum, 3 and 4 polygonatum odoratum , and a Polygonatum sp. Extract. Significant levels of voice control were marked with asterisks.
FIG. 5 is a graph showing the results of measuring the binding capacity of GABA A benzodiazepine receptors to water extracts of Dangguljoe according to an exemplary embodiment of the present invention.
이하, 필요한 경우에 첨부하는 도면을 참조하면서 본 발명을 보다 상세하게 설명한다. Hereinafter, the present invention will be described in more detail with reference to the accompanying drawings where necessary.
[둥굴레 추출물 및 그 제조 방법][Danglingu extract and its production method]
본 발명에 따라 수면 장애를 치료하기 위한 약학 조성물 및/또는 수면 시간 증진 및/또는 수면 질 개선을 위한 기능성 식품 조성물 중에 유효 성분으로 둥굴레(Polygonatum sp.) 추출물이 사용될 수 있다. According to the present invention, Polygonatum sp. Extract may be used as an active ingredient in a pharmaceutical composition for treating a sleeping disorder and / or a functional food composition for improving sleeping time and / or sleep quality.
본 발명에서 유효 성분으로 사용된 둥굴레는 백합과에 속하는 다년생 초본으로 북미, 유럽, 동북아시아 등 전세계적으로 40 여종이 분포하며, 국내에는 16 여종이 분포하는 것으로 알려져 있다. 하나의 예시적인 실시형태에서, 유효 성분으로 사용될 수 있는 둥굴레 추출물은 층층갈고리둥굴레(Polygonatum sibiricum) 추출물, 대잎둥굴레(Polygonatum falcatum) 추출물 및 둥굴레(Polygonatum odoratum)로 구성되는 군에서 선택되는 적어도 1종의 추출물이다. 특히 바람직하게는 층층갈고리둥굴레 추출물이다. It is a perennial herb that belongs to the Lily family. It is known that about 40 species are distributed throughout North America, Europe, Northeast Asia, and 16 species are distributed in Korea. In one exemplary embodiment, the Drosophila Extract, which can be used as an active ingredient, is a polygonatum sibiricum extract, Polygonatum falcatum extract, and polygonatum odoratum . Particularly preferably, the stratum corneum is an extract of Sambrook.
예를 들어, 둥굴레 추출물은 열수 추출물과 같은 물 추출물, 유기 용매 추출물은 물론이고, 이들 용매로 추출한 조-추출물을 적절한 용매로 분획하여 얻어진 추출물, 효소 분해 추출물 및 이들의 혼합물을 적절한 추출 용매로 추출하여 얻어질 수 있다. 하나의 예시적인 실시형태에서, 본 발명에 따른 둥굴레 추출물을 제조하기 위하여, 세척한 둥굴레의 뿌리줄기(근경)를 적절한 크기, 예를 들어 1 ~ 5 ㎝로 절단하고 용매 추출 및/또는 효소 처리를 수행할 수 있다. For example, the extracts of Dangwolla extract are not only water extracts such as hot water extracts, organic solvent extracts, extracts obtained by fractionating the crude extracts extracted with these solvents with appropriate solvents, enzyme decomposition extracts and mixtures thereof, . In one exemplary embodiment, in order to produce the extract of Dangongleae according to the present invention, the roots of the washed Dandelion are cut to an appropriate size, for example, 1 to 5 cm, and subjected to solvent extraction and / or enzyme treatment Can be performed.
둥굴레 추출물을 제조하기 위하여 사용되는 추출 용매는 열수와 같은 물은 물론이고, 유기 용매가 사용될 수 있다. 추출 용매로 사용될 수 있는 유기 용매는 특별히 제한되지는 않지만, 40% 내지 100%의 메탄올, 에탄올 등의 탄소수 1 내지 6의 저급 알코올은 물론이고, 식물 추출물을 얻기 위하여 통상적으로 사용되는 다른 유기 용매가 사용될 수 있다. As the extraction solvent used for preparing the extract, it is possible to use an organic solvent as well as water such as hot water. The organic solvent which can be used as the extraction solvent is not particularly limited, but may be a low alcohol having 1 to 6 carbon atoms such as methanol and ethanol at 40% to 100%, and other organic solvents usually used for obtaining plant extracts Can be used.
저급 알코올은 탄소수 1 내지 6의 알코올일 수 있다. 예를 들어, 저급 알코올로는 메탄올, 에탄올, 프로판올, 부탄올, 노말-프로판올, 이소-프로판올, 노말-부탄올, 1-펜탄올, 2-부톡시에탄올 또는 에틸렌글리콜 등을 이용할 수 있다. 유기 용매는 이 외에도 아세트산, DMFO(dimethyl-formamide), DMSO(dimethyl sulfoxide) 등의 극성 용매, 아세토나이트릴, 에틸 아세테이트, 메틸 아세테이트, 플루오로알칸, 펜탄, 2,2,4-트리메틸펜탄, 데칸, 사이클로헥산, 사이클로펜탄, 디이소부틸렌, 1-펜텐, 1-클로로부탄, 1-클로로펜탄, o-자일렌, 디이소프로필 에테르, 2-클로로프로판, 톨루엔, 1-클로로프로판, 클로로벤젠, 벤젠, 디에틸 에테르, 디에틸 설파이드, 클로로포름, 디클로로메탄, 1,2-디클로로에탄, 아닐린, 디에틸아민, 에테르, 사염화탄소 및 THF(Tetrahydrofuran) 등의 비극성 용매를 사용할 수도 있다.The lower alcohol may be an alcohol having 1 to 6 carbon atoms. For example, as the lower alcohol, methanol, ethanol, propanol, butanol, n-propanol, iso-propanol, n-butanol, 1-pentanol, 2-butoxyethanol or ethylene glycol can be used. The organic solvent may be a polar solvent such as acetic acid, dimethyl-formamide (DMFO) or dimethyl sulfoxide (DMSO), acetonitrile, ethyl acetate, methyl acetate, fluoroalkane, pentane, 2,2,4-trimethylpentane, decane 1-pentene, 1-chlorobutane, 1-chloropentane, o-xylene, diisopropyl ether, 2-chloropropane, toluene, 1- chloropropane, chlorobenzene , Non-polar solvents such as benzene, diethyl ether, diethyl sulfide, chloroform, dichloromethane, 1,2-dichloroethane, aniline, diethylamine, ether, carbon tetrachloride and THF (tetrahydrofuran) may be used.
예시적인 실시형태에서, 추출 유기 용매는 바람직하게는 40% 내지 100%의 메탄올, 에탄올 등의 탄소수 1 내지 6의 알코올, 더욱 바람직하게는 40% 내지 80% 에탄올일 수 있다. 상기 추출물, 예를 들어 40 내지 100℃의 열수 추출물 및/또는 바람직하게는 40% 내지 100%의 메탄올, 에탄올 등의 탄소수 1 내지 6의 알코올 추출물, 더욱 바람직하게는 에탄올 추출물 및/또는 효소 분해 추출물은 래트 모델에 대하여 수면 시간과 숙면 시간을 증가시키는 것으로 확인되었다. 본 발명의 둥굴레 추출물은 일반적으로 추출 기기, 초음파분쇄 추출기 또는 분획기를 이용할 수 있다. In an exemplary embodiment, the extraction organic solvent may preferably be 40% to 100% methanol, an alcohol having 1 to 6 carbon atoms such as ethanol, more preferably 40% to 80% ethanol. The above extract, for example, a hot water extract at 40 to 100 ° C and / or preferably an alcohol extract having 1 to 6 carbon atoms such as 40% to 100% of methanol or ethanol, more preferably an ethanol extract and / Was found to increase sleep time and sleep time for the rat model. In general, the extract of Dangguire of the present invention may be an extraction device, an ultrasonic pulverizing extractor or a fractionator.
예를 들어, 열수 추출물을 얻기 위하여 건조된 둥굴레를 적절한 크기로 분쇄 또는 절단하여 추출 용기에 넣고, 대략 10 ~ 30 배 가량의 물을 가하고 40 ~ 100℃에서 가열하여 2 ~ 12시간 추출액을 얻을 수 있다. 또한, 에탄올 추출물을 얻기 위하여, 건조된 둥굴레를 적절한 크기로 절단하여 40% ~ 100%, 바람직하게는 40% ~ 80%의 에탄올을 50 ~ 70℃에서 2 ~ 10시간 환류 추출하여 추출액을 얻을 수 있다. 필요한 경우, 둥굴레의 추출 효율을 높이기 위하여 상기 과정을 수회 이상 반복 수행할 수 있다. 상기 추출액을 여과하고, 감압농축하여 분말 형태의 둥굴레 추출물을 제조할 수 있다. 필요한 경우, 사용할 때까지 급속 냉동고(deep freezer)에 보관하거나 동결 건조할 수 있으며, 분말을 증류수 또는 통상의 용매에 녹여 사용할 수도 있다. For example, in order to obtain a hot-water extract, the dried siberia is pulverized or cut into an appropriate size, placed in an extraction container, added with about 10 to 30 times of water and heated at 40 to 100 ° C to obtain an extract for 2 to 12 hours have. In order to obtain an ethanol extract, the dried siberuli was cut to an appropriate size, and 40 to 100%, preferably 40 to 80% of ethanol was reflux-extracted at 50 to 70 ° C for 2 to 10 hours to obtain an extract have. If necessary, the above process may be repeated several times or more in order to increase the extraction efficiency of sodgola. The extract is filtered and concentrated under reduced pressure to give a powdery Danguryu extract. If necessary, it may be stored in a deep freezer or lyophilized until used, and the powder may be dissolved in distilled water or a conventional solvent.
필요한 경우, 용매로 추출한 추출물은 이후, 헥산, 메틸렌클로라이드, 아세톤, 에틸아세테이트, 에틸에테르, 클로로포름, 물 및 이들의 혼합물로 이루어진 군으로부터 선택된 용매로 분획과정을 더욱 실시할 수 있다. 바람직하게는 상기 분획용매는 에탄올 수용액, 물, 에틸아세테이트, 클로로포름 또는 클로로포름과 에탄올 혼합액일 수 있다. 아울러, 제조된 추출물 또는 상기 분획 과정을 수행하여 수득한 분획물은 이후 여과하거나 농축 또는 건조과정을 수행하여 용매를 제거할 수 있으며, 여과, 농축 및 건조를 모두 수행할 수 있다. 구체적으로 상기 여과는 여과지를 이용하거나 감압여과기를 이용할 수 있으며, 상기 농축은 감압 농축기, 일례로 회전 증발기를 이용하여 감압농축할 수 있으며, 상기 건조는 일례로 동결건조법으로 수행할 수 있다. If necessary, the solvent-extracted extract can then be further fractionated with a solvent selected from the group consisting of hexane, methylene chloride, acetone, ethyl acetate, ethyl ether, chloroform, water and mixtures thereof. Preferably, the fraction solvent may be an aqueous solution of ethanol, water, ethyl acetate, chloroform, or a mixture of chloroform and ethanol. In addition, the produced extract or the fraction obtained by performing the fractionation process may be filtered, concentrated or dried to remove the solvent, and may be subjected to both filtration, concentration, and drying. Specifically, the filtration may be performed using a filter paper or a vacuum filter. The concentration may be performed by a reduced pressure concentrator, for example, a rotary evaporator, and the drying may be performed by, for example, a freeze drying method.
둥굴레 효소 분해 추출물은 셀룰라아제, 비스코자임 및 베타-글루카나아제로 이루어진 적어도 1종의 분해 효소 및/또는 효소 칵테일을 첨가한 물로 40 내지 60에서 2 - 12시간 둥굴레를 추출하여 제조할 수 있다. 열수 추출, 유기 용매 추출 및 효소 분해 추출과 관련해서 상기에 기재된 실시형태는 모두 예시일 뿐으로, 본 발명이 이에 한정되지 않는다. The extract of Dandelion's enzyme may be prepared by extracting Dandelion at 40 to 60 for 2 to 12 hours with water containing at least one of a degrading enzyme consisting of a cellulase, a viscose and a beta-glucanase and / or an enzyme cocktail. All of the above-described embodiments relating to hot water extraction, organic solvent extraction and enzymatic decomposition extraction are merely examples, and the present invention is not limited thereto.
필요하다면, 둥굴레의 열수 추출, 유기 용매 추출, 효소 분해 추출 및 분획물을 물이나 유기 용매에 현탁시킨 후, 적절한 유출 용매(예를 들어 헥산/ 에틸아세테이트, 에틸아세테이트, 메탄올 등)을 사용하여 칼럼 크로마토그래피를 수행하여 보다 정제된 형태의 둥굴레 분획물을 얻을 수도 있다. 대안적인 실시형태에서, 상기 추출물이나 분획물을 적절한 분자량의 컷-오프 값을 가지는 한외 여과공정을 통과시켜 얻은 분획일 수도 있다. If necessary, the extracts of hydrothermal extract, organic solvent extraction, enzyme digestion and fractions of Dandelion are suspended in water or an organic solvent, and then purified by column chromatography using a suitable elution solvent (for example, hexane / ethyl acetate, ethyl acetate, It is also possible to obtain a more purified form of the Aspergillus sp. In an alternative embodiment, the extract or fraction may be a fraction obtained by passing through an ultrafiltration process having a cut-off value of appropriate molecular weight.
따라서, 본 명세서에서 '둥굴레 추출물'은 전술한 바와 같이, 추출, 분획 또는 정제의 각 단계에서 얻어지는 모든 추출액, 분획 및 정제물, 그들의 희석액, 농축액 또는 건조물을 모두 포함하는 개념이다.Accordingly, in the present specification, the term "dongguloe extract" is a concept including all the extracts, fractions and tablets obtained in each step of extraction, fractionation or purification, diluted solutions thereof, concentrates or dried products as described above.
수면의 종류에는 크게 REM (Rapid Eye Movement) 수면과 Non-REM (Non-rapid eyes movements) 수면이 존재한다. REM 수면은 수면 시에 베타-파가 방출되는 상태로 수면 시에 몸을 뒤척이며, 아침이 될수록 많아진다. 반면 Non-REM 수면은 델타-파가 방출되는 상태로 깊은 수면(숙면)에 해당한다. 인간이 수면을 취할 때, 일반적으로 Non-REM 수면을 3,4회 정도 겪는데, 횟수나 시간이 부족하게 되면, 수면 부족을 느끼게 된다. 따라서 수면의 질에 있어서 REM 수면 시간을 연장시키는 것보다는 Non-REM 수면 시간을 연장시키는 것이 중요하다. 본 발명의 예시적인 실시예에 따르면, 둥굴레 추출물을 동물 모델에 투여하고 뇌전도 분석을 수행한 결과, 대조군에 비하여 총 수면 시간과 숙면 시간(Non-REM 시간)을 크게 증가시킨다(도 3 내지 도 4 참조). Types of sleep include Rapid Eye Movement (REM) sleep and Non-rapid eyes movements (REM) sleep. REM sleep is a state in which beta-waves are released at the time of sleep, and the body turns around at the time of sleep and becomes more and more in the morning. Non-REM sleep, on the other hand, corresponds to deep sleep (sleep) with delta-waves being emitted. When humans take sleep, they generally experience non-REM sleep three or four times, and when the number or time is short, they feel sleep deprivation. Therefore, it is important to extend Non-REM sleep time rather than prolong REM sleep time in the quality of sleep. According to an exemplary embodiment of the present invention, when the extract of Dandelion is administered to an animal model and electroencephalogram analysis is performed, the total sleep time and the sleeping time (Non-REM time) are greatly increased as compared with the control group Reference).
특히, 둥굴레 추출물은 감마-아미노부티르산(gamma-aminobutyric acid; GABA)의 A 수용체(GABAA 수용체) 중의 벤조다이아제핀 결합 부위에 결합함으로써, 수면 잠복기(sleep latency)를 감소시키고, 수면 시간(sleep duration)을 증가시킨다(도 5 참조). Particularly, the extract of Dandelion extract binds to the benzodiazepine binding site in the A receptor (GABA A receptor) of gamma-aminobutyric acid (GABA), thereby reducing sleep latency and sleep duration ) (See Fig. 5).
GABAA 수용체는 신경신호전달과 관련된 수용체로써 수면 유도 및 개선 효과와 관련되어 있으며 이 외에 불안 완화, 경련 개선, 진정 작용과 관련된 수용체로 연구되었다. 현재 GABAA 수용체에 작용하는 약물이 연구되어 판매되고 있으며, 그 종류는 벤조다이아제핀(benzodiazepine), 디아제팜(diazepam) 등이 있고, 길항제로서는 플루마제닐(flumazenil)이 존재한다. 현재 판매되는 의약품의 경우 수면증진에 효과가 있는 것으로 알려져 있으나 의존성이 있으며 부작용을 초래하는 결과가 보고되어 지속적으로 섭취 또는 복용하는데 한계가 있다. 반면, 본 발명에 따라 유효 성분으로 사용되는 둥굴레 추출물은 천연 소재에서 유래한 것으로 생체에 대한 독성이나 부작용이 없을 것으로 예상되므로 안전성이 확보될 수 있다. GABA A receptors are receptors involved in neurotransmission and are associated with sleep induction and improvement, as well as receptors associated with anxiety relief, seizure improvement, and sedation. Current GABA A Drugs that act on receptors have been studied and marketed, including benzodiazepine, diazepam, and flumazenil as an antagonist. Drugs currently on the market are known to be effective in improving sleep, but they are dependent and there are reports of side effects that limit the ability to continue taking or taking. On the other hand, according to the present invention, the extract of Dandelion used as an active ingredient is expected to have no toxicity or adverse effects on the living body, which is derived from natural materials, so that safety can be secured.
하나의 예시적인 실시형태에서, 본 발명에 따른 유효 성분인 둥굴레 추출물은 0.01 ~ 1000 ㎎/㎖, 바람직하게는 0.1 ~ 1000 ㎎/㎖, 더욱 바람직하게는 1 ~ 500 ㎎/㎖의 농도로 포함될 수 있지만, 본 발명이 이에 한정되지는 않는다. In one exemplary embodiment, the extract of Dandelion as an active ingredient according to the present invention may be contained at a concentration of 0.01 to 1000 mg / ml, preferably 0.1 to 1000 mg / ml, more preferably 1 to 500 mg / ml. However, the present invention is not limited thereto.
[약학 및 식품공학적 응용][Pharmacy and Food Engineering Application]
전술한 바와 같이, 본 발명에 따른 둥굴레 추출물은 수면 시간 및 NON-REM 수면으로 대표될 수 있는 숙면 시간을 향상시킨다. 따라서, 본 발명의 일 측면에 따르면, 본 발명은 둥굴레 추출물을 유효 성분으로 함유하는 수면 장애(sleep disturbance 및/또는 sleep disorder), 예를 들어 불면증(insomnia)를 치료하기 위한 약학 조성물 및/또는 의약품에 관한 것이다. As described above, the extract of Dunguryul according to the present invention improves the sleeping time which can be represented by the sleeping time and the NON-REM sleeping. Therefore, according to one aspect of the present invention, the present invention provides a pharmaceutical composition and / or a pharmaceutical composition for treating sleep disturbance and / or sleep disorder, for example insomnia, .
본 발명의 둥굴레 추출물의 약학적 투여 형태는 이들의 약학적 허용가능한 염의 형태로도 사용될 수 있고, 또한 단독으로 또는 타 약학적 활성 화합물과 결합뿐만 아니라 적당한 집합으로 사용될 수 있다. 즉, 수면 장애를 치료하기 위한 약학 조성물은 둥굴레 추출물을 유효성분으로 단독으로 포함할 수 있고, 이외 제형, 사용방법 및 사용목적에 따라 추가성분 즉, 약학적으로 허용되거나 영양학적으로 허용되는 담체, 부형제, 희석제 또는 부성분을 추가로 포함할 수 있다. The pharmacological dosage forms of the extracts of the present invention can be used in the form of their pharmaceutically acceptable salts, and they can be used alone or in combination with other pharmacologically active compounds as well as in suitable aggregates. That is, the pharmaceutical composition for treating a sleep disorder can contain Dangguul extract as an active ingredient alone, and may further contain additional components such as a pharmaceutically acceptable or nutritionally acceptable carrier, An excipient, a diluent or a subcomponent.
일례로, 본 발명에 따른 둥굴레 추출물을 포함하는 약학 조성물은, 각각 통상의 방법에 따라 산제, 과립제, 정제, 캡슐제, 현탁액, 에멀젼, 시럽, 에어로졸 등의 경구형 제형, 외용제, 좌제 및 멸균 주사용액의 형태로 제형화하여 사용될 수 있다. 또한, 본 발명의 수면 장애 치료용 약학 조성물의 제형은 사용방법에 따라 바람직한 형태일 수 있으며, 특히 포유동물에 투여된 후 활성 성분의 신속, 지속 또는 지연된 방출을 제공할 수 있도록 당업계에 공지된 방법을 채택하여 제형화할 수 있다. 구체적인 제형의 예로는 경고제, 과립제, 로션제, 리니멘트제, 리모나데제, 산제, 시럽제, 안연고제, 액제, 에어로솔제, 엑스제(EXTRACTS), 엘릭실제, 연고제, 유동엑스제, 유제, 현탁제, 전제, 침제, 점안제, 정제, 좌제, 주사제, 주정제, 캅셀제, 크림제, 환제, 연질 또는 경질 젤라틴 캅셀 등이 있다. For example, the pharmaceutical composition comprising the extract of Dangongleae according to the present invention may be administered orally or parenterally in the form of powders, granules, tablets, capsules, suspensions, emulsions, oral preparations such as syrups and aerosols, external preparations, Can be formulated in the form of a solution. In addition, the formulation of the pharmaceutical composition for the treatment of sleep disorders of the present invention may be in a desirable form depending on the method of use, and may be in a form suitable for administration to mammals such as those known in the art to provide rapid, sustained or delayed release of the active ingredient Method can be adopted and formulated. Exemplary formulations include, but are not limited to, excipients, granules, lotions, liniments, rimonadense, powders, syrups, ointments, liquids, aerosols, EXTRACTS, elixirs, ointments, Suspensions, premixes, infusions, eye drops, tablets, suppositories, injections, main tablets, capsules, creams, pills, soft or hard gelatin capsules.
예를 들어, 본 발명의 둥굴레 추출물은 임상 투여 시에 경구 또는 비경구의 여러 가지 제형으로 투여될 수 있는데, 제제화할 경우에는 보통 사용하는 충진제, 증량제, 결합제, 습윤제, 계면활성제, 항응집제, 윤활제, 습윤제, 향료, 유화제 또는 방부제와 같은 희석제 또는 부형제 등을 더욱 포함할 수 있으며, 경구 또는 비경구 모두 사용할 수 있다. For example, the extract of the present invention may be administered orally or parenterally in various dosage forms during clinical administration. In the case of formulation, the fillers, extenders, binders, wetting agents, surfactants, anticoagulants, lubricants, A wetting agent, a flavoring agent, an emulsifying agent or an antiseptic agent, and the like, and either orally or parenterally may be used.
예를 들어, 둥굴레 추출물을 포함하는 약학 조성물에 포함될 수 있는 담체, 부형제 및 희석제로는 락토오스, 덱스트로스, 수크로스, 솔비톨, 만니톨, 자일리톨, 에리스리톨, 말티톨, 전분, 아카시아 고무, 알지네이트, 젤라틴, 칼슘 포스페이트, 칼슘 실리케이트, 셀룰로스, 메틸 셀룰로스, 미정질 셀룰로스, 폴리비닐 피롤리돈, 물, 메틸히드록시벤조에이트, 프로필히드록시벤조에이트, 탈크, 마그네슘 스테아레이트 및 광물유, 덱스트린, 칼슘카보네이드, 프로필렌글리콜, 리퀴드 파라핀, 생리식염수로 이루어진 군에서 선택된 1이상 일 수 있으나, 이에 한정되는 것은 아니며 통상의 담체, 부형제 또는 희석제 모두 사용 가능하다. Examples of the carrier, excipient and diluent which can be contained in the pharmaceutical composition including the extract of Danguryu extract include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium But are not limited to, calcium carbonate, phosphate, calcium silicate, cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil, dextrin, calcium carbonate, , Liquid paraffin, and physiological saline. However, it is not limited thereto, and any conventional carrier, excipient or diluent may be used.
경구 투여를 위한 고형 제제에는 정제, 환제, 산제, 과립제, 캡슐제 등이 포함되며, 이러한 고형 제제는 본 발명의 둥굴레 추출액에 적어도 하나 이상의 부형제 예를 들면, 전분, 칼슘카보네이트(calcium carbonate), 수크로스(sucrose) 또는 락토오스(lactose), 젤라틴 등을 섞어 조제된다. 또한 단순한 부형제 이외에 마그네슘 스티레이트 탈크 같은 윤활제들도 사용된다. 경구 투여를 위한 액상 제제로는 현탁제, 내용액제, 유제 및 시럽제 등이 해당되는데 흔히 사용되는 단순 희석제인 물, 리퀴드 파라핀 이외에 여러 가지 부형제, 예를 들면 습윤제, 감미제, 방향제 및 보존제 등이 포함될 수 있다. The solid preparations for oral administration include tablets, pills, powders, granules, capsules and the like. These solid preparations can be prepared by adding to the dungbore extract of the present invention at least one or more excipients such as starch, calcium carbonate, Sucrose, lactose, gelatin and the like. In addition to simple excipients, lubricants such as magnesium stearate talc are also used. Liquid preparations for oral administration include suspensions, solutions, emulsions and syrups. Various excipients such as wetting agents, sweeteners, fragrances and preservatives may be included in addition to water and liquid paraffin, which are commonly used simple diluents. have.
비경구 투여를 위한 제제에는 멸균된 수용액, 비수성용제, 현탁제, 유제, 동결건조 제제, 좌제가 포함된다. 비수성용제와 현탁제로는 프로필렌글리콜(propylene glycol), 폴리에틸렌글리콜, 올리브 오일과 같은 식물성 기름, 에틸올레이트와 같은 주사 가능한 에스테르 등이 사용될 수 있다. 좌제의 기제로는 위텝솔(witepsol), 마크로골, 트윈(tween) 61, 카카오지, 라우린지, 글리세롤, 젤라틴 및 글리세로젤라틴 등이 사용될 수 있다. 본 발명의 약학적 조성물은 비경구 투여시 피하주사, 정맥 주사 또는 근육내 주사를 통할 수 있다. Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Examples of non-aqueous solvents and suspensions include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, injectable esters such as ethyl oleate, and the like. As a suppository base, witepsol, macrogol, tween 61, cacao paper, laurin, glycerol, gelatin and glycerogelatin can be used. The pharmaceutical composition of the present invention can be administered parenterally, subcutaneously, intravenously or intramuscularly.
비경구 투여를 위한 형태로는 치약, 구강세정제, 국소 투여제(크림, 연고, 드레싱 용액, 분무제, 기타 도포제 등) 등을 들 수 있다. 상기 국소 투여제의 제형의 일예로는, 유효 성분인 둥굴레 추출물을 천연 섬유 또는 합성 섬유로 만든 거즈 등의 담체에 함침시킨 것일 수 있다. Examples of the form for parenteral administration include toothpastes, mouthwashes, topical administration agents (creams, ointments, dressing solutions, sprays, and other coating agents). An example of the formulation of the topical administration agent may be that the active ingredient Dulguyrae extract is impregnated with a carrier such as gauze made of natural fiber or synthetic fiber.
아울러, 상기 수면 장애 치료용 약학 조성물은 상기 유효 성분 외에 추가로 영양제, 비타민, 전해질, 풍미제, 착색제, 중진제, 펙트산 및 그의 염, 알긴산 및 그의 염, 유기산, 보호성 콜로이드 증점제, pH 조절제, 안정화제, 방부제, 글리세린, 알코올, 탄산음료에 사용되는 탄산화제 등을 추가로 함유할 수 있다. In addition, the pharmaceutical composition for the treatment of sleep disorders may further contain, in addition to the active ingredient, a nutrient, a vitamin, an electrolyte, a flavoring agent, a colorant, a thickening agent, a pectic acid and a salt thereof, an alginic acid and a salt thereof, , A stabilizer, a preservative, a glycerin, an alcohol, a carbonating agent used in a carbonated drink, and the like.
본 발명의 둥굴레 추출물을 포함하는 수면 장애 치료용 약학 조성물의 바람직한 투여량은 환자의 상태 및 체중, 질병의 정도, 약물형태, 투여경로 및 기간에 따라 다르지만, 당업자에 의해 적절하게 선택될 수 있다. 그러나 바람직한 효과를 위해서, 본 발명의 둥굴레 추출물은 1일 0.01 내지 1000 ㎎/㎏, 바람직하게는 0.1 내지 1000 ㎎/㎏의 양으로 투여하는 것이 좋다. 투여는 하루에 한번 투여할 수도 있고, 수회 나누어 투여할 수도 있다. 상기 투여량은 어떠한 면으로든 본 발명의 범위를 한정하는 것은 아니다. The preferred dosage of the pharmaceutical composition for treating sleep disorders, which comprises the extract of the present invention, depends on the condition and the weight of the patient, the degree of the disease, the drug form, the administration route and the period, but can be appropriately selected by those skilled in the art. However, in order to obtain the desired effect, it is preferable to administer the extract of Dandelion seed of the present invention in an amount of 0.01 to 1000 mg / kg, preferably 0.1 to 1000 mg / kg per day. The administration may be carried out once a day or divided into several times. The dose is not intended to limit the scope of the invention in any way.
한편, 본 발명의 다른 측면에 따르면, 본 발명은 둥굴레 추출물을 유효 성분으로 함유하는 수면 시간 증진 및/또는 수면 질 개선을 위한 기능성 식품 조성물 및/또는 건강기능식품에 관한 것이다. 본 발명의 기능성 식품 조성물의 예는 식품, 식품첨가제, 음료 또는 음료 첨가제를 들 수 있다. According to another aspect of the present invention, there is provided a functional food composition and / or a health functional food for improving sleeping time and / or sleep quality, comprising the extract of Dandelion as an active ingredient. Examples of the functional food composition of the present invention include food, food additive, beverage or beverage additive.
예를 들어, 본 발명에 따른 둥굴레 추출물을 함유할 수 있는 식품은 각종 식품류, 음료, 껌, 캔디, 차, 비타민 복합제, 기능성 식품 등이 있으며 식품의 종류에는 특별한 제한이 없다. 구체적으로, 둥굴레 추출물을 첨가할 수 있는 식품의 예는 육류, 소시지, 빵, 초콜릿, 캔디류, 스넥류, 과자류, 피자, 라면, 기타 면류, 검류, 아이스크림류를 포함하는 낙농제품, 각종 스프, 음료수, 차, 드링크제, 알코올 음료 및 비타민 복합제 등이 있으며, 통상적인 의미에서의 건강식품을 모두 포함한다. For example, the food that can contain the extract of Dangguire according to the present invention includes various foods, beverages, gums, candies, tea, vitamin complex, and functional food. Specifically, examples of foods in which the extract can be added include meat, sausage, bread, chocolate, candy, snack, confectionery, pizza, ramen, other noodles, dairy products including ice cream, Tea, a drink, an alcoholic beverage, and a vitamin complex, all of which include health foods in a conventional sense.
추가로, 본 발명에서 식품에는 특수영양식품(예, 조제유류, 영,유아식 등), 식육 가공품, 어육제품, 두부류, 묵류, 면류(예, 라면류, 국수류 등), 건강보조식품, 조미식품(예, 간장, 된장, 고추장, 혼합장 등), 소스류, 과자류(예, 스넥류), 유가공품(예, 발효유, 치즈 등), 기타 가공식품, 김치, 절임식품(각종 김치류, 장아찌 등), 음료(예, 과실,채소류 음료, 두유류, 발효음료류, 아이스크림류 등), 천연조미료(예, 라면 스프 등), 비타민 복합제, 알코올 음료, 주류 및 그 밖의 건강보조식품류를 포함하나 이에 한정되지 않는다. 상기 식품, 음료 또는 식품첨가제는 통상의 제조방법으로 제조될 수 있다.In addition, in the present invention, the food may contain special nutritional foods (e.g., crude oil, spirits, infant food, etc.), meat products, fish products, tofu, jelly, noodles (Such as soy sauce, soybean paste, hot pepper paste, mixed sauce), sauces, confectionery (eg snacks), dairy products (eg fermented milk, cheese), other processed foods, kimchi, pickled foods But are not limited to, natural flavors (eg, ramen soup, etc.), vitamin complexes, alcoholic beverages, alcoholic beverages and other health supplement foods. The food, beverage or food additive may be prepared by a conventional production method.
기능성 식품에는 식품공학적으로 허용 가능한 식품 보조 첨가제를 포함할 수 있으며, 기능성 식품의 제조에 통상적으로 사용되는 적절한 담체, 부형제 및 희석제를 더욱 포함할 수 있다. 본 발명의 둥굴레 추출물을 식품 첨가물로 사용할 경우, 상기 둥굴레 추출물을 그대로 첨가하거나 다른 식품 또는 식품 성분과 함께 사용될 수 있고, 통상적인 방법에 따라 적절하게 사용될 수 있다. 상기 둥굴레 추출물을 유효성분으로 포함하는 기능성 식품 조성물은 그 제조에 통상적으로 사용하는 적절한 담체, 부형제 및 희석제를 더욱 포함할 수 있다. 유효 성분의 혼합양은 사용 목적(예방, 건강 또는 치료적 처치)에 따라 적합하게 결정될 수 있다. Functional foods may include food-grade acceptable food-aid additives and may further comprise suitable carriers, excipients and diluents conventionally used in the manufacture of functional foods. When the extract of the present invention is used as a food additive, the extract can be used as it is or can be used together with other food or food ingredients, and can be suitably used according to a conventional method. The functional food composition containing the extract as an active ingredient may further contain an appropriate carrier, excipient and diluent commonly used in the production thereof. The amount of the active ingredient to be mixed can be suitably determined according to the intended use (prevention, health or therapeutic treatment).
선택적으로 수면 시간 증진 및/또는 수면 질 개선을 위한 기능성 식품 조성물 중에 상기 둥굴레 추출물의 양은 전체 식품 중량의 0.01 중량% 내지 50 중량%로 포함될 수 있으며, 음료 조성물의 경우 식품 전체의 부피 100 ㎖을 기준으로 0.001 g 내지 50 g, 바람직하게는 0.01 g 내지 10 g의 비율로 포함될 수 있다. 그러나 건강 및 위생을 목적으로 하거나 또는 건강 조절을 목적으로 하는 장기간의 섭취의 경우에는 상기 양은 상기 범위 이하일 수 있으며, 안전성 면에서 아무런 문제가 없기 때문에 유효성분은 상기 범위 이상의 양으로도 사용될 수 있다. Alternatively, the functional food composition for improving the sleeping time and / or improving the sleep quality may comprise 0.01 to 50% by weight of the total food weight, and in the case of the beverage composition, the total volume of the food is 100 ml In a proportion of from 0.001 g to 50 g, preferably from 0.01 g to 10 g. However, in the case of long-term intake intended for health and hygiene purposes or for the purpose of controlling health, the amount may be less than the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount exceeding the above range.
기능성 식품 조성물은 정제, 캡슐제, 환제, 액제 등의 형태를 포함한다. 기능성 식품 조성물 또는 건강기능식품은 지시된 비율로 필수 성분으로서 둥굴레 추출물을 함유하는 외에는 다른 성분에는 특별한 제한이 없으며 통상의 음료와 같이 여러 가지 향미제 또는 천연 탄수화물과 같은 식품 보조 첨가제 성분을 추가 성분으로서 함유할 수 있다. The functional food composition includes forms such as tablets, capsules, pills, liquids and the like. The functional food composition or the health functional food is not particularly limited to the other ingredients except that it contains the extract as an essential ingredient in the indicated ratios and it is also possible to add a food supplementary ingredient such as various flavors or natural carbohydrates as an additional ingredient ≪ / RTI >
천연 탄수화물의 예는 포도당, 과당과 같은 단당류; 말토오스, 수크로오스 등과 같은 이당류; 덱스트린, 시클로덱스트린과 같은 다당류 등과 같은 통상적인 당은 물론이고, 자일리톨, 소르비톨, 에리트리톨 등의 당알코올이다. 상술한 것 이외의 향미제 또는 감미제로서 타우마틴, 스테비아 추출물(예를 들어 레바우디오시드 A, 글리시르히진)과 같은 천연 향미제/감미제; 사카린, 아스파르탐과 같은 합성 향미제/감미제 등을 사용할 수 있다. 상기 천연 탄수화물의 비율은 본 발명의 기능성 식품 조성물 100 ㎖당 일반적으로 약 1 내 지 20g, 바람직하게는 약 5 내지 12g이다. 상기 천연탄수화물의 비율은 본 발명에 따른 조성물 100 ㎖당 일반적으로 약 0.01~0.04 g, 바람직하게는 약 0.02~0.03 g 이다.Examples of natural carbohydrates include monosaccharides such as glucose and fructose; Disaccharides such as maltose, sucrose and the like; Sugar alcohols such as xylitol, sorbitol and erythritol, as well as conventional sugars such as polysaccharides such as dextrin and cyclodextrin. Natural flavors / sweeteners such as tautatin, stevia extract (e.g., rebaudioside A, glycyrrhizin) as flavorings or sweeteners other than those mentioned above; Synthetic flavorings / sweeteners such as saccharin and aspartame may be used. The ratio of the natural carbohydrate is generally about 1 to 20 g, preferably about 5 to 12 g per 100 mL of the functional food composition of the present invention. The ratio of the natural carbohydrate is generally about 0.01 to 0.04 g, preferably about 0.02 to 0.03 g per 100 ml of the composition according to the present invention.
상기 외에 본 발명의 기능성 식품 조성물은 여러 가지 영양제, 비타민, 광물(전해질), 합성 풍미제 및 천연 풍미제 등의 풍미제, 착색제 및 중진제(치즈, 초콜릿 등), 펙트산 및 그의 염, 알긴산 및 그의 염, 유기산, 보호성 콜로이드 증점제, pH 조절제, 안정화제, 방부제, 글리세린, 알코올 탄산음료에 사용되는 탄산화제 등을 함유할 수 있다. 그밖에 본 발명에 따른 기능성 식품 조성물 중에는 천연 과일 주스 및 과일 주스 음료 및 야채 음료의 제조를 위한 과육을 함유할 수 있다. 이러한 성분은 독립적으로 또는 조합하여 사용할 수 있다. 이러한 첨가제의 비율은 그렇게 중요하진 않지만 본 발명의 둥굴레 추출물 100 중량부 당 0 내지 약 20 중량부, 바람직하게는 0.01 ~ 1.0 중량부의 범위에서 선택되는 것이 일반적이다. In addition to the above, the functional food composition of the present invention may contain flavoring agents such as various nutrients, vitamins, minerals (electrolytes), synthetic flavors and natural flavors, colorants and heavies (cheese, chocolate etc.), pectic acid and its salts, And salts thereof, organic acids, protective colloid thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, carbonating agents used in alcoholic carbonated beverages, and the like. In addition, the functional food composition according to the present invention may contain natural fruit juice and pulp for the production of fruit juice drinks and vegetable drinks. These components may be used independently or in combination. Although the ratio of such additives is not so important, it is generally selected in the range of 0 to about 20 parts by weight, preferably 0.01 to 1.0 part by weight, per 100 parts by weight of the Dangongle extract of the present invention.
이하, 예시적인 실시예를 통하여 본 발명을 설명하지만, 본 발명이 하기 실시예에 기재된 발명으로 한정되지 않는다. Hereinafter, the present invention will be described with reference to exemplary embodiments, but the present invention is not limited to the invention described in the following embodiments.
실시예Example 1: One: 둥굴레Dingle 추출물 제조 Extract preparation
층층갈고리둥굴레(Polygonatum sibiricum) 추출물(PSE)을 다음과 같이 제조하였다. 층층갈고리둥굴레의 뿌리 줄기를 2.0 ㎝ 길이로 절단하여 건조시킨 둥굴레 건조물 200g과 물 3L를 5L 추출용기에 넣고 90℃에서 4시간 열수 추출 하였다. 추출물을 원심분리(8000 rpm, 20분)하여 얻어진 상층액을 감압농축하여 둥굴레 물 추출 농축액을 얻었다. 이를 분무 건조하여 둥굴레 물 추출 분말을 제조하였다. Layered hooks Polygonatum sibiricum ) extract (PSE) was prepared as follows. Layered hooks The roots of duckweed were cut into 2.0 ㎝ length and dried, and then 3 g of water and 3 g of water were put into a 5L extraction vessel and hot water was extracted at 90 ℃ for 4 hours. The extract was centrifuged (8000 rpm, 20 minutes), and the resulting supernatant was concentrated under reduced pressure to obtain a water extract concentrate of Sambrook. This was spray dried to prepare water extract powder of Dangguul.
실시예Example 2: 2: 둥굴레Dingle 추출물 제조 Extract preparation
층층갈고리둥굴레를 대신하여 대잎둥굴레(Polygonatum falcatum)를 사용한 것을 제외하고 실시예 1의 절차를 반복하여 둥굴레 추출물을 제조하였다. Instead of the stratified hooks, Polygonatum The procedure of Example 1 was repeated except that falcatum was used.
실시예Example 3~4: 3 to 4: 둥굴레Dingle 추출물 제조 Extract preparation
층층갈고리둥굴레를 대신하여 동정되지 않은 둥굴레(Polygonatum sp.)를 사용한 것을 제외하고 실시예 1의 절차를 반복하여 둥굴레 추출물을 제조하였다. The procedure of Example 1 was repeated except that Polygonatum sp., Which was not identified in place of the stratum corneum gill, was used to prepare the gill extract.
실시예Example 5: SD-rat을 이용한 뇌전도(electroencephalogram; EEG)을 통한 수면 시간 및 수면의 질 분석 5: Analysis of sleep time and sleep quality through electroencephalogram (EEG) using SD-rat
실시예 1에서 제조한 둥굴레 추출물을 동물 모델에 투여한 뒤에 뇌전도 테스트를 수행하였다(도 1 및 도 2 참조). 실험에 사용한 동물은 평균 체중 250 g의 Sprague-Dawley(SD 래트)를 대한바이오링크(DBL)로부터 제공받아 사용하였다. 실험 동물은 5일간 환경에 적응시킨 후, 임의로 각 처리군당 8마리씩 나누어 배치하였다. 실험동물 사육실 환경온도는 20~22, 상대습도는 50~55%로 유지하였고, 명암은 12시간 주기로 조절했으며 물과 사료는 자유 급식시켰다. 수술 시 isoflurane을 통한 흡입마취 후 정위고정기(stereotaxic device)에 고정하여 Paxinos 및 Watson 해부도에 따라 뇌전도(Electroencephalogram, EEG) 전극을 삽입하였다. An EEG extract prepared in Example 1 was administered to an animal model, and an electroencephalogram test was conducted (see FIGS. 1 and 2). Animals used in the experiments were Sprague-Dawley (SD rats) with an average weight of 250 g supplied from BioLink (DBL). Experimental animals were adapted to the environment for 5 days and then randomly assigned to 8 animals per treatment group. Experimental animals were maintained at ambient temperature of 20 ~ 22 and relative humidity of 50 ~ 55%. The darkness was controlled by 12 - hour cycle and water and feed were fed free. Surgery was performed by isoflurane anesthesia, fixed to a stereotaxic device, and an electroencephalogram (EEG) electrode was inserted according to the Paxinos and Watson anatomy.
수술이 끝난 동물은 일주일간 회복 시간을 거친 후 뇌전도 무선 송출기(EEG transmitter)를 부착시키고 시험 물질을 경구투여한 시점을 기준으로 오전 10시부터 오후 6시까지 15 mm/sec으로 뇌전도 측정하였다. 뇌전도 측정 후 기준기록(baseline recording), 대조기록(control recording), 실험기록(experimental recording)을 통한 뇌파의 전위 분석으로 총 수면시간(total sleep) 및 수면의 질(sleep quality)을 분석하였다. 수면의 종류에는 크게 REM (Rapid Eye Movement) 수면과 Non-REM (Non-rapid eyes movements) 수면이 존재한다. 수면의 질에 있어서 REM(rapid eyes movements) 수면시간 보다는 Non-REM (Non-rapid eyes movements) 수면시간의 연장이 중요하므로, 이들 시간을 분리하여 측정하였다. After the operation, the animals were allowed to recover for one week, and then the EEG transmitter was attached and the EEG was measured at 15 mm / sec from 10 am to 6 pm based on the oral administration of the test substance. Total sleep and sleep quality were analyzed by baseline recording, control recording, and experimental recording of electroencephalogram (EEG) analysis. Types of sleep include Rapid Eye Movement (REM) sleep and Non-rapid eyes movements (REM) sleep. Since the duration of non-rapid eye movements (REM) is more important than the rapid eye movements (REM) sleep time in the quality of sleep, these times were measured separately.
준비된 SD-rat에 각각 정상대조군(Control)은 소금물을 경구 투여하였고, 양성대조군으로 일반의약품인 길초근 추출물의 주성분인 valerenic acid 표준품을 80 ㎍/㎏ (길초근 추출물 160 ㎎/㎏ 상당)을 경구 투여하였고, 실험군으로 실시예 1에서 제조한 둥굴레 물 추출 분말(PSE) 160 ㎎/㎏을 경구 투여하였다. 이들의 EEG를 활용하여 10:00부터 18:00까지 총 8시간의 뇌파를 8일 동안 측정하였다. 측정된 뇌파를 총 수면시간, REM 수면시간과 Non-REM 수면시간으로 구분하여 분석하였다. Each of the prepared SD-rats was orally administered with salt water, and the positive control group was 80 ㎍ / kg (corresponding to 160 mg / kg of long root extract) of valerenic acid, which is a main component of the common herbal extract, And 160 mg / kg of water extract powder (PSE) prepared in Example 1 was orally administered to the experimental group. Using these EEGs, a total of 8 hours of EEG was measured from 10:00 to 18:00 for 8 days. The measured EEG was divided into total sleep time, REM sleep time and non-REM sleep time.
본 실시예에 따른 분석 결과를 표 1에 나타낸다. 양성대조군인 길초근 추출물 투여군에서 정상대조군(Control)에 비해 각성시간은 유의적으로 감소하고, 그에 따라 수면시간이 유의적으로 증가하였다. 수면시간 중 Non-REM 시간은 유의적으로 증가하고, REM 시간은 유의적으로 감소하여 수면시간의 증가는 Non-REM 시간 증가에 의한 것으로 나타났다. Table 1 shows the analysis results according to this embodiment. In the control group Gilchoung extract extract group, the awakening time was significantly decreased and the sleeping time was significantly increased compared to the control group. The non-REM time increased significantly and the REM time decreased significantly, indicating that the increase in sleep time was due to the increase in Non-REM time.
둥굴레 추출물을 투여하였을 때에는 정상대조군과 비교하여 각성시간의 유의적인 감소를 확인하였다. 그에 따라 둥굴레 추출물 투여군에서 수면시간이 유의적으로 증가하였으며, 정상대조군과 비교해 Non-REM 시간의 유의적인 증가와 REM 시간의 유의적인 감소를 확인하였다. 일반의약품으로 사용되는 길초근 추출물의 숙면 시간 (Non-REM)이 4.9 시간인 것에 비해 둥굴레 추출 분말의 숙면시간은 5.34 시간으로 약 19% 더 연장된 것을 볼 수 있었다. 따라서, 둥굴레 추출물은 일반의약품 보다 우수한 효능을 보임을 확인할 수 있었다.A significant decrease in awakening time was observed compared with the control group when Dulgore extract was administered. As a result, the sleeping time was significantly increased in the dongguloe extract group, and the significant increase of the non-REM time and the significant decrease of the REM time were confirmed as compared with the normal control group. The non-REM of Ganoderma lucidum extract used as a general medicine was 4.9 hours, while the sleep time of Dandongle extract powder was 5.34 hours, which was about 19% longer. Therefore, it was confirmed that Dangguire extract showed better efficacy than general medicine.
실시예Example 6: 6: 둥굴레Dingle 추출물의 추출 조건에 따른 수면 증진 기능 분석 Analysis of sleep enhancement function by extracting condition of extract
실시예 1에서 제조된 둥굴레 물 추출물의 추출 온도 및 시간에 따른 수면유도 효과를 비교하기 위해 다음과 같이 둥굴레 추출물을 제조한 뒤 실시예 5의 방법에 따라 뇌전도 테스트를 수행하여 뇌파를 분석하였다. 먼저 추출 온도에 따른 효과를 비교하기 위하여 추출온도를 각각 40~100℃로 하고 다른 조건은 실시예 1과 같이 추출하였다. 다른 추출 온도에 따라 제조된 둥굴레 추출물에 대한 뇌전도 분석 결과를 표 2에 나타낸다. In order to compare the water-induced induction effect of the water extract of Dangguljoe prepared in Example 1 according to the extraction temperature and time, the Dangguire extract was prepared as follows, and the electroencephalogram test was conducted according to the method of Example 5 to analyze the brain waves. In order to compare the effect according to the extraction temperature, the extraction temperature was set to 40 to 100 ° C, respectively, and other conditions were extracted as in Example 1. Table 2 shows the results of electroencephalogram analysis of the extracts of Aspergillus oryzae with different extracting temperatures.
둥굴레 열수 추출 온도가 80℃ 이상일 때, 정상대조군(CON)에 비해 수면시간 및 수면시간 중 Non-REM 시간은 유의적으로 증가하는 것을 확인하였다. The non-REM time of sleeping time and sleeping time was significantly higher than that of normal control (CON) when the temperature of hot water extraction was 80 ℃ or higher.
(℃)Extraction temperature
(° C)
한편, 추출 시간 별 수면 증진기능 확인을 위해 90℃에서 2 ~ 12시간 동안 원물 대비 가수량 15배를 사용하여 실시예 1과 같이 추출물을 제조하였다. 다른 추출 시간에 따라 제조된 둥굴레 추출물에 대한 뇌전도 분석 결과를 표 3에 나타낸다. 추출 시간이 2시간 이상일 때부터 정상대조군(CON)과 비교해 수면 시간 및 수면시간 중 Non-REM 시간이 유의적으로 증가하는 것을 확인할 수 있었으며, 추출 4시간 이후부터 수면 시간 및 수면시간 중 NREM 시간이 더 이상 증가하지 않는 것을 확인하였다. The extract was prepared as in Example 1 using 15 times the volume of water relative to the raw material at 90 ° C for 2 to 12 hours in order to confirm the water-increasing function by the extraction time. Table 3 shows the results of electroencephalogram analysis of the extracts of Dangwolle according to different extraction times. The non-REM time was significantly increased in the sleeping time and the sleeping time compared to the normal control (CON) from the extraction time of more than 2 hours. The NREM time of the sleeping time and the sleeping time after 4 hours of extraction And it was confirmed that it did not increase any more.
실시예Example 7: 7: 둥굴레Dingle 추출물의 효소 처리를 통한 수면 증진 기능 분석 Analysis of sleep enhancement function by enzymatic treatment of extract
실시예 1에서 사용한 층층갈고리둥굴레 줄기 뿌리 소재를 1시간 효소 분해를 진행한 뒤에 실시예 1의 절차를 반복하여 효소 분해된 둥굴레 추출물을 제조하였다. 층층갈고리둥굴레 뿌리 줄기 200 g에 15배 가수하고 효소 분해를 진행하였다. 효소 분해는 비스코자임(viscozyme; 제조사: Novozyme; 효소종류: arabanase, cellulase, β-glucanase, hemicellulase, xylanase, 최적온도: 55), 셀루클라스트(celluclast; 제조사: Novozyme; 효소종류: cellulase, 최적온도: 60), 베타-글루카나나아제(beta glucanase; Filterase, 제조사: DSM, 효소종류: beta-glucanase, 최적온도 60)를 이용하여 진행하였다. 효소 반응은 1시간 진행하였고 온도와 pH는 각각에 대한 최적 조건에서 진행하였다. 제조된 둥굴레 효소 분해물은 실시예 5의 절차를 반복하여 수면 효능을 평가하였다. 본 실시예에 대한 평가 결과를 도 3 및 표 4에 나타낸다. 효소 분해 추출물에서도 대조군에 비하여 총 수면 시간이 향상된 것을 확인하였다. The procedure of Example 1 was repeated after 1 hour of enzyme digestion of the layered hooks of the roots of Example 1, and the enzyme-digested Dangwolle extract was prepared. Layered hooks 200 g of Asiatic rootstock were subjected to enzyme digestion 15 times and hydrolyzed. Enzymatic degradation was carried out in the following manner: Viscozyme (manufacturer: Novozyme; enzyme type: arabanase, cellulase, β-glucanase, hemicellulase, xylanase, optimum temperature: 55), celluclast : 60), beta-glucanase (Filterase, manufacturer: DSM, enzyme type: beta-glucanase, optimum temperature 60). Enzyme reaction proceeded for 1 hour and temperature and pH proceeded under optimal conditions. The prepared sodglass enzyme degraded product was repeatedly subjected to the procedure of Example 5 to evaluate sleeping efficacy. The evaluation results of this embodiment are shown in Fig. 3 and Table 4. It was also found that the total sleep time was improved in the enzyme digested extracts as compared with the control group.
실시예Example 8: 8: 둥굴레Dingle 유기 용매 추출물의 제조 및 유기 용매 농도에 따른 수면 증진 기능 분석 Preparation of Organic Solvent Extract and Analysis of Sleep Enhancement Function by Organic Solvent Concentration
열수 추출을 대신하여 유기 용매의 추출 조건에 따른 수면 증진 효과를 확인하기 위하여 층층갈고리둥굴레 에탄올 추출물을 제조하였다. 먼저 추출 용매인 에탄올의 농도별 효과를 확인하기 위하여 층층갈고리둥굴레 뿌리 줄기 200g에 각각 10배의 40 ~ 70% 에탄올을 가한 후 60에서, 4시간 환류 추출하였다. 상기 환류 추출액을 원심분리(8,000 rpm, 20 min)한 후 농축기를 이용하여 용매를 모두 휘발시켰다. 이를 진공 가온 농축 후 동결건조하여 실시예 1의 방법에 따라 실시예 5의 절차에 따라 수면 증진 효과를 분석하였다. 본 실시예에 따른 분석 결과를 표 5에 나타낸다. 정상대조군 (CON)에 비해 에탄올의 함량이 증가할수록 수면 및 수면시간 중 Non-REM 시간이 유의적으로 증가하였다. Instead of hot water extraction, ethanol extracts of layered kangwon donggulle were prepared to investigate the effects of organic solvent extraction on the water. In order to investigate the effect of ethanol concentration on extracting solvent, ethanol extracts were prepared by adding 10 - fold of 40 ~ 70% ethanol to 200g of layered hooks, and then refluxed at 60 for 4 hours. The reflux extract was centrifuged (8,000 rpm, 20 min) and the solvent was evaporated using a concentrator. This was concentrated by vacuum heating and then lyophilized to analyze the effect of the water surface enhancement according to the procedure of Example 5 according to the method of Example 1. Table 5 shows the analysis results according to this embodiment. As the ethanol content increased, the non-REM time was significantly increased in the sleep and sleeping time compared to the normal control (CON).
농도(%)ethanol
density(%)
실시예Example 9: 9: 둥굴레Dingle 종별 및 추출 농도에 따른 수면 증진 기능 Sleep enhancement function by type and extraction concentration
둥굴레 속에 속하는 식물의 수면 증진 효능을 확인하기 위하여, 실시예 1(층층갈고리둥굴레), 실시예 2(대잎둥굴레), 실시예 3~4(기타 둥굴레) 에서 각각 제조된 둥굴레 추출물의 수면 증진 기능을 실시예 5의 절차에 따라 분석하였다. 본 실시예에 따른 분석 결과를 도 4에 나타낸다. 층층갈고리둥굴레 추출물은 대조군에 비하여 수면잠복기가 가장 크게 감소하고 총 수면시간이 가장 크게 증가하였다. 대잎둥굴레 추출물의 경우, 수면잠복기는 층층갈고리둥굴레 추출물과 동일 수준으로 감소하였으며, 총 수면 시간은 대조군에 비하여 약간 증가하였다. 잠정적으로 둥굴레 종으로 동정된 둥굴레 추출물은 수면 잠복기와 총 수면 시간이 대조군에 비하여 유의적인 차이를 보였으나 층층갈고리둥굴레 추출물에 비하면 낮은 수준으로 확인되었다. 따라서 여러 둥굴레 속 근경 추출물 중에 층층갈고리둥굴레 추출물의 효능이 가장 높은 것을 확인 할 수 있었다.In order to confirm the effect of improving the sleeping ability of the plants belonging to the genus Drosophilus, the water-improving function of the Drosophila extracts prepared in Example 1 (layered hooks), Example 2 (Example 1) and Examples 3 to 4 (other dongulla) And analyzed according to the procedure of Example 5. The results of the analysis according to this embodiment are shown in Fig. In the layered gill extracts, the latency of sleep was the greatest and the total sleep time was the greatest when compared with the control group. In the case of the extracts of Daburi extract, the latent period of sleep decreased to the same level as that of the layered hook, and total sleep time was slightly increased compared to the control group. The latent period and total sleep time of Dangguul extract, which was temporarily identified as Dangguul, were significantly lower than those of the control group, but lower than those of the layered duck extract. Therefore, it was confirmed that the extracts of Dahlgulrae extracts of layered roots were the most effective.
실시예Example 10: 방사선 동위원소 [ 10: Radiation isotope [ 33 H] 표지법을 통한 H] marking 둥굴레Dingle 추출물의 Extract GABAGABA AA 수용체 결합 능력 확인 Confirm receptor binding ability
실시예 1에서 제조된 층층갈고리둥굴레 추출물의 GABA A-benzodiazepine receptor의 결합능력을 확인하기 위하여 방사선 동위원소인 [3H] 표지법을 이용하였다. 머리를 절단한 Rat의 뇌의 대뇌피질 조직으로부터 GABA A-benzodiazepine receptor 표본을 만들고 둥굴레 추출물을 농도 별로 방사선동위원소가 포함된 [3H] flumazenil로 처리한 후 둥굴레 추출물을 농도 별로 처리하여 IC50([3H] flumazenil이 receptor에 절반이 결합한 상태)을 측정하여 benzodiazepine(BDZ)와 binding site에 대한 결합 경향성을 분석하였다. 구체적인 실험 방법은 다음과 같다.[ 3 H] labeling, a radioactive isotope, was used to confirm the binding ability of the GABA A-benzodiazepine receptor of the layered hookworm extract of Example 1. A process from the cortical tissue of the brain of the Rat a cutting head with GABA A-benzodiazepine receptor to create a sample containing the radioisotope at different concentrations of Polygonatum extract [3 H] flumazenil and then processes the Polygonatum extract by concentration IC 50 ( [ 3 H] flumazenil was bound to half of the receptor), and the binding tendency to benzodiazepine (BDZ) and binding site was analyzed. Specific experimental methods are as follows.
2 마리씩 SD rat (200-250 g)의 cerebral cortex를 10 mL Tris-HCl buffer(30 mM, pH 7.4)에 넣고 tissulyser를 이용하여 bead 넣어 10초간 homogenize 시켰다. 초원심분리(Ultra centrifuge; 48,000 rpm, 15분, 4)를 진행하고, 상층액을 버리고, pellet에 Tris-HCl buffer 10 mL을 넣어 세척(centrifuge 48,000 rpm, 15분)을 3회 반복 진행하였다. 37 water bath에서 30분간 incubation시켜 endogenous GABA 제거하였다. 새로운 centrifuge tube에 초원심분리(48,000 rpm, 10분, 4)를 진행하고, 상층액을 버리고, pellet에 Tris-HCl buffer 10 mL을 넣어 세척(centrifuge 48,000 rpm, 15분)을 진행하였다. 다시 상층액을 버리고 pellet에 Tris-HCl buffer(50 mM, pH 7.4) 15 mL을 넣고, binding시키기 전까지 -80에서 보관하였다(1-15일 보관 가능). 이를 녹인 후 tis-citrate buffer (50 mM, pH 7.1, 0-4) 1 mL씩 소분한 sample 5개(총 5 mL) 당 10 mL씩 분주하여 초원심분리(48,000 rpm, 10분, 4)를 2회 반복하였다. BCA(Bicinchoninic acid)를 측정하여 100 ㎍의 protein에 맞춰서 사용하였다. Two cerebellar cortices of SD rats (200-250 g) were placed in 10 mL Tris-HCl buffer (30 mM, pH 7.4) and homogenized for 10 seconds in a bead using a tissue syringe. The supernatant was discarded, and 10 mL of Tris-HCl buffer was added to the pellet, followed by washing (centrifugation at 48,000 rpm, 15 minutes) for 3 times. The supernatant was centrifuged at 48,000 rpm for 15 min. 37 water bath for 30 min to remove endogenous GABA. The supernatant was centrifuged (48,000 rpm, 10 min, 4) in a new centrifuge tube, and the supernatant was discarded. 10 mL of Tris-HCl buffer was added to the pellet and centrifuged at 48,000 rpm for 15 min. The supernatant was discarded and 15 mL of Tris-HCl buffer (50 mM, pH 7.4) was added to the pellet and stored at -80 ° C until binding. After dissolving it, 10 mL of 5 samples (5 mL total) was subdivided into 1 mL of tis-citrate buffer (50 mM, pH 7.1, 0-4) and centrifuged at 48,000 rpm for 10 min. And repeated twice. BCA (bicinchoninic acid) was measured and used in accordance with 100 μg of protein.
1) CON(total binding = receptor 329 ㎕ + tris-HCl buffer 150 ㎕ + radio-ligand 21 ㎕); 2) NSB (non-specific binding = receptor 329 ㎕ + BDZ(10 ㎎/㎖) 100 ㎕ + tris-HCl buffer 125 ㎕ + radio-ligand 21 ㎕); 3) H10 (receptor 329 ㎕ + H(10 ㎎/㎖) 25 ㎕ + tris-HCl buffer 125 ㎕ + radio-ligand 21 ㎕); 4) H50 (receptor 329 ㎕ + H(50 ㎎/㎖) 25 ㎕ + tris-HCl buffer 125 ㎕ + radio-ligand 21 ㎕); 총 500 ㎕, 3H의 final concentration 0.84 nM), 둥굴레 추출물 sample 총 5가지 농도 (0.01, 0.1, 1, 10, 100 ㎎/㎖)를 측정하였다. 0-4(냉장고) 에서 40분간 incubation 진행하고, GF/B glass fiber filter로 필터링을 진행하였다. 다시 0-4(냉장고)에서 overnight한 뒤에, LSC (Liquid Scintillation Counter) 이용하여 하기 식 (1)에 따라 결합 정도를 측정하였다. 1) CON (total binding = receptor 329 l + tris-HCl buffer 150 l + radio-ligand 21 l); 2) NSB (non-specific binding = receptor 329 μl + BDZ (10 mg / ml) 100 μl + tris-HCl buffer 125 μl + radio-ligand 21 μl); 3) H10 (25 μl of receptor 329 μl + H (10 mg / ml) + 125 μl of tris-HCl buffer + 21 μl of radio-ligand); 4) H50 (25 μl of receptor 329 μl + H (50 mg / ml) + 125 μl of tris-HCl buffer + 21 μl of radio-ligand); Total concentration of 500 ㎕, final concentration of 3H of 0.84 nM), and five concentrations (0.01, 0.1, 1, 10, 100 ㎎ / ㎖) Incubation was carried out in 0-4 (refrigerator) for 40 minutes, and filtration was performed with a GF / B glass fiber filter. After overnight at 0-4 (refrigerator), the degree of binding was measured according to the following equation (1) using LSC (liquid scintillation counter).
식 (1)에서, PSE는 둥굴레 추출물, DPM은 disintegration per minute을 의미한다. total binding DPM은 수용체에 동위원소만 넣어 결합 부위에 동위원소가 전부 결합할 때 측정되는 값을 의미한다. non specific binding DPM은 과량의 benzodiazepine과 함께 동위원소를 처리하였을 때 수용체의 결합 부위에 benzodiazepine이 붙고 다른 specific하지 않은 부분에 동위원소가 결합되는 값을 측정하여 total binding 에서 non specific binding 한 값을 빼서 계산한다. 식 (1)에 따라, 최종적으로 동위원소가 수용체가 결합하는 [3H] Binding (%)를 구할 수 있다. 이어서, 하기 식 (2)에 따라 동위원소를 대신하여 둥굴레 추출물이 어느 정도 수용체에 결합한 상대 값을 측정하고, 이를 그래프로 도시하였다. In Equation (1), PSE means dongguloe extract and DPM means disintegration per minute. Total binding DPM is the value measured when all isotopes are bound to the binding site with only the isotope at the receptor. Non-specific binding DPM is the result of binding of benzodiazepine to the binding site of the receptor when an isotope is treated with an excess of benzodiazepine and measuring the value of binding of the isotope to another unspecified part of the receptor, do. According to Eq. (1), finally [ 3 H] Binding (%) in which the isotope binds to the receptor can be obtained. Next, the relative value of the donguloe extract to the receptor to some extent in place of the isotope according to the following formula (2) was measured and shown in a graph.
전술한 방법에 따라, [3H]-flumazenil방법을 GABAA-benzodiazepine receptor binding assay를 진행하였으며, 측정 결과를 도 5에 나타낸다. 실험에 사용한 둥굴레 추출물의 농도는 100 ㎎/㎖에서 0.01 ㎎/㎖까지 5 구간으로 설정하였으며, 방사선 동위원소인 [3H]-flumazeni l(Ro15-1788) binding의 억제활성을 분석하였다. 둥굴레 추출물(PSE)은 중추성 benzodiazepine 수용체의 antagonist인 [3H]-flumazenil의 수용체 결합을 5구간의 농도에서 60.30 ~ 6.25%의 억제 활성도를 나타내어 농도 의존적으로 억제하였으며, IC50 값은 0.8152 ㎎/㎖로 확인되었다.According to the method described above, the [ 3 H] -flumazenil method was carried out on the GABAA-benzodiazepine receptor binding assay, and the measurement results are shown in FIG. The concentrations of the extracts of Dangguul extracts were set to 5 sections from 100 ㎎ / ㎖ to 0.01 ㎎ / ㎖ and the inhibitory activity of the radioactive isotope [3H] - flumazeni l (Ro15-1788) binding was analyzed. The inhibition of receptor binding of [3H] -flumazenil, an antagonist of the central benzodiazepine receptor, was inhibited by 60.30 ~ 6.25% concentration at 5 concentration, and the IC 50 value was 0.8152 ㎎ / ㎖ Respectively.
제조예Manufacturing example : : 둥굴레Dingle 추출물 함유 조성물 제조 Preparation of extract-containing composition
다음과 같은 조성을 가지는 타정 제형을 가지는 조성물을 제조하였다. 실시예 1에서 얻은 둥굴레 물 추출 분말 500 ㎎, 결정 셀룰로오스 88 ㎎, 말토덱스트린 40 ㎎, 유당 혼합 분말 129 ㎎, 스테아린산 마그네슘 10 ㎎, 히드록시프로필메틸셀룰로오스(HPMC) 21 ㎎. A composition having a tablet formulation having the following composition was prepared. 500 mg of water extract of Sambrooki obtained in Example 1, 88 mg of crystalline cellulose, 40 mg of maltodextrin, 129 mg of lactose mixed powder, 10 mg of magnesium stearate, and 21 mg of hydroxypropyl methylcellulose (HPMC).
상기에서는 본 발명의 예시적인 실시형태 및 실시예에 기초하여 본 발명을 설명하였으나, 본 발명이 상기 실시형태 및 실시예에 기재된 기술사상으로 한정되지 않는다. 오히려 본 발명이 속하는 기술분야에서 통상의 지식을 가진 자라면 전술한 실시형태 및 실시예를 토대로 다양한 변형과 변경을 용이하게 추고할 수 있다. 하지만, 이러한 변형과 변경은 모두 본 발명의 권리범위에 속한다는 점은, 첨부하는 청구범위에서 분명하다.Although the present invention has been described based on the exemplary embodiments and examples of the present invention, the present invention is not limited to the technical ideas described in the above embodiments and examples. Those skilled in the art will appreciate that various modifications and changes may be made without departing from the spirit and scope of the present invention as defined by the appended claims. It is apparent, however, that the appended claims are intended to cover all such modifications and changes as fall within the true scope of the invention.
Claims (10)
A pharmaceutical composition for treating sleep disorders containing Polygonatum falcatum hot water extract as an active ingredient.
상기 대잎둥굴레 열수 추출물은 상기 약학 조성물 중에 0.01 ~ 1000 ㎎/㎖의 농도로 함유되는 수면 장애를 치료하기 위한 약학 조성물.
The method according to claim 1,
The pharmaceutical composition for treating sleep disorders, wherein the water extract of Daegu Dangguljang is contained in the pharmaceutical composition at a concentration of 0.01 to 1000 mg / ml.
A functional food composition for enhancing sleeping time and improving sleep quality containing hot water extract of Polygonatum falcatum as an active ingredient.
상기 대잎둥굴레 열수 추출물은 상기 기능성 식품 조성물 중에 0.01 ~ 1000 ㎎/㎖의 농도로 함유되는 수면 시간 증진 및 수면 질 개선용 기능성 식품 조성물.
The method of claim 3,
The functional food composition for enhancing sleeping time and improving the quality of sleeping water comprises the above-mentioned water extract of Daegu Dangguljoo at a concentration of 0.01 to 1000 mg / ml in the functional food composition.
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