KR20190040170A - A Pharmaceutical Composition comprising Arctium lappa root extract for depression or stress relief, and a composition for improving cognitive function - Google Patents
A Pharmaceutical Composition comprising Arctium lappa root extract for depression or stress relief, and a composition for improving cognitive function Download PDFInfo
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- KR20190040170A KR20190040170A KR1020190041994A KR20190041994A KR20190040170A KR 20190040170 A KR20190040170 A KR 20190040170A KR 1020190041994 A KR1020190041994 A KR 1020190041994A KR 20190041994 A KR20190041994 A KR 20190041994A KR 20190040170 A KR20190040170 A KR 20190040170A
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- depression
- extract
- present
- burdock
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Abstract
Description
본 발명은 우엉 추출물을 포함하는 우울증 또는 스트레스 완화, 및 인지기능 개선용 조성물에 관한 것이다. 보다 상세하게는, 본 발명은 우엉 추출물을 포함하는 우울증 예방 및 치료용 조성물, 우엉 추출물을 포함하는 스트레스 완화용 조성물, 및 우엉 추출물을 포함하는 인지기능 개선용 조성물에 관한 것이다.The present invention relates to a composition for improving depression or stress alleviation, and cognitive function comprising burdock extract. More particularly, the present invention relates to a composition for preventing and treating depression comprising burdock extract, a composition for relieving stress including burdock extract, and a composition for improving cognitive function including burdock extract.
스트레스는 외부 자극에 의해 발생되는 유기체에 대한 해로운 반응 또는 변화를 의미한다. 일반적으로, 생체에 미치는 자극이 일정한 정도 이상이 될 경우 상해적으로 작용하는데, 이때 생체는 그 자극의 종류에 관계없이 일정한 위협에 대한 반응으로 긴장성 두통, 편두통, 고혈압, 소화불량, 피로, 통증, 발모 및 피부의 거칠어짐 등을 일으키고, 만성적으로 지속되면 각종 신경증, 위궤양 등을 유발시키는 등의 비특이적인 전신적증후군(general adaptive syndrome)을 나타내게 되는데, 이를 스트레스라 정의하고 있다(Selye, 1958, Longmans Green and Co.). 최근 여러 보고에 의하면, 질병의 90% 이상이 스트레스에 기안한다고 알려져 있다. 최근 우리나라 보건복지부에서도 스트레스를 새로운 질병코드로 분류하고 있다. 특히, 우울증과 같은 정신장애, 신경성위, 십이지장궤양 등과 같은 소화기장애, 스트레스성 갑상선기능항진증과 같은 내분비장애, 스트레스성 기미나 여드름 등과 같은 피부장애, 및 부정맥이나 협심증과 같은 심혈관계 장애 등의 다양한 질환이 스트레스로 인해 유발되는 것으로 보고된바 있다.Stress means a harmful reaction or change to an organism caused by external stimuli. In general, when a stimulus on a living body becomes more than a predetermined level, the living body works harmlessly. In this case, a living body responds to a certain threat irrespective of the type of the stimulus, (Selye, 1958, Longmans Green, 1986). In the present study, we used the term "stress" to refer to stress syndrome, which is characterized by a generalized adaptive syndrome that causes hair growth and skin roughness, and chronic persistence of various neuroses and gastric ulcers and Co.). According to recent reports, it is known that more than 90% of diseases are caused by stress. Recently, the Ministry of Health and Welfare has classified stress into a new disease code. Particularly, there are various kinds of psychological disorders such as mental disorders such as depression, digestive disorders such as nerve stomach and duodenal ulcer, endocrine disorders such as stressful hyperthyroidism, skin disorders such as stressed spots and acne, and cardiovascular disorders such as arrhythmia and angina The disease has been reported to be caused by stress.
우울증은 객관적 상황과는 관계없이 일어나는 정서적 병리현상으로 환자의 모든 생활이 우울한 기분으로 덮여있고, 흥미가 감소하고 무쾌감증(anhedonia)이 되며 정신운동의 저하, 염세감, 절망에 사로잡히게 되고 자살의욕을 느껴 자살기도에까지 이르는 질병인데, 식욕저하, 불면, 변비, 성욕감퇴 등 다양한 신체적 증상을 보인다. 이러한 우울증은 최근 심각한 사회적 문제로 대두되고 있다.Depression is an emotional pathology that occurs regardless of the objective situation. All lives of the patient are covered with depressed moods, decreased in interest, become anhedonia, become depressed, depressed, and desperate. It is a disease that reaches to the suicide prayer because it is motivated and it shows various physical symptoms such as loss of appetite, insomnia, constipation, loss of libido. Such depression has become a serious social problem in recent years.
한편, 전체 여성의 20% 정도가 그 증상을 겪는 것으로 알려진 갱년기는, 노화 현상의 하나로, 난소의 기능이 점차 감소하는 40세 이후부터 서서히 시작되는데, 여성의 평균 수명이 점차 연장됨에 따라 호르몬 결핍기간이 점차 길어지고 있는 점이 문제점으로 대두되고 있다. 난소 기능의 감소, 즉 에스트로겐 감소로, 안면홍조, 발한, 불면증, 신경과민, 우울증, 현기증, 집중력 결핍, 단기 기억장애, 불안, 기억력 감퇴, 심계항진, 근육통, 관절통, 피부건조, 질건조증, 질위축, 하부 요도 위축, 질염, 자궁위축, 방광염, 배뇨통, 급뇨, 복부비만, 고지혈증, 동맥경화 및 지방간 등의 신체 및 정신적인 변화가 나타난다. 이러한 증상 중에서도 여성 갱년기 우울증은 삶의 질과 직접적으로 연관되므로 보다 심각한 문제로 대두되고 있다.On the other hand, menopause, which is known to cause 20% of all women to experience the symptoms, starts gradually after the age of 40, when the function of the ovaries declines gradually. As the life expectancy of women gradually increases, Which is becoming more and more problematic. It has been shown that ovarian function is reduced, that is, oestrogen is reduced, resulting in facial flushing, sweating, insomnia, nervousness, depression, dizziness, concentration deficiency, short term memory impairment, anxiety, memory loss, palpitations, muscle pain, arthralgia, , Lower urinary tract atrophy, vaginitis, uterine atrophy, cystitis, dysuria, diaphoresis, abdominal obesity, hyperlipidemia, arteriosclerosis and fatty liver. Among these symptoms, women's menopausal depression is directly related to the quality of life, so it is becoming more serious problem.
또한, 우울증은 동물에서도 나타날 수 있는데, 최근 반려동물에 대한 관심이 높아지면서 이들의 삶의 질을 함께 개선하고자 하는 인식이 확산되고 있다. 따라서, 동물의 우울증을 치료하기 위한 방법 또한 높은 관심을 받고 있다. 동물의 경우 우울증을 치료, 개선하기 위한 약제학적 조성물 외에 기능성 사료 첨가제 조성물로도 적용될 수 있다.Depression can also occur in animals. As interest in companion animals has increased in recent years, there is a growing awareness of improving their quality of life. Thus, methods for treating depression in animals have also received high interest. In the case of animals, a pharmaceutical composition for treating or ameliorating depression may be applied as a functional feed additive composition.
현재의 항우울제는 대부분 중추 세로토닌 또는 노르아드레날린 시냅스에서 신경전달물질의 농도를 높이는 약리작용을 가지고 있다. 항우울제는 신경전달물질의 농도를 높여주는 메커니즘에 따라 크게 삼환계 항우울제(TCA; tricyclic antidepressants), 모노아민 옥시다제 억제제(MAOI; monoamine oxidase inhibitors), 또는 선택적 세로토닌 재흡수 억제제(SSRI; selective serotonin reuptake inhibitors) 등이 많이 사용되고 있다.Current antidepressants have a pharmacological action that increases the concentration of neurotransmitters in central serotonin or noradrenaline synapses. Antidepressants can be classified as tricyclic antidepressants (TCA), monoamine oxidase inhibitors (MAOI), or selective serotonin reuptake inhibitors (SSRIs) depending on the mechanisms that increase the concentration of neurotransmitters. And so on.
이러한 기존의 항우울제가 비교적 효과적이기는 하나, 일부에서 심한 부작용 및 기대에 미치지 못하는 효과가 나타나는 경우도 있다. 항우울증 약물에 잘 반응하지 않거나(Thase et al. 2001), 재발률이 높으며(Frank et al. 1991), 심한 부작용에 의해 약물복용이 불가능한 환자들도 많다(Gumnick and Nemeroff 2000). 삼환계 항우울제(tricyclic antidepressants: TCA)인 이미프라민(imipramine), 데시프라민(desipramine)등은 저혈압, 심장기능장애 등의 부작용이 심하게 나타난다. 가장 널리 쓰이는 플루아크서틴 (fluoxetine)이나 서트랄린(sertraline) 등의 선택적 세로토닌 재흡수 억제제는 구역, 위장관 출혈 또는 성기능 장애 등을 유발한다.Although these existing antidepressants are relatively effective, some of them may have serious side effects and effects that do not meet expectations. (Geminick and Nemeroff, 2000). Inadequate response to antidepressant drugs (Thase et al., 2001), high recurrence rates (Frank et al. Tricyclic antidepressants (TCA) imipramine, desipramine, etc., have severe side effects such as hypotension and cardiac dysfunction. The most commonly used selective serotonin reuptake inhibitors, such as fluoxetine and sertraline, cause zone, gastrointestinal bleeding or sexual dysfunction.
한편, 노인성 치매의 일종인 알츠하이머병으로부터 유발되는 기억력 및 학습능력의 상실 등 인지 기능 장애는 대뇌 기저부의 아세틸콜린성 신경세포의 손상에서 비롯된다는 연구결과에 의해 무스카린성 아세틸콜린 수용체(muscarine acetylcholine receptor)에 대한 효능제(agonist), 아세틸콜린 생성촉진제, 아세틸콜린에스터라제 저해제 등 여러 가지 작용기전에 따라 아세틸콜린성 신경세포의 기능을 강화시켜줄 수 있는 약물들이 개발되어 왔다. 현재 아세틸콜린에스터라제 저해제로 타크린(tacrine)이 개발되어 있으나 위장장애와 같은 일부 부작용이 나타나고 있어 천연 식물소재로부터 저해활성소재를 탐색하여 활용할 필요가 있다.On the other hand, research has shown that cognitive dysfunction, such as loss of memory and learning ability caused by Alzheimer's disease, a kind of senile dementia, results from damage to the acetylcholinergic neurons of the cerebral basal ganglia, resulting in muscarine acetylcholine receptors Drugs capable of enhancing the function of acetylcholinergic neurons in response to various functional groups such as agonists, acetylcholine production promoters, and acetylcholinesterase inhibitors have been developed. Although tacrine has been developed as an acetylcholinesterase inhibitor, some side effects such as gastrointestinal disorders are present and it is necessary to search for inhibitory active materials from natural plant materials.
우엉은 국화과 두해살이풀로, 유럽 원산의 귀화식물이다. 높이는 1.5m정도이며, 뿌리는 30-60 ㎝ 이다. 우엉은 알칼리성 식품으로 특유한 향기가 있으며, 뿌리에는 이눌린과 약간의 팔미트산이 포함되어 있고, 이뇨제 또는 발한제로 사용된다. 특히 종자는 우방자라 하여 부기가 있을 때 이뇨제로 사용하며, 인후통과 독충(毒蟲)의 해독제로 쓰인다. 한편, 우엉에는 항산화 작용을 하는 폴리페놀과 레스베라트롤이 풍부하게 함유되어 있고, 혈액순환을 돕고 상처를 치료하는 사포닌이 들어있어 뇌질환, 심장병, 염증에 효과가 있으며, 콜레스테롤 수치를 낮추어 변비를 예방하기 때문에 다이어트에 도움이 되는 것으로 알려져 있다.Burdock is a national flower and biennial plant, native to Europe. The height is about 1.5m and the root is 30-60cm. Burdock is an alkaline food with a characteristic scent, the root contains inulin and some palmitic acid, and is used as a diuretic or sweating agent. Especially, seeds are used as diuretic when there is a bookkeeping due to the alien, and they are used as an antidote to sore throat and poisonous insect. On the other hand, burdock is rich in antioxidant polyphenols and resveratrol. It contains saponin that helps blood circulation and treats wounds, and is effective in brain diseases, heart disease, inflammation, and lowering of cholesterol levels to prevent constipation Because it is known to help with diet.
대한민국 등록특허 제10-1557716호는 우엉의 항비만 효과를 개시하고 있고, 대한민국 공개특허 제 10-2015-0051597호는 우엉의 간기능 개선 효과를 개시하고 있지만, 우울증 또는 인지기능 개선과 관련된 효과는 보고된 바 없다. 이에 본 발명자들은 상술한 부작용없이 우울증 치료 또는 인지기능 개선에 효과적인 우엉 추출물을 포함하는 조성물을 제공하고자 한다.Korean Patent No. 10-1557716 discloses an anti-obesity effect of burdock, Korean Patent Publication No. 10-2015-0051597 discloses an effect of improving burdock liver function, but the effect associated with depression or cognitive function improvement Not reported. Accordingly, the present inventors intend to provide a composition containing a burdock extract effective for treating depression or improving cognitive function without the above-mentioned side effects.
본 발명자들은 기존의 항우울제에서 나타나는 부작용을 개선하기 위해 천연추출물을 이용한 우울증 치료제를 개발하고자 예의 연구 노력하였다. 그 결과, 본 발명자들은 우엉 추출물이 우울증의 치료 또는 개선에 효과가 있음을 확인하였으며, 특히 우엉 추출물이 스트레스 또는 우울증 증상으로 인해 감소하는 BDNF (brain derivated neurotrophic factor) 단백질 발현량을 정상수준으로 회복시킴을 확인함으로써 본 발명을 완성하게 되었다.The present inventors have tried to develop a therapeutic agent for depression using natural extracts in order to improve the side effects of conventional antidepressants. As a result, the present inventors confirmed that burdock extract was effective for the treatment or improvement of depression, and particularly recovered the amount of brain derived neurotrophic factor (BDNF) protein expressed by stress or depression symptoms to normal levels Thereby completing the present invention.
따라서, 본 발명의 목적은 우울증 예방 또는 치료용 약제학적 조성물을 제공하는 데 있다.Accordingly, it is an object of the present invention to provide a pharmaceutical composition for preventing or treating depression.
본 발명의 다른 목적은 우울증 개선 또는 예방용 식품 조성물을 제공하는 데 있다.It is another object of the present invention to provide a food composition for improving or preventing depression.
본 발명의 또 다른 목적은 인지기능 장애 예방 또는 치료용 약제학적 조성물을 제공하는 데 있다.It is another object of the present invention to provide a pharmaceutical composition for preventing or treating cognitive dysfunction.
본 발명의 또 다른 목적은 인지능력 또는 기억력 개선 식품 조성물을 제공하는 데 있다.It is another object of the present invention to provide a cognitive or memory-improving food composition.
본 발명의 또 다른 목적은 여성 갱년기 증후군 예방 또는 치료용 약제학적 조성물 및 여성 갱년기 증후군 개선 또는 예방 식품 조성물을 제공하는 데 있다.It is another object of the present invention to provide a pharmaceutical composition for preventing or treating female menopausal syndrome and a composition for improving or preventing female menopausal syndrome.
본 발명의 또 다른 목적은 우울증 또는 스트레스 개선 또는 예방 기능성 사료 첨가제 조성물, 및 동물 인지능력 향상 또는 개선 기능성 사료 첨가제 조성물을 제공하는 데 있다.It is another object of the present invention to provide a depression or stress-improving or preventive functional feed additive composition, and an animal cognition enhancing or improving functional feed additive composition.
본 발명의 또 다른 목적은 항스트레스 조성물을 제공하는 데 있다.It is another object of the present invention to provide an anti-stress composition.
본 발명의 일 양태에 따르면, 본 발명은 (a) 우엉(Arctium lappa root) 추출물의 치료학적 유효량 및 (b) 약제학적으로 허용되는 담체를 포함하는 우울증 예방 또는 치료용 약제학적 조성물을 제공한다.According to one aspect of the present invention, there is provided a pharmaceutical composition for the prevention or treatment of depression comprising (a) a therapeutically effective amount of an extract of Arctium lappa root and (b) a pharmaceutically acceptable carrier.
본 발명자들은 기존의 항우울제에서 나타나는 부작용을 개선하기 위해 천연추출물을 이용한 우울증 치료제를 개발하고자 예의 연구 노력한 결과, 우엉 추출물이 우울증의 치료 또는 개선에 효과가 있음을 확인하였으며, 특히 우엉 추출물이 스트레스 또는 우울증 증상으로 인해 감소하는 BDNF (brain derivated neurotrophic factor) 단백질 발현량을 정상수준으로 회복시킴을 확인하였다.The inventors of the present invention have made efforts to develop a therapeutic agent for depression using natural extracts in order to improve adverse effects in existing antidepressants. As a result, it was confirmed that burdock extract was effective in the treatment or improvement of depression, And the amount of BDNF (brain derived neurotrophic factor) protein expression, which is decreased due to symptoms, is restored to a normal level.
본 발명에서 이용되는 우엉 추출물은 구입하거나 또는 직접 추출하여 얻을 수 있다. 본 발명의 조성물에서 이용되는 우엉 추출물을 우엉에 추출용매를 처리하여 얻는 경우에는, 다양한 추출용매가 이용될 수 있으며, 예컨대, 극성 용매 또는 비극성 용매를 이용할 수 있다. 극성 용매로서 적합한 것은, (i) 물, (ii) 알코올(바람직하게는, 메탄올, 에탄올, 프로판올, 부탄올, 노말-프로판올, 이소-프로판올, 노말-부탄올, 1-펜탄올, 2-부톡시에탄올 또는 에틸렌글리콜), (iii) 아세트산, (iv) DMFO(dimethyl-formamide) 및 (v) DMSO(dimethyl sulfoxide)를 포함한다. 비극성 용매로서 적합한 것은, 아세톤, 아세토나이트릴, 에틸 아세테이트, 메틸 아세테이트, 플루오로알칸, 펜탄, 헥산, 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, 어닐린, 디에틸아민, 에테르, 사염화탄소 및 THF(Tetrahydrofuran)를 포함한다.The burdock extract used in the present invention can be obtained by purchase or by direct extraction. When the extract of burdock root used in the composition of the present invention is obtained by treating an extraction solvent in burdock, various extraction solvents may be used. For example, a polar solvent or a non-polar solvent may be used. Suitable polar solvents are (i) water, (ii) alcohols (preferably methanol, ethanol, propanol, butanol, n-propanol, iso-propanol, n-butanol, 1-pentanol, Or ethylene glycol), (iii) acetic acid, (iv) dimethyl-formamide (DMFO) and (v) dimethyl sulfoxide (DMSO). Suitable nonpolar solvents are acetone, acetonitrile, ethyl acetate, methyl acetate, fluoroalkane, pentane, hexane, 2,2,4-trimethylpentane, decane, cyclohexane, cyclopentane, diisobutylene, 1- But are not limited to, pentane, 1-chlorobutane, 1-chloropentane, o-xylene, diisopropyl ether, 2- chloropropane, toluene, 1- chloropropane, chlorobenzene, benzene, diethyl ether, diethylsulfide, Methane, 1,2-dichloroethane, aniline, diethylamine, ether, carbon tetrachloride and THF (Tetrahydrofuran).
본 발명의 일 구현예에 따르면, 본 발명에서 이용되는 우엉 추출물은 물, C1 내지 C4의 저급 알코올 또는 이들의 혼합용매에 의해 추출가능하다. 본 발명에 따르면, 본 발명의 조성물은 에틸알코올을 용매로 한 우엉 주정 추출물을 포함할 수 있다. 우엉 추출물은 우엉을 우엉 무게(g) 10 배수 부피(ml)의 추출용매에 투입한 후 진공저온 추출장치를 사용하여 70-90℃에서 5-7시간 동안 추출하여 수득할 수 있다. 본 발명의 일 예에 따르면, 우엉 추출물은 우엉을 추출용매에 투입한 후 75-85℃에서 5.5-6.5시간 동안 추출하여 수득할 수 있다. 본 발명의 특정 구현예에 따르면, 우엉 추출물을 직접 추출하는 방법의 예는 다음과 같다. 우엉 600 g을 10 배수의 주정 6 L에 투입한 후 진공저온 추출장치(COSMOS-660)를 사용하여 80℃에서 6시간 동안 추출하였다. 추출된 농축액을 동결 건조하여 우엉 추출 분말을 얻었다. 본 발명에서 우엉 추출물은 용매에 의해 추출된 crude 형태를 이용할 수 있으며, 고순도로 정제하여 사용할 수도 있다.According to one embodiment of the present invention, the burdock extract used in the present invention can be extracted with water, C1 to C4 lower alcohols or a mixed solvent thereof. According to the present invention, the composition of the present invention may comprise a burdock alcohol extract in which ethyl alcohol is used as a solvent. The burdock extract can be obtained by charging the burdock to burdock weight (g) 10 times the volume (ml) of the extraction solvent and then extracting it at 70-90 ° C for 5-7 hours using a vacuum low temperature extraction apparatus. According to one example of the present invention, burdock root extract can be obtained by adding burdock to an extraction solvent and then extracting it at 75-85 ° C for 5.5-6.5 hours. According to a specific embodiment of the present invention, an example of a method for directly extracting burdock root extract is as follows. 600 g of burdock was added to 6 L of a 10-fold aliquot, which was then extracted with a vacuum low-temperature extraction apparatus (COSMOS-660) at 80 ° C for 6 hours. The extracted concentrate was lyophilized to obtain burdock extract powder. In the present invention, the burdock extract may be a crude form extracted with a solvent, and may be purified and used in high purity.
본 명세서에서 사용되는 용어 '추출물'은 상술한 바와 같이 당업계에서 조추출물(crude extract)로 통용되는 의미를 갖지만, 광의적으로는 추출물을 추가적으로 분획(fractionation)한 분획물도 포함한다. 즉, 우엉 추출물은 상술한 추출용매를 이용하여 얻은 것뿐만 아니라, 여기에 정제과정을 추가적으로 적용하여 얻은 것도 포함한다. 예컨대, 상기 추출물을 일정한 분자량 컷-오프 값을 갖는 한외 여과막을 통과시켜 얻은 분획, 다양한 크로마토그래피 (크기, 전하, 소수성 또는 친화성에 따른 분리를 위해 제작된 것)에 의한 분리 등, 추가적으로 실시된 다양한 정제 방법을 통해 얻어진 분획도 본 발명의 우엉 추출물에 포함되는 것이다.As used herein, the term " extract " means that it is used in the art as a crude extract as described above, but broadly includes fractions obtained by further fractionating the extract. That is, the burdock extract includes not only those obtained by using the aforementioned extraction solvent but also those obtained by further applying a purification process thereto. For example, a fraction obtained by passing the above extract through an ultrafiltration membrane having a constant molecular weight cut-off value, and a separation by various chromatography (manufactured for separation according to size, charge, hydrophobicity or affinity) The fraction obtained by the purification method is also included in the burdock extract of the present invention.
본 발명에서 이용되는 우엉 추출물은 감압 증류 및 동결 건조 또는 분무 건조 등과 같은 추가적인 과정에 의해 분말 상태로 제조될 수 있다.The burdock extract used in the present invention may be prepared in powder form by an additional process such as vacuum distillation and freeze drying or spray drying.
본 명세서에서 용어 '치료학적 유효량'이란 하기의 우엉의 효능 또는 활성을 달성하는 데 충분한 양을 포함하는 것을 의미한다. 본 발명은 천연 식물재료인 우엉으로부터 추출한 조성물로서 과량 투여하여도 인체에 부작용이 없으므로 우엉 추출물이 본 발명의 조성물에 포함된 양적 상한은 당업자가 적절한 범위 내에서 선택하여 실시할 수 있다.The term "therapeutically effective amount" as used herein is meant to include an amount sufficient to achieve the following efficacy or activity of the burdock. Since the present invention is a composition extracted from burdock which is a natural plant material, there is no adverse effect on the human body even when it is administered in an excessive amount, so that the quantitative upper limit of the burdock extract contained in the composition of the present invention can be selected by a person skilled in the art within a suitable range.
하기의 실시예에서 입증된 바와 같이, 우엉 추출물의 투여는 신경세포 보호효과를 나타내고, 강제 수영 실험에서 움직이는 시간(swimming time)이 증가하여 항우울 효과를 나타내었다. 이와 같은 실험 데이터는 본 발명의 조성물이 우울증의 예방 또는 치료에 이용될 수 있으며, 종래 항우울제의 대체제로 이용될 수 있음을 보여주는 것이다.As demonstrated in the following examples, the administration of burdock extracts exhibited neuroprotective effects and increased anti-depressive effects by increased swimming time in forced swimming experiments. Such experimental data show that the composition of the present invention can be used for the prevention or treatment of depression and can be used as a substitute for conventional antidepressants.
본 발명의 약제학적 조성물은 약제학적으로 허용되는 담체를 포함한다. 상기 약제학적으로 허용되는 담체는 제제시에 통상적으로 이용되는 것으로서, 락토스, 덱스트로스, 수크로스, 솔비톨, 만니톨, 전분, 아카시아 고무, 인산 칼슘, 알기네이트, 젤라틴, 규산 칼슘, 미세결정성 셀룰로스, 폴리비닐피롤리돈, 셀룰로스, 물, 시럽, 메틸 셀룰로스, 메틸히드록시벤조에이트, 프로필히드록시벤조에이트, 활석, 스테아르산 마그네슘 및 미네랄 오일 등을 포함하나, 이에 한정되는 것은 아니다. 본 발명의 약제학적 조성물은 상기 성분들 이외에 윤활제, 습윤제, 감미제, 향미제, 유화제, 현탁제, 보존제 등을 추가로 포함할 수 있다. 적합한 약제학적으로 허용되는 담체 및 제제는 Remington's Pharmaceutical Sciences(19th ed., 1995)에 상세히 기재되어 있다.The pharmaceutical composition of the present invention includes a pharmaceutically acceptable carrier. Such pharmaceutically acceptable carriers are those conventionally used in the field of manufacture and include lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia rubber, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, But are not limited to, polyvinylpyrrolidone, cellulose, water, syrup, methylcellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. The pharmaceutical composition of the present invention may further contain a lubricant, a wetting agent, a sweetening agent, a flavoring agent, an emulsifying agent, a suspending agent, a preservative, etc. in addition to the above components. Suitable pharmaceutically acceptable carriers and formulations are described in detail in Remington ' s Pharmaceutical Sciences (19th ed., 1995).
본 발명의 약제학적 조성물은 경구 또는 비경구로 투여할 수 있고, 비경구 투여인 경우에는 정맥내 주입, 피하 주입, 근육 주입, 복강 주입, 경피 투여 등으로 투여할 수 있으며, 바람직하게는 경구 투여이다.The pharmaceutical composition of the present invention can be administered orally or parenterally. In the case of parenteral administration, the composition can be administered by intravenous injection, subcutaneous injection, muscle injection, intraperitoneal injection, transdermal administration, etc., .
본 발명의 약제학적 조성물의 적합한 투여량은 제제화 방법, 투여방식, 환자의 연령, 체중, 성, 병적 상태, 음식, 투여 시간, 투여 경로, 배설 속도 및 반응 감응성과 같은 요인들에 의해 다양하며, 보통으로 숙련된 의사는 소망하는 치료 또는 예방에 효과적인 투여량을 용이하게 결정 및 처방할 수 있다. 본 발명의 바람직한 구현예에 따르면, 본 발명의 약제학적 조성물의 1일 투여량은 0.001-10000 ㎎/㎏이다.The appropriate dosage of the pharmaceutical composition of the present invention varies depending on factors such as the formulation method, administration method, age, body weight, sex, pathological condition, food, administration time, administration route, excretion rate and responsiveness of the patient, Usually, a skilled physician can readily determine and prescribe dosages effective for the desired treatment or prophylaxis. According to a preferred embodiment of the present invention, the daily dosage of the pharmaceutical composition of the present invention is 0.001-10000 mg / kg.
본 발명의 약제학적 조성물은 당해 발명이 속하는 기술분야에서 통상의 지식을 가진 자가 용이하게 실시할 수 있는 방법에 따라, 약제학적으로 허용되는 담체 및/또는 부형제를 이용하여 제제화 함으로써 단위 용량 형태로 제조되거나 또는 다용량 용기 내에 내입시켜 제조될 수 있다. 이때 제형은 오일 또는 수성 매질중의 용액, 현탁액 또는 유화액 형태이거나 엑스제, 분말제, 과립제, 정제 또는 캅셀제 형태일 수도 있으며, 분산제 또는 안정화제를 추가적으로 포함할 수 있다.The pharmaceutical composition of the present invention may be formulated into a unit dose form by formulating it using a pharmaceutically acceptable carrier and / or excipient according to a method which can be easily carried out by a person having ordinary skill in the art to which the present invention belongs. Or by intrusion into a multi-dose container. The formulations may be in the form of solutions, suspensions or emulsions in oils or aqueous media, or in the form of excipients, powders, granules, tablets or capsules, and may additionally contain dispersing or stabilizing agents.
본 발명의 다른 양태에 따르면, 본 발명은 (a) 우엉(Arctium lappa root) 추출물의 치료학적 유효량 및 (b) 약제학적으로 허용되는 담체를 포함하는 여성 갱년기 증후군 예방 또는 치료용 약제학적 조성물을 제공한다.According to another aspect of the present invention there is provided a pharmaceutical composition for the prevention or treatment of female menopausal syndrome comprising (a) a therapeutically effective amount of an extract of Arctium lappa root and (b) a pharmaceutically acceptable carrier do.
본 발명의 여성 갱년기 증후군 개선 또는 예방용 식품 조성물은 상술한 "우울증 예방 또는 치료용 약제학적 조성물"과 동일하게 유효성분으로서 우엉 추출물을 이용하기 때문에, 공통된 사항은 본 명세서의 과도한 중복성을 피하기 위해 그 기재를 생략한다.Since the food composition for improving or preventing female menopausal symptoms of the present invention uses burdock extract as an active ingredient in the same manner as the above-mentioned " pharmaceutical composition for preventing or treating depression ", a common point is that, in order to avoid excessive redundancy of the present specification The description is omitted.
여성의 갱년기 증상의 종류와 증세의 정도에는 개인차가 심하여, 일상생활에 지장이 없을 정도의 가벼운 경우에서 아주 거동이 불편하게 될 정도의 중증까지 다양하게 나타난다. 갱년기 여성의 약 30%가 심한 증상으로 치료를 받고 있는 것으로 알려져 있고, 치료제로는 여성호르몬 투여가 있으나 부작용에 대한 보고가 있으며, 지속적인 골밀도 감소로 인한 골다공증에 대한 치료가 필요한 경우도 있다.The type of menopausal symptom and severity of symptoms in women are severe, ranging from mild cases that do not interfere with daily life to severe conditions that are very uncomfortable. It is known that about 30% of menopausal women are treated with severe symptoms, and there are reports of adverse effects, although treatment with female hormones has been reported, and treatment for osteoporosis due to continuous reduction in bone density is sometimes necessary.
여성 갱년기 증후군에는 안면홍조, 발한, 불면증, 신경과민, 우울증, 현기증, 집중력 결핍, 단기 기억장애, 불안, 기억력 감퇴, 심계항진, 근육통, 관절통, 피부건조, 질건조증, 질위축, 하부 요도 위축, 질염, 자궁위축, 방광염, 배뇨통, 급뇨, 복부비만, 고지혈증, 동맥경화 또는 지방간 등이 있다. 본 발명의 일 구현예에 따르면, 상기 여성 갱년기 증후군은 불면증, 신경과민, 우울증, 현기증, 집중력 결핍, 단기 기억장애, 불안 및 기억력 감퇴로 구성된 군에서 선택되는 1 이상의 증상을 의미한다. 본 발명의 특정 구현예에 따르면, 상기 여성 갱년기 증후군은 우울증 또는 기억력 감퇴이다.Women's menopausal symptoms include facial flushing, sweating, insomnia, nervousness, depression, dizziness, impaired concentration, short-term memory impairment, anxiety, memory loss, palpitations, myalgia, arthralgia, vaginal dryness, vaginal atrophy, , Uterine atrophy, cystitis, dysuria, urination, abdominal obesity, hyperlipidemia, arteriosclerosis or fatty liver. According to one embodiment of the invention, the female menopausal syndrome means one or more symptoms selected from the group consisting of insomnia, nervousness, depression, dizziness, concentration deficiency, short-term memory impairment, anxiety and memory impairment. According to certain embodiments of the invention, the female menopausal syndrome is depression or memory loss.
갱년기 증상은 삶의 질과 직접적으로 연관되고 장기간 나타나기 때문에, 이의 지속적인 치료를 위한 부작용이 없는 천연물질 유래의 약학적 제제 또는 건강기능식품의 개발이 필요하다.Since menopausal symptoms are directly related to the quality of life and appear for long periods of time, it is necessary to develop pharmaceutical products or health functional foods derived from natural substances that have no adverse effects for their continued treatment.
본 발명의 또 다른 양태에 따르면, 본 발명은 우엉(Arctium lappa root) 추출물을 포함하는 우울증 개선 또는 예방용 식품 조성물을 제공한다.According to another aspect of the present invention, there is provided a food composition for improving or preventing depression comprising Arctium lappa root extract.
본 발명의 또 다른 양태에 따르면, 본 발명은 우엉(Arctium lappa root) 추출물을 포함하는 여성 갱년기 증후군 개선 또는 예방 식품 조성물을 제공한다.According to yet another aspect of the present invention, there is provided a composition for improving or preventing female menopausal syndrome comprising an extract of Arctium lappa root.
본 발명의 식품 조성물은 상술한 "우울증 예방 또는 치료용 약제학적 조성물"과 동일하게 유효성분으로서 우엉 추출물을 이용하기 때문에, 공통된 사항은 본 명세서의 과도한 중복성을 피하기 위해 그 기재를 생략한다.Since the food composition of the present invention uses the burdock extract as an active ingredient in the same manner as the above-mentioned " pharmaceutical composition for the prevention or treatment of depression ", the description thereof is omitted in order to avoid excessive redundancy in this specification.
본 발명의 조성물이 식품 조성물인 경우에는 분말, 과립, 정제, 캡슐 또는 음료 등의 형태로 제조될 수 있다. 예컨대 캔디류의 각종 식품류, 음료, 껌, 차, 비타민 복합제, 또는 건강보조 식품류 등이 있다.When the composition of the present invention is a food composition, it may be prepared in the form of powder, granule, tablet, capsule or beverage. For example, various foods of candy, beverages, gums, tea, vitamin complex, or health supplement foods.
본 발명의 식품 조성물은 유효성분으로서 우엉 추출물뿐만 아니라, 식품 제조 시에 통상적으로 첨가되는 성분을 포함할 수 있으며, 예를 들어, 단백질, 탄수화물, 지방, 영양소, 조미제 및 향미제를 포함한다. 상술한 탄수화물의 예는 모노사카라이드, 예를 들어, 포도당, 과당 등; 디사카라이드, 예를 들어 말토스, 슈크로스, 올리고당 등; 및 폴리사카라이드, 예를 들어 덱스트린, 사이클로덱스트린 등과 같은 통상적인 당 및 자일리톨, 소르비톨, 에리트리톨 등의 당알콜이다. 향미제로서 천연 향미제 [타우마틴, 스테비아 추출물 (예를 들어 레바우디오시드 A, 글리시르히진 등]) 및 합성 향미제(사카린, 아스파르탐 등)를 사용할 수 있다. 예컨대, 본 발명의 식품 조성물이 드링크제로 제조되는 경우에는 본 발명의 우엉 추출물 이외에 구연산, 액상과당, 설탕, 포도당, 초산, 사과산, 과즙, 두충 추출액, 대추 추출액, 감초 추출액 등을 추가로 포함시킬 수 있다.The food composition of the present invention may contain not only burdock extract as an active ingredient, but also components that are ordinarily added during the manufacture of a food, for example, proteins, carbohydrates, fats, nutrients, flavoring agents and flavoring agents. Examples of the above-mentioned carbohydrates are monosaccharides such as glucose, fructose, and the like; Disaccharides such as maltose, sucrose, oligosaccharides and the like; And polysaccharides such as dextrin, cyclodextrin and the like, and sugar alcohols such as xylitol, sorbitol and erythritol. Natural flavorings such as tau martin and stevia extract (e.g., rebaudioside A and glycyrrhizin) and synthetic flavorings (saccharine, aspartame, etc.) can be used as flavorings. For example, when the food composition of the present invention is prepared as a drink, it may further contain citric acid, liquid fructose, sugar, glucose, acetic acid, malic acid, juice, mulberry extract, jujube extract, licorice extract, have.
본 발명의 다른 양태에 따르면, 본 발명은 우엉(Arctium lappa root) 추출물을 포함하는 동물 우울증 또는 스트레스 개선 또는 예방 기능성 사료 첨가제 조성물을 제공한다.According to another aspect of the present invention, there is provided an animal depression or stress-relieving or preventive functional feed additive composition comprising an extract of Arctium lappa root.
본 발명의 또 다른 양태에 따르면, 본 발명은 우엉(Arctium lappa root) 추출물을 포함하는 동물 인지능력 향상 또는 개선 기능성 사료 첨가제 조성물을 제공한다.According to another aspect of the present invention, there is provided an animal cognitive ability enhancing or improving functional feed additive composition comprising an extract of Arctium lappa root.
우엉은 오랫동안 식용 및 민간약으로 사용되어 왔던 것이므로 이로부터 추출된 성분 또는 우엉 추출물 자체는 독성 및 부작용 등의 문제가 없을 것으로 예상할 수 있다. 이러한 이유로 우엉 추출물은 동일한 목적으로 동물의약품 및 기능성 사료로도 적용이 가능하다.Since burdock has been used for a long time as an edible and folk medicine, the components extracted from the burdock or the burdock extract itself may be expected to have no toxicity and side effects. For this reason, burdock extracts can be applied to animal drugs and functional feeds for the same purpose.
본 발명의 또 다른 양태에 따르면, 본 발명은 (a) 우엉(Arctium lappa root) 추출물의 치료학적 유효량 및 (b) 약제학적으로 허용되는 담체를 포함하는 인지기능 장애 예방 또는 치료용 약제학적 조성물을 제공한다.According to another aspect of the present invention, the present invention provides a pharmaceutical composition for the prevention or treatment of cognitive dysfunction comprising (a) a therapeutically effective amount of an extract of Arctium lappa root and (b) a pharmaceutically acceptable carrier, to provide.
본 발명의 또 다른 양태에 따르면, 본 발명은 우엉(Arctium lappa root) 추출물을 포함하는 인지능력 또는 기억력 개선 식품 조성물을 제공한다.In accordance with another aspect of the present invention, there is provided a cognitive or memory improving food composition comprising an extract of Arctium lappa root.
본 발명의 인지기능 장애 예방 또는 치료용 약제학적 조성물 및 인지능력 또는 기억력 개선 식품 조성물은 상술한 "우울증 예방 또는 치료용 약제학적 조성물"과 동일하게 유효성분으로서 우엉 추출물을 이용하기 때문에, 공통된 사항은 본 명세서의 과도한 중복성을 피하기 위해 그 기재를 생략한다.Since the pharmaceutical composition for cognitive dysfunction prevention or treatment and the composition for improving cognitive ability or memory of the present invention use burdock extract as an active ingredient in the same manner as the above-mentioned " pharmaceutical composition for preventing or treating depression & The description thereof is omitted in order to avoid excessive redundancy of the present specification.
본 발명의 일 구현예에 따르면, 상기 인지기능 장애는 치매, 학습장애, 실인증, 건망증, 실어증, 실행증 또는 섬망일 수 있다. 본 발명의 다른 구현예에 따르면, 상기 인지기능장애는 AIDS 유발 치매, 빈스완거 병, 루이소체 치매, 전두측두엽성 치매, 경도 인지 장애, 다발성 경색성 치매, 픽병, 의미치매, 알츠하이머성 치매 또는 혈관성 치매일 수 있다.According to an embodiment of the present invention, the cognitive dysfunction may be dementia, learning disability, impulsiveness, forgetfulness, aphasia, impression or delirium. According to another embodiment of the present invention, the cognitive dysfunction is selected from the group consisting of AIDS-induced dementia, Vince Werther's disease, duloxetine dementia, frontal temporal dementia, mild cognitive impairment, multiple infarct dementia, Pick disease, dementia of Alzheimer's, It can be dementia.
하기의 실시예에서 입증된 바와 같이, 우엉 추출물의 투여는 스트레스 또는 우울증 증상으로 인해 감소하는 BDNF의 함량을 정상수준으로 회복시켰다. 이와 같은 실험 데이터는 본 발명의 조성물이 우울증 또는 스트레스를 치료 또는 완화시키는데 이용될 수 있음을 보여주는 것이다.As demonstrated in the examples below, administration of Burdock Extracts restored the levels of BDNF to normal levels due to stress or depressive symptoms. Such experimental data show that the compositions of the present invention can be used to treat or alleviate depression or stress.
하기의 실시예에서 입증된 바와 같이, 우엉 추출물의 투여는 스트레스 또는 우울증 증상으로 인해 감소하는 BDNF의 함량을 정상수준으로 회복시켰다. BDNF는 뇌신경세포의 발달과 분화 및 생존에 필요한 요소로, 학습, 기억, 신경전달물질 발달에 중요한 역할을 나타낸다. 또한 우엉 추출물은 스트레스 부여에 따른 인지기능저하를 대조군대비 유의적으로 개선시켰으며, 양성대조군인 세이트존스워트(St. John's wort) 보다도 뛰어난 효능을 나타내었다. 결과적으로, 이와 같은 실험 데이터는 우엉 추출물이 스트레스 또는 우울증으로 인해 감소하는 학습 또는 인지기능을 개선시키데 이용될 수 있음을 보여주는 것이다.As demonstrated in the examples below, administration of Burdock Extracts restored the levels of BDNF to normal levels due to stress or depressive symptoms. BDNF is an essential factor in the development, differentiation and survival of neuronal cells, and plays an important role in learning, memory, and neurotransmitter development. In addition, burdock extract significantly improved the cognitive function depression by stress, compared with the control group, and showed better efficacy than the positive control group, St. John's wort. As a result, these experimental data show that burdock extract can be used to improve learning or cognitive function, which is reduced by stress or depression.
본 발명의 또 다른 양태에 따르면, 본 발명은 우엉(Arctium lappa root) 추출물을 포함하는 항스트레스 조성물을 제공한다.According to another aspect of the present invention, there is provided an anti-stress composition comprising an extract of Arctium lappa root.
본 발명의 항스트레스 조성물은 상술한 "우울증 예방 또는 치료용 약제학적 조성물"과 동일하게 유효성분으로서 우엉 추출물을 이용하기 때문에, 공통된 사항은 본 명세서의 과도한 중복성을 피하기 위해 그 기재를 생략한다.Since the anti-stress composition of the present invention uses the burdock extract as an active ingredient in the same manner as the above-mentioned " pharmaceutical composition for preventing or treating depression ", the description thereof is omitted in order to avoid excessive redundancy in this specification.
본 명세서에서 사용된 용어, "항스트레스"란 스트레스에 대한 저항 작용을 통칭하여 나타내는 용어로서, 예를들어, 스트레스의 예방 효과, 스트레스 감소 효과 또는 스트레스로 인한 다양한 증상(예컨대, 우울증, 인지기능 또는 학습 능력 감소, 두통, 소화불량, 피로, 피부의 거칠어짐, 신경증, 위궤양 등)의 억제 및 개선 효과를 나타내는 용어이다.As used herein, the term " antisstress " refers collectively to the resistance to stress, and includes, for example, a preventive effect of stress, a stress reduction effect or various symptoms due to stress A reduction in learning ability, headache, indigestion, fatigue, skin roughness, neurosis, gastric ulcer, etc.).
본 발명의 항스트레스 조성물은 약제학적 조성물 또는 식품 조성물로 제조될 수 있다.The anti-stress composition of the present invention may be manufactured from a pharmaceutical composition or a food composition.
본 발명의 특징 및 이점을 요약하면 다음과 같다:The features and advantages of the present invention are summarized as follows:
(a) 본 발명은 우엉 추출물을 유효성분으로 포함하는 우울증 예방 또는 치료용 약제학적 조성물; 여성 갱년기 증후군 예방 또는 치료용 약제학적 조성물; 기능성 사료 첨가제 조성물; 인지기능 장애 예방 또는 치료용 조성물; 또는 항스트레스 조성물에 관한 것이다.(a) a pharmaceutical composition for preventing or treating depression comprising burdock extract as an active ingredient; A pharmaceutical composition for the prevention or treatment of female menopausal syndrome; A functional feed additive composition; A composition for preventing or treating cognitive dysfunction; Or an anti-stress composition.
(b) 본 발명의 조성물은 신경세포 보호효과가 있으며, 특히 스트레스 또는 우울증 증상으로 인해 감소하는 BDNF (brain derivated neurotrophic factor) 발현 회복 효과 및 스트레스 부여에 따른 인지기능저하의 개선효과가 있다.(b) The composition of the present invention has a neuroprotective effect, and particularly has an effect of restoring the expression of brain derived neurotrophic factor (BDNF), which is decreased due to stress or depression, and the improvement of cognitive function by stress.
(c) 이로 인해, 종래 항우울제의 부작용을 극복하고 동시에 우울증 또는 스트레스 치료/개선효과 및 인지기능 향상 효과를 가져올 수 있다는 이점이 있다.(c) This has the advantage of overcoming side effects of conventional antidepressants and at the same time bringing about effects of depression or stress treatment / improvement and cognitive function improvement.
도 1은 산사 주정추출물의 신경세포 보호 효과를 나타내는 그래프이다. 덱사메타손을 첨가한 세포주(DEX)에서는 세포독성에 따라 유의적으로 통계적으로 (#, P<0.05) 약 68%의 생존율을 나타내었지만, 우엉 주정추출물을 첨가한 세포주에서는 최대 150% 이상의 생존율을 나타내었으며(100 μM), 50 μM 및 100 μM의 농도에서는 기존 항우울제인 이미프라민과 St. John's wort 처리한 세포주에 비해 확연한 세포 생존율을 나타내었다 (*, P<0.05; ***, P<0.001). 대조군에는 1% DMSO(dimethyl sulfoxide)용매를 처리하였다.
도 2는 우엉 주정추출물 마우스에 주입 후 강제 수영 실험을 실시한 결과를 나타낸 그래프이다. 우엉 추출물을 경구 투여한 그룹은 2차 증류수를 경구 투여한 그룹(Control)에 비하여 통계적으로 움직임 없이 가만히 있는 시간 비중(immobility time (%))이 유의적으로 감소하였다 (*, P<0.05; **, P<0.01).
도 3은 우엉 주정추출물이 스트레스 또는 우울증 증상으로 인해 브레인 영역에서 감소하는 뇌유래신경영양인자(BDNF)의 함량을 정상상태의 BDNF 수준으로 개선시키는 것으로 나타났다.
도 4는 우엉 추출물이 장시간 감금스트레스에 따라 감소되는 마우스의 인지기능을 개선시키는 결과로, 우엉 주정추출물은 스트레스부여에 따른 인지기능저하를 대조군대비 유의적으로 개선시켰다.FIG. 1 is a graph showing the protective effect of the extract of Sansa sp. The survival rate was about 68% in the dexamethasone-added cell line (DEX) according to the cytotoxicity (#, P <0.05) but the survival rate of the cell line supplemented with the burdock extract was 150% (100 μM), 50 μM and 100 μM, respectively. (*, P <0.05; ***, P <0.001) compared with the cell line treated with John's wort. The control group was treated with 1% DMSO (dimethyl sulfoxide) solvent.
FIG. 2 is a graph showing the result of forced swimming experiment after injection into burdock extract extract mice. The group of oral administration of burdock extract significantly decreased the immobility time (%) (*, P <0.05; *) without statistically moving compared with the control group administered with the second distilled water *, P < 0.01).
FIG. 3 shows that burdock alcohol extract improves the content of brain-derived neurotrophic factor (BDNF), which decreases in the brain region due to stress or depression, to a normal level of BDNF.
FIG. 4 shows that the burdock extract extract significantly improved the cognitive function of the mice when the stress was given to the mice.
이하, 실시예를 통하여 본 발명을 더욱 상세히 설명하고자 한다. 이들 실시예는 오로지 본 발명을 보다 구체적으로 설명하기 위한 것으로, 본 발명의 요지에 따라 본 발명의 범위가 이들 실시예에 의해 제한되지 않는다는 것은 당업계에서 통상의 지식을 가진 자에 있어서 자명할 것이다.Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood by those skilled in the art that these embodiments are only for describing the present invention in more detail and that the scope of the present invention is not limited by these embodiments in accordance with the gist of the present invention .
실시예Example
실험재료 및 실험방법Materials and Experiments
우엉 추출물Burdock extract
실험에 사용된 우엉(Arctium lappa L.)는 녹원에서 구입한 중국산으로, 우엉 600 g을 10배수의 주정 6 L에 투입한 후 진공저온 추출장치(COSMOS-660)를 사용하여 80 ℃에서 6시간 동안 추출하였다. 추출된 농축액을 동결 건조하여 우엉 추출분말을 얻었으며, 추출 수율은 17.2% 이었다.The burdock (Arctium lappa L.) used in the experiment was purchased from Kangwon, China. 600 g of burdock was added to 6 L of 10-fold alcohol, and then the mixture was heated at 80 ° C for 6 hours using a vacuum low-temperature extraction apparatus (COSMOS- Lt; / RTI > The extracted concentrate was lyophilized to obtain an extract powder of burdock, and the extraction yield was 17.2%.
세포배양Cell culture
우엉 추출물의 신경세포 보호 효과를 측정하기 위해 인간 신경아세포종(human neuroblastoma) 세포주인 SH-SY5Y을 ATCC(Manassas, VA, USA)에 분양받아 사용하였으며, 10% FBS (Gibco, Carlsbad, CA, USA) 및 1% 페니실린 스트렙토마이신을 첨가한 Dulbecco's modified Eagle medium (Gibco) 배지를 사용하여 37℃, 5% CO2 조건에서 배양하였으며, 2-3 일 간격으로 계대 배양하여 사용하였다.SH-SY5Y, a human neuroblastoma cell line, was distributed to ATCC (Manassas, Va., USA) in 10% FBS (Gibco, Carlsbad, CA, USA) And 5% CO 2 in Dulbecco's modified Eagle medium (Gibco) supplemented with 1% penicillin streptomycin. The cells were subcultured at 2-3 days intervals.
동물실험Animal experiment
동물 행동 실험을 통한 항우울증 효능평가를 위하여 무게가 25 g 내지 30 g의 수컷 ICR 마우스 (5주령, 코아텍, 경기도, 한국)를 제어온도 23±1 ℃에서 케이지당 5마리씩 보관하여 12시간 빛/어두움 사이클을 수행하였다. 상기 마우스를 1주일간 적응시켰다. 동물의 프로토콜은 한국식품연구원의 기관 동물 관리 및 사용위원회(IACUC)에 의해 승인되었다.For the evaluation of antidepressant efficacy by animal behavior test, male ICR mice weighing 25 g to 30 g (5 weeks old, Koatech, Gyeonggi-do, Korea) were kept at a controlled temperature of 23 ± 1 ℃ for 5 animals per cage, / Dark cycle. The mice were adapted for one week. The animal protocol was approved by the Korea Food Research Institute's Institutional Animal Care and Use Committee (IACUC).
시험예 1. 뇌신경세포 보호 효과 측정Test Example 1. Measurement of protective effect of brain nerve cell
배양된 SH-SY5Y 세포는 96-웰 플레이트에 2.5Х104 cells/well 농도로 분주하여 1일 동안 배양 후, 200 μM 덱사메타손(dexamethasone)을 첨가하여 뇌신경세포독성을 유발시켰고, 우엉 추출물을 농도별(10 μM, 50 μM 및 100 μM)로 처리한 뒤 48시간 더 배양하였다. 세포 생존율은 MTT 분석방법을 통해 확인하였다. 즉, pMTT(3-[4,5-dimethyl-thiazol-2-yl]-2,5-diphenyltetrazolium bromide, Sigma Co., U.S.A) 용액(0.3 mg/mL. final conc.)을 웰 당 20 μL씩 적용시킨 후 4시간 동안 배양하고, 배양액을 제거한 후 DMSO를 150 μL씩 넣어 5분간 방치하면서 MTT formazan을 용해하였으며, 5분후 ELISA 리더로 흡광도(450 nm, 25℃)를 측정하여, 대조군의 생존율에 대한 백분율로 나타내었다.Cultured SH-SY5Y cells were seeded at a concentration of 2.5 × 10 4 cells / well in a 96-well plate, cultured for 1 day, and then 200 μM dexamethasone was added to induce neuronal cytotoxicity. 10 [mu] M, 50 [mu] M and 100 [mu] M) for 48 hours. Cell viability was confirmed by MTT assay. That is, a solution (0.3 mg / mL final conc.) Of pMTT (3- [4,5-dimethyl-thiazol-2-yl] -2,5- diphenyltetrazolium bromide, Sigma Co., USA) MTT formazan was dissolved in 150 μL of DMSO for 5 min. After 5 min, the absorbance (450 nm, 25 ° C) was measured with an ELISA reader to determine the survival rate of the control group As a percentage.
도 1은 우엉 추출물의 뇌신경세포 보호 효과를 나타내는 그래프로, 덱사메타손을 첨가한 세포주(DEX)에서는 세포독성에 따라 약 58%의 생존율을 나타내었지만, 우엉 추출물을 첨가한 세포주에서는 최대 180% 이상의 생존율을 나타내었으며, 기존 항우울제인 이미프라민(imipramine)과 St. John's wort 처리한 세포주에 비해 확연히 높은 세포 생존율을 나타내었음을 확인하였다.FIG. 1 is a graph showing the protective effect of brain tumors on burdock extract. In the dexamethasone-added cell line (DEX), the survival rate was about 58% depending on cytotoxicity. And the existing antidepressants imipramine and imipramine. Cell survival rate was significantly higher than that of the cell line treated with John's wort.
시험예 2. 부동 시간 측정Test example 2. Floating time measurement
강제수영 행동실험은 가장 많이 사용되고 있는 우울증 동물모델로, 투명한 아크릴 원통(높이 20 cm, 지름 12 cm)에 22-24 ℃ 온도의 물을 10 cm 높이로 채워, 마우스를 강제수영 시키는 행동실험이다. 이 때 나타나는 마우스의 부동자세는 물속에서 빠져 나올 수 없는 환경에서 포기할 때 나타나는 행동으로, 부동자세 시간이 증가함에 따라 마우스의 상태는 사람의 우울증과 매우 유사한 생리학적 변화가 나타나게 된다. 기존의 다양한 약리작용의 항우울제는 이러한 부동자세 시간을 억제시킨다.The forced swimming behavior test is the most commonly used depressive animal model, in which a transparent acrylic cylinder (20 cm in height, 12 cm in diameter) is filled with water at 22-24 ° C at a height of 10 cm and forced swimming of the mouse. In this case, the immobility of the mouse is the behavior that occurs when it is given up in an environment where it can not escape from the water. As the immobility time increases, the state of the mouse becomes physiologically very similar to that of human depression. The existing antidepressants of various pharmacological actions suppress this immobility time.
강제수영 행동실험에 따른 부동식간과 수영시간을 측정하기 위해 우엉 추출물을 2차 증류수에 용해시킨 농도 300 mg/kg (0.1 mL/10 g무게)의 시료를 마우스에 경구 투여하고, 60분 후 행동실험을 시행하였다.In order to measure the swimming time and swimming time according to the forced swimming behavior test, mice were orally administered a sample of 300 mg / kg (0.1 mL / 10 g weight) in which burdock extract was dissolved in the second distilled water, .
강제 수영 행동 실험(FST)을 측정하기 위해 높이 30 cm 이상의 투명 비커에 15 cm만큼 물(온도 23-25 ℃)로 채우고 마우스를 입수시켜 6분 동안 강제수영 시켰다. 입수시켜 6분 동안 디지털 비디오파일로 녹화하여 행동분석 프로그램(SMART v3.0, Panlab SL, Barcelona, Spain)을 사용하여 부동 및 수영 시간을 분석하였다. 각각의 결과는 처음 2분간을 제외한 나머지 4분 동안을 기준으로 마우스가 수면위에 움직임 없이 가만히 있는 부동 시간(immobility time)과 움직이는 시간(swimming time)을 각각 분석하였다.In order to measure the forced swimming behavior test (FST), a transparent beaker with a height of 30 cm or more was filled with 15 cm of water (temperature 23-25 ° C) and the mice were swallowed for 6 minutes. Recorded for 6 minutes in a digital video file and analyzed for immobility and swimming time using a behavior analysis program (SMART v3.0, Panlab SL, Barcelona, Spain). Each result was analyzed for immobility time and swimming time while the mouse was not moving on the water surface for 4 minutes except for the first 2 minutes.
도 2는 우엉 추출물을 마우스에 주입 후 강제 수영 실험을 실시한 결과를 나타낸 그래프로, 우엉 추출물을 경구 투여한 그룹은 2차 증류수를 경구 투여한 대조군 그룹(Con)에 비하여 움직임 없이 가만히 있는 시간 비중(immobility time (%))이 유의적으로 감소하였음을 확인하였다. 우엉 투여군은 Hypericum perforatum (Saint Jone's Wort) 투여군 보다도 개선된 효과를 나타내었다. 즉, 우엉 추출물이 항우울 효능을 나타내는 것을 알 수 있었다.FIG. 2 is a graph showing the results of a forced swimming experiment after injecting a burdock extract into a mouse. In the group administered orally administered with Burdock extract, the time-specific gravity immobility time (%)) was significantly decreased. The burdock treated group showed more effective than Hypericum perforatum (Saint Jone's Wort) treated group. In other words, burdock extract showed antidepressant efficacy.
시험예 3. 뇌유래신경영양인자 (BDNF, brain derivated neurotrophic factor) 측정Test Example 3. Measurement of brain derived neurotrophic factor (BDNF)
강제 수영 실험을 실시하고 실험동물을 흡입 마취시킨 다음 복대정맥으로부터 채혈하고 방혈하여 실험동물을 희생하였다. 이후 뇌를 신속하게 적출한 다음 전두피질(frontal cortex)을 분리하여 액체질소에 넣어 동결시키고, -80℃ 초저온냉장고(deepfreezer)에 넣어 보관하였다. 동결 보관된 뇌 조직에 프로테아제 억제제 (Roche, Mannheim, Germany)가 첨가된 RIPA 버퍼(Sigma, Missouri, USA)를 넣고 호모게나이저(homogenizer)로 뇌 조직을 균질화 시킨 후, 4℃, 3000 rpm에서 15분간 원심분리하여 상등액을 취해 단백질을 분리하였다. 분리된 단백질의 농도를 BCA 단백질 분석키트 (Thermo scientific, Illinois, USA)를 이용하여 정량한 뒤 30 μg의 단백질을 100℃에서 5분간 가열한 후 10% 젤(Bio-Rad, California, USA)을 이용하여 80 V에서 1시간 30분 동안 전기영동 시켰다. 전기영동 후 젤에 분리된 단백질의 BDNF 함량을 "WesternSureTM PREMIUM Chemiluminescent Substrate"(LICOR, Nebraska, USA)를 사용하여 1분간 반응시킨 후 C-DiGit 기기의 image studio software(LI-COR, Nebraska, USA)를 사용하여 현상하였다.Experimental animals were sacrificed by drawing blood from the abdominal vein and bleeding. After the brain was rapidly removed, the frontal cortex was isolated, frozen in liquid nitrogen and stored in a -80 ° C deep freezer. RIPA buffer (Sigma, Missouri, USA) supplemented with protease inhibitor (Roche, Mannheim, Germany) was added to the frozen storage brain tissue and the brain tissue was homogenized with a homogenizer. The supernatant was separated by centrifugation for minute to separate proteins. 30 μg of protein was heated at 100 ° C for 5 minutes, and then 10% gel (Bio-Rad, California, USA) was added thereto. The protein concentration was measured using a BCA protein assay kit (Thermo scientific, And electrophoresed at 80 V for 1 hour and 30 minutes. After the electrophoresis, the BDNF content of the separated proteins in the gel was reacted for 1 minute using "WesternSure ™ PREMIUM Chemiluminescent Substrate" (LICOR, Nebraska, USA) and the image studio software (LI-COR, Nebraska, USA) Lt; / RTI >
도 3은 우엉 추출물이 스트레스 또는 우울증 증상으로 인해 감소하는 BDNF의 함량을 정상상태의 BDNF 수준으로 개선시키는 것으로 나타난다. BDNF는 뇌신경세포의 발달과 분화 및 생존에 필요한 요소로, 학습, 기억, 신경전달물질 발달에 중요한 역할을 나타낸다. 즉, 우엉 추출물이 스트레스 또는 우울증으로 인해 감소하는 학습 또는 인지기능을 개선시키는 것으로 판단할 수 있다.Figure 3 shows that Burdock Extract improves the amount of BDNF that is reduced due to stress or depressive symptoms to a steady state BDNF level. BDNF is an essential factor in the development, differentiation and survival of neuronal cells, and plays an important role in learning, memory, and neurotransmitter development. In other words, it can be judged that burdock extract improves learning or cognitive function which is reduced by stress or depression.
시험예 4. 수동회피실험 (passive avoidance test)Test Example 4. Passive avoidance test
우엉 추출물의 인지기능개선 효능을 측정하기 위하여 passive avoidance 장치(GEMINI avoidance system, SD instrument, CA, USA)를 사용하였다. 중간에 guillotine door를 사이로 두고 양쪽에 24 ㎝ Х 20 ㎝ Х 20 ㎝의 챔버가 있으며, 각 챔버의 바닥은 grid가 설치되어 있어 어두운 방에 전류가 흐르도록 설정하였다. 실험동물을 오른쪽 챔버에 넣으면 10초 동안의 적응시간 후 챔버에 불이 들어오며, 동시에 guillotine door가 열린다. 이때 마우스의 어두운 부분으로 움직이는 본능으로 인하여 어두운 곳으로 옮겨가는데, 마우스가 어두운 챔버로 들어가는 시간(step through latency)을 측정하여 평가하는 방법이다. 마우스가 어두운 방에 들어가는 순간 guillotine door 닫히며, 바닥의 grid에 0.5 ㎃의 전류를 2초간 흐르게 하여 어두운 방으로 들어간 마우스에게 전기충격을 주었다. 마우스를 꺼내어 케이지로 이동하고 24시간이 지난 후 다시 밝은 방에 두고 guillotine door을 개방하였을 때, 전날의 전기쇼크를 기억하여 상자의 검은 부분으로 들어가지 않는 시간을 측정하여 인지기능을 평가하였다.A passive avoidance system (GEMINI avoidance system, SD instrument, CA, USA) was used to measure the cognitive function of burdock extract. There is a chamber of 24 ㎝ 20 ㎝ 20 ㎝ on both sides with a guillotine door in the middle, and the bottom of each chamber is set up so that a grid is installed and a current flows in a dark room. When the animal is placed in the right chamber, the chamber is lit after 10 seconds of adaptation time and at the same time the guillotine door is opened. In this case, moving to the dark place due to the instinct to move to the dark part of the mouse is a method of measuring and measuring the step through latency of the mouse into the dark chamber. When the mouse entered the dark room, the guillotine door was closed, and a current of 0.5 mA was applied to the bottom grid for 2 seconds to give electric shock to the dark room. When the mouse was removed and moved to the cage and the guillotine door was opened again in a bright room after 24 hours, the cognitive function was evaluated by measuring the time when the electric shock of the previous day was remembered and it did not enter the black part of the box.
도 4는 우엉 추출물이 1일 1회 6시간 3주간 감금스트레스에 따라 감소되는 마우스의 인지기능을 개선시키는 결과이다. 장시간 감금스트레스를 부여받은 대조군은 감금 스트레스를 부여받지 않은 정상군에 비해 유의적으로 인지기능손상에 따른 행동을 나타내었지만, 우엉 추출물은 스트레스 부여에 따른 인지기능저하를 대조군대비 유의적으로 개선시켰으며, 양성대조군인 세이트존스워트(St. John's wort) 보다도 뛰어난 효능을 나타내었다. 따라서 우엉 추출물이 만성 스트레스 또는 우울증으로 인해 감소하는 학습 또는 인지기능을 개선시키는 것으로 판단할 수 있다.FIG. 4 shows the results of improving the cognitive function of mice in which burdock extract is reduced with resting stress for 6 hours and 3 weeks once a day. The control group, which had been given the long - term restraint stress, showed significant cognitive impairment behavior compared to the control group without restraint stress, but the burdock extract significantly improved the cognitive function depression , And was superior to the positive control group, St. John's wort. Therefore, it can be judged that burdock extract improves learning or cognitive function which is reduced by chronic stress or depression.
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