KR20120130973A - Pharmaceutical composition and functional food having anti-depressant activity, and preparation method of the same - Google Patents
Pharmaceutical composition and functional food having anti-depressant activity, and preparation method of the same Download PDFInfo
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- KR20120130973A KR20120130973A KR1020110049101A KR20110049101A KR20120130973A KR 20120130973 A KR20120130973 A KR 20120130973A KR 1020110049101 A KR1020110049101 A KR 1020110049101A KR 20110049101 A KR20110049101 A KR 20110049101A KR 20120130973 A KR20120130973 A KR 20120130973A
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- extract
- preventing
- licorice
- hyangbuja
- treating depression
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Abstract
Description
본 발명은 향부자, 백복령, 감초 및 황련의 추출물을 포함하는 우울증 예방 및 치료용 약학 조성물 및 건강기능식품, 그의 제조방법에 관한 것이다.
The present invention relates to a pharmaceutical composition for preventing and treating depression, and a dietary supplement, comprising the extracts of Hyangbuja, Paekbokyeong, licorice, and yellow lotus.
"현대병"이라 불리는 우울증은 "마음의 감기"라고도 불릴 정도로 많은 사람에게서 나타나며 오늘날 손꼽히는 고빈도 질환의 하나로 자리매김되고 있다. 심신을 동시에 악화시키는 광범위한 질환으로 치료받지 않으면 몇 개월에서 몇 년 동안 지속될 수 있으며 관계의 와해나 직업적인 생산성의 상실, 무능이나 죽음에 이를 수도 있어 개인뿐 아니라 가족, 사회에 큰 부담을 주는 질환이다. 주요 우울증성 장애의 평생 유병률은 여자의 경우 10-25%, 남자의 경우 5-12% 정도로, 여자가 남성에 비해 약 2배 이상 많다. 우리나라 우울증 환자 수는 '04년 54만1000명에서 '07년 67만2545명으로 24.3% 증가하였고 '07년 우울증으로 진료를 받은 환자 중에서 외래환자 수는 66만7188명으로 전체 환자의 99%를 차지해 외래환자의 구성이 월등히 높은 경향을 보였다.Depression, called "modern disease," occurs in so many people, also called "colds of heart," and has become one of the most frequent diseases of today. If not treated with a wide range of conditions that simultaneously exacerbate the mind and body, it can last for months or years and can lead to a breakdown in relationships, loss of professional productivity, incompetence or death, putting a strain on individuals, families and society. . The lifetime prevalence of major depressive disorders is 10-25% in women and 5-12% in men, and women are about twice as likely as men. The number of depressive patients in Korea increased by 24.3% from 541,000 in 2004 to 67,2545 in 2007. Out of patients treated for depression in 2007, the number of outpatients was 66,7188, 99% of all patients. The composition of outpatients was very high.
현재 임상에 사용되는 항우울제는 모두 모노아민계 신경전달물질의 작용을 강화하는 약물들인데, 신경세포의 세로토닌 재흡수, 노르에피네프린의 재흡수 등을 억제하는 물질 (SSRI, SNRI, TCAs 계열 등), 모노아민 산화효소 억제제(monoamine oxidase inhibitors)와 같이 세로토닌이나 도파민 등의 분해를 억제하는 물질 등이 그것이다. Antidepressants used in clinical practice are all drugs that enhance the action of monoamine-based neurotransmitters, and substances that inhibit serotonin reuptake of neurons and norepinephrine reuptake (SSRI, SNRI, TCAs series, etc.), Substances that inhibit the degradation of serotonin or dopamine, such as monoamine oxidase inhibitors.
하지만, 현재 우리나라의 우울증 환자 중 약 10-25%만이 항우울제 치료를 받고 있으며, 이중 40% 가량은 부작용 등으로 인해 순응도가 떨어져 치료를 중단하는 것으로 알려져 있고, 나아가 항우울제를 복용하더라도 반응이 없는 비반응군(non-responder)이 33%에 달하는 것으로 보고되고 있다. However, only 10-25% of depressed patients in Korea are currently receiving antidepressant treatment, and about 40% of them are known to stop treatment due to side effects, and even stop responding even when taking antidepressants. 33% of non-responders are reported.
따라서, 수천년 임상경험이 축척된 한의약에서 부작용과 경제성 및 유효성을 만족시키는 항우울제 개발에 대한 노력이 절실히 요구되는 실정이다. 상기 우울증 치료과 관련한 한방 조성물과 관련하여, 한국등록특허 제10-0881368호는 산약, 용안육, 감초, 대추, 소백 및 산조인을 복합추출물 제시하고 있으며, 한국공개특허 제10-2008-0106514호는 연교, 후박, 생강즙, 반하, 지각, 구멍버섯, 자소엽 및 감초의 혼합추출물을 기재하고 있을 뿐이다.
Therefore, there is an urgent need for the development of antidepressant drugs that satisfy side effects, economic feasibility, and efficacy in oriental medicine, which has accumulated thousands of years of clinical experience. In relation to the herbal composition related to the treatment of depression, Korean Patent No. 10-0881368 proposes a complex extract of Chinese medicine, longan meat, licorice, jujube, Sobaek and Sanjoin, and Korean Patent Publication No. 10-2008-0106514 discloses Yeonkyo, It merely describes mixed extracts of grommets, ginger juice, halves, crusts, puncture mushrooms, cotyledon and licorice.
본 발명은 생약 한약재인 향부자, 백복령, 감초 및 황련의 추출물을 포함하는 우울증 예방 및 치료용 약학 조성물 및 건강기능식품과 그의 제조방법을 통해, 우수한 항우울 효과를 가지면서도 부작용을 감소시키고 경제성과 유효성을 갖춘 항우울증 약학 조성물 및 건강기능식품을 제공하려는 것이다.
The present invention through the pharmaceutical composition for preventing and treating depression and dietary supplements, including the extracts of herbal medicine, hyangbuja, baekbokyeong, licorice and rhubarb, which are herbal medicines, and functional foods, and a method for producing the same, while reducing side effects and reducing economic and efficacy To provide an antidepressant pharmaceutical composition and health functional food.
상기 목적을 달성하기 위하여 본 발명의 실시예에 따르면, 향부자, 백복령, 감초 및 황련의 추출물을 포함하는 우울증 예방 및 치료용 약학조성물을 제공한다. According to an embodiment of the present invention to achieve the above object, it provides a pharmaceutical composition for the prevention and treatment of depression, including extracts of Hyangbuja, Paekbokyeong, licorice and sulfur.
본 발명에 따른 우울증 예방 및 치료용 약학조성물은, 향부자, 백복령, 감초 및 황련의 추출물의 향부자 : 백복령 : 감초 : 황련의 중량비율이 2 내지 6 : 1 내지 2 : 1 내지 2 : 1 내지 2 인 것을 특징으로 한다. The pharmaceutical composition for preventing and treating depression according to the present invention is a hyangbuja, baekbokyeong, licorice, and the extract of the hyangbuja: Baekbokyeong: licorice: the weight ratio of nasturtium is 2 to 6: 1 to 2: 1 to 2: 1: 1-2 It is characterized by.
본 발명에 따른 우울증 예방 및 치료용 약학조성물은, 향부자, 백복령, 감초 및 황련의 추출물의 향부자 : 백복령 : 감초 : 황련의 중량비율이 4 : 1 : 1 : 1 인 것을 특징으로 한다. The pharmaceutical composition for preventing and treating depression according to the present invention is characterized in that the weight ratio of Hyangbuja, Baekbokryeong, Licorice and Huanglian extract: Baekbokyeong: Licorice: Huanglian is 4: 1: 1: 1.
본 발명에 따른 우울증 예방 및 치료용 약학조성물은, 정제수를 포함한 물, 탄소수 1 내지 탄소수 4의 저급 알코올 또는 이들의 혼합용매로부터 선택된 용매로 가용한 추출물인 것을 특징으로 한다.The pharmaceutical composition for preventing and treating depression according to the present invention is characterized in that the extract is available as a solvent selected from water including purified water, a lower alcohol having 1 to 4 carbon atoms or a mixed solvent thereof.
본 발명에 따른 우울증 예방 및 치료용 약학조성물은, 과립제, 레모네이드제, 산제, 시럽제, 액제, 엑스제, 엘릭서제, 유동엑스제, 유제, 현탁제, 전제, 침제, 정제, 주정제, 캡슐제, 틴크제 및 환제로 이루어진 군에서 선택된 어느 하나인 제형인 것을 특징으로 한다. Pharmaceutical composition for preventing and treating depression according to the present invention, granules, lemonades, powders, syrups, solutions, extracts, elixirs, liquid extracts, emulsions, suspensions, premises, acupuncture, tablets, spirits, capsules It is characterized in that the formulation is any one selected from the group consisting of tin and pills.
본 발명에 따른 우울증 예방 및 치료용 약학조성물 제조방법은, 향부자, 백복령, 감초 및 황련에 용매를 가하고 추출하여 추출물을 수득하는 단계; 및 상기 추출물을 농축하는 단계를 포함한다.Method for producing a pharmaceutical composition for preventing and treating depression in accordance with the present invention comprises the steps of obtaining a extract by adding and extracting a solvent to Hyangbuja, Paekbokyeong, licorice and yellow lotus; And concentrating the extract.
본 발명에 따른 우울증 예방 및 치료용 약학조성물 제조방법은, 상기 추출물을 수득하는 단계는 향부자, 백복령, 감초 및 황련의 총 중량대비 5 내지 20배 중량의 용매를 가하여 추출하는 것을 특징으로 한다. Method for producing a pharmaceutical composition for preventing and treating depression according to the present invention, the step of obtaining the extract is characterized in that the extract by adding a solvent of 5 to 20 times the weight of the total weight of Hyangbuja, Baekbokyeong, licorice and sulfur.
본 발명에 따른 우울증 예방 및 치료용 약학조성물 제조방법은, 상기 추출물을 수득하는 단계는 80℃ 내지 120℃에서 1시간 내지 10시간 동안 추출하는 것을 특징으로 한다. In the method for preparing a pharmaceutical composition for preventing and treating depression according to the present invention, the step of obtaining the extract is characterized in that the extraction for 1 to 10 hours at 80 ℃ to 120 ℃.
본 발명에 따른 우울증 예방 및 치료용 약학조성물 제조방법은, 상기 농축하는 단계는 상기 추출물을 여과하는 단계; 상기 여과된 추출물을 감압 농축하는 단계, 상기 농축된 추출물을 건조하는 단계; 상기 건조된 농축물을 분쇄하여 분말화 하는 단계를 포함하는 것을 특징으로 한다. Method for producing a pharmaceutical composition for preventing and treating depression in accordance with the present invention, wherein the concentration step is filtering the extract; Concentrating the filtered extract under reduced pressure, and drying the concentrated extract; It characterized in that it comprises the step of pulverizing the dried concentrate concentrate.
본 발명에 따른 우울증 예방 및 치료용 건강기능식품은, 향부자, 백복령, 감초 및 황련의 추출물을 포함한다.Health functional foods for preventing and treating depression in accordance with the present invention, include extracts of Hyangbuja, Baekbokyeong, licorice and yellow lotus.
본 발명에 따른 우울증 예방 및 치료용 건강기능식품은, 상기 향부자, 백복령, 감초 및 황련의 추출물은 향부자 : 백복령 : 감초 : 황련의 중량비율이 2 내지 6 : 1 내지 2 : 1 내지 2 : 1 내지 2 인 것을 특징으로 한다. Health functional foods for the prevention and treatment of depression according to the present invention, the extract of the Hyangbuja, Paekbokyeong, licorice and yellow lotus is a hyangbuja: Baekbokyeong: Licorice: Lactose weight ratio of 2 to 6: 1 to 2: 1 to 2: 1 to 2. It is characterized by two.
본 발명에 따른 우울증 예방 및 치료용 건강기능식품은, 상기 향부자, 백복령, 감초 및 황련의 추출물은 향부자 : 백복령 : 감초 : 황련의 중량비율이 4 : 1 : 1 : 1 인 것을 특징으로 한다. Health functional foods for the prevention and treatment of depression according to the present invention, the extract of Hyangbuja, Paekbokyeong, licorice and Huangren is characterized in that the weight ratio of Hyangbuja: Baekbokyeong: Licorice: Huanglian is 4: 1: 1: 1.
본 발명에 따른 우울증 예방 및 치료용 건강기능식품은, 상기 추출물은 정제수를 포함한 물, 탄소수 1 내지 4의 저급 알코올 또는 이들의 혼합용매로부터 선택된 용매로 가용한 추출물인 것을 특징으로 한다. Health functional foods for preventing and treating depression according to the present invention, the extract is characterized in that the extract is available as a solvent selected from water, including purified water, lower alcohol having 1 to 4 carbon atoms or a mixed solvent thereof.
본 발명에 따른 우울증 예방 및 치료용 건강기능식품은, 상기 건강기능식품은 캡슐, 정제, 분말, 과립, 액상, 환, 편상, 페이스트상, 시럽, 겔, 젤리 및 바로 이루어진 군에서 선택된 어느 하나의 제형인 것을 특징으로 한다. Health functional food for preventing and treating depression according to the present invention, the health functional food is any one selected from the group consisting of capsules, tablets, powders, granules, liquid, pills, flakes, paste, syrup, gel, jelly and bar It is characterized in that the formulation.
본 발명에 따른 우울증 예방 및 치료용 건강기능식품의 제조방법은, 향부자, 백복령, 감초 및 황련에 용매를 가하고 추출하여 추출물을 수득하는 단계; 및 상기 추출물을 농축하는 단계를 포함한다.Method for producing a health functional food for preventing and treating depression according to the present invention comprises the steps of obtaining a extract by adding and extracting a solvent to Hyangbuja, baekbokyeong, licorice and yellow lotus; And concentrating the extract.
본 발명에 따른 우울증 예방 및 치료용 건강기능식품의 제조방법은, 상기 추출물을 수득하는 단계는 향부자, 백복령, 감초 및 황련의 총 중량대비 5 내지 20배 중량의 용매를 가하여 추출하는 것을 특징으로 한다. Method for producing a health functional food for preventing and treating depression according to the present invention, the step of obtaining the extract is characterized in that the extract by adding a solvent of 5 to 20 times the weight of the total weight of Hyangbuja, Baekbokyeong, licorice and rhubarb .
본 발명에 따른 우울증 예방 및 치료용 건강기능식품의 제조방법은, 상기 추출물을 수득하는 단계는 80℃ 내지 120℃에서 1시간 내지 10시간 동안 추출하는 것을 특징으로 한다. Method for producing a health functional food for preventing and treating depression according to the present invention, the step of obtaining the extract is characterized in that the extraction for 1 to 10 hours at 80 ℃ to 120 ℃.
본 발명에 따른 우울증 예방 및 치료용 건강기능식품의 제조방법은, 상기 농축하는 단계는 상기 추출물을 여과하는 단계; 상기 여과된 추출물을 감압 농축하는 단계, 상기 농축된 추출물을 건조하는 단계; 상기 건조된 농축물을 분쇄하여 분말화 하는 단계를 포함하는 것을 특징으로 한다.
Method for producing a health functional food for preventing and treating depression in accordance with the present invention, the step of concentrating the step of filtering the extract; Concentrating the filtered extract under reduced pressure, and drying the concentrated extract; It characterized in that it comprises the step of pulverizing the dried concentrate concentrate.
본 발명에 의하면 우울증상 및 불안 증상을 현저하게 개선하고, 뇌 조직의 산화 스트레스나 신경 독성을 유의적으로 억제하여 현저하게 개선된 우울증 예방 및 치료효과를 갖는 우울증 약제 또는 건강기능식품을 제조할 수 있다.
According to the present invention can significantly reduce the depressive symptoms and anxiety symptoms, and significantly inhibit the oxidative stress or neurotoxicity of the brain tissue to prepare a depression drug or health functional food having a significantly improved depression prevention and treatment effect have.
도 1은 본 발명에 따른 일 실시예의 항우울효과를 검증하기 위한 실험설계모델이다.
도 2는 꼬리 매달기 실험(Tail suspension test) 결과이다.
도 3은 강제 수영 실험(Forced swimming test) 결과이다.
도 4는 새로운 환경내 섭식 제한 실험(Novelty-suppressed feeding) 결과이다.
도 5는 구슬 묻기 실험(Marble burying test) 결과이다.
도 6은 과산화수소 자극으로부터 세포 보호 효과를 확인하는 세포 생존실험결과이다.
도 7는 세포 내 활성 산소종을 형광분석하여 항산화력을 측정한 결과이다.
도 8은 마우스 해마의 말론디알데하이드(Malondialdehyde, MDA) 양을 측정한 결과이다.1 is an experimental design model for verifying the antidepressant effect of one embodiment according to the present invention.
2 is a tail suspension test result.
3 is a result of a forced swimming test.
Figure 4 shows the results of a novel environment-suppressed feeding experiment.
5 is a result of marble burying test.
6 is a cell survival test result confirming the protective effect from hydrogen peroxide stimulation.
Figure 7 is the result of measuring the antioxidant power by fluorescence analysis of intracellular reactive oxygen species.
8 is a result of measuring the amount of malondialdehyde (MDA) of mouse hippocampus.
상기 상술한 목적을 달성하기 위한 본 발명의 향부자, 백복령, 감초 및 황련의 추출물을 포함하는 우울증 예방 및 치료용 약학조성물 및 우울증 예방 및 치료용 건강기능식품과 그 제조방법에 특징이 있는 것을 구성으로 한다. 이하 첨부된 도면을 이용하여 본 발명의 구성을 더욱 상세하게 살펴보도록 한다.Depressive and therapeutic pharmaceutical composition for depression and anti-depressive and pharmaceutical composition comprising the extract of Hyangbuja, Baekbokyeong, Licorice and Hwangju of the present invention for achieving the above-mentioned object and characterized in that the manufacturing method do. Hereinafter, the configuration of the present invention will be described in detail with reference to the accompanying drawings.
상기 향부자(香附子)는 방동사니과(사초과, Cyperaceae)에 속한 다년생 초본인 향부자 Cyperus rotundus L. 의 뿌리이다. 성질은 평(平)하고, 맵고 달고 쓴 맛을 지니고, 기를 통하게 하고 기혈이 몰려 있는 것을 푸는 효능이 있다.Hyangbuja (香附 子) is the root of the perennial herb, Cyperus rotundus L., a perennial herb, belonging to the genus Cyperaceae. It has a flat, spicy, sweet and bitter taste, and it has the effect of letting the qi and bleeding.
백복령(白茯笭)은 구멍쟁이 버섯과에 속한 균류식물인 복령균 Poria cocas(Schw.) Wolf.의 건조한 균핵이다. 성분이 고르며 맛이 달며 독이 없고 위를 열어주고 구역을 멎게 하며 심신을 편하게 하고 비장을 보하고 가래를 삭이고 기억력 감퇴를 낫게 하는 등, 위, 간, 이자, 신장 등의 질병에도 탁월한 효과가 있는 것으로 알려져 있다. 약리실험 결과에 의하면 이뇨작용과 혈당량을 낮추는 작용 및 변역 부활작용을 하는 것으로 알려져 있다. Baekbokryeong (白 茯 笭) is a dry fungal nucleus of Poria cocas (Schw.) Wolf. It has an excellent effect on diseases such as stomach, liver, interest, kidney, etc., with even ingredients, sweet taste, no poison, open stomach, swelling area, relax mind and body, spleen, sputum, and memory loss. Known. According to pharmacological experiments, it is known that diuretic action and blood sugar lowering action and translational reactivation action.
황련(黃連)은 미나리아재비과 황련(Coptis chinensis, Coptis japonica)의 뿌리로 우리나라에서는 매자나무과의 깽깽이풀(Jeffersonia dubia)의 뿌리를 상황련(常黃連)이라 칭하며 대용한다. 성질은 차고, 맛은 쓰다. 열기를 식히고 습기를 제거하고, 심장의 열을 제거하는 효능을 갖는다.Huanglian (黄連) is the root of the buttercup and Coptis chinensis ( Coptis japonica), the root of the barberry of the barberry, Jeffersonia dubia in Korea, is called the situation lotus (常 黃連). The nature is cold, the taste is bitter. It has the effect of cooling the heat, removing moisture, and removing the heat of the heart.
감초(甘草)는 콩과에 속한 다년생 초본인 감초(Glycyrrhiza ssp.)의 뿌리와 근상경(根狀莖)이다. 따뜻하고 맛이 달며 독이 없고 백가지의 독을 풀어 주고 구토의 정이 되며 72가지의 석재와 1200가지의 초재를 고루 완화하고 모든 약으로 해서 효력이 나도록 하기 때문에 일명 국로라 한다. Licorice (甘草) is the root of the perennial herbaceous persimmon ( Glycyrrhiza ssp.) And the root of the root. It is called a souk because it is warm, sweet, without poison, releasing hundred poisons, giving vomiting, and alleviating 72 stones and 1200 herbaceous ingredients and making them effective with all medicines.
본 발명에 따른 우울증 예방 및 치료용 약학조성물은, 향부자, 백복령, 감초 및 황련의 추출물을 포함한다. The pharmaceutical composition for preventing and treating depression according to the present invention includes extracts of Hyangbuja, Baekbokryeong, licorice, and yellow lotus.
본 발명에 따른 우울증 예방 및 치료용 약학조성물은, 향부자, 백복령, 감초 및 황련의 추출물의 향부자 : 백복령 : 감초 : 황련의 중량비율이 2 내지 6 : 1 내지 2 : 1 내지 2 : 1 내지 2 인 것으로 하며, 더욱 바람직하게는 4 : 1 : 1 : 1 인 것을 특징으로 한다. 이와 같은 비율에 있어 항우울증 효과가 가장 우수한 것으로 나타났다.The pharmaceutical composition for preventing and treating depression according to the present invention is a hyangbuja, baekbokyeong, licorice, and the extract of the hyangbuja: Baekbokyeong: licorice: the weight ratio of nasturtium is 2 to 6: 1 to 2: 1 to 2: 1: 1-2 More preferably, it is characterized by being 4: 1: 1: 1. In this ratio, the antidepressant effect was found to be the best.
본 발명에 따른 우울증 예방 및 치료용 약학조성물의 추출에 사용되는 추출용매는 약학 조성물이나 생약제 추출시에 사용되는 용매이면 특정 용매에 한정되지 않는다. 보다 바람직한 추출물은 정제수를 포함한 물, 탄소수 1 내지 탄소수 4의 저급 알코올 또는 이들의 혼합용매로부터 선택된 용매로 가용한 추출물이다.The extraction solvent used for the extraction of the pharmaceutical composition for preventing and treating depression according to the present invention is not limited to a specific solvent as long as it is a solvent used for extracting the pharmaceutical composition or the herbal medicine. More preferred extract is an extract available in a solvent selected from water including purified water, lower alcohols having 1 to 4 carbon atoms or mixed solvents thereof.
본 발명에 따른 우울증 예방 및 치료용 약학조성물은, 그 제형에 있어 일반적으로 약학 조성물에 적용되는 제형이면 특정 제형에 제한되지 않고, 상기 추출물을 주원료로 하여 제형 구성에 일반적으로 사용되는 보조제나 첨가제 등을 더욱 함유하여 다양한 제형을 제공할 수 있다. 보다 바람직하게는 과립제, 레모네이드제, 산제, 시럽제, 액제, 엑스제, 엘릭서제, 유동엑스제, 유제, 현탁제, 전제, 침제, 정제, 주정제, 캡슐제, 틴크제 및 환제로 이루어진 군에서 선택된 어느 하나인 제형인 것을 특징으로 한다. The pharmaceutical composition for preventing and treating depression according to the present invention is not limited to a specific formulation as long as it is a formulation generally applied to a pharmaceutical composition in the formulation, and supplements or additives commonly used in the composition of the formulation using the extract as a main ingredient. It may further contain a variety of formulations. More preferably in the group consisting of granules, lemonades, powders, syrups, liquids, extracts, elixirs, liquid extracts, emulsions, suspensions, premises, acupuncture, tablets, spirits, capsules, tinctures and pills It is characterized in that the formulation is any one selected.
본 발명에 따른 우울증 예방 및 치료용 약학조성물 제조방법은, 향부자, 백복령, 감초 및 황련에 용매를 가하고 추출하여 추출물을 수득하는 단계; 및 상기 추출물을 농축하는 단계를 포함한다. Method for producing a pharmaceutical composition for preventing and treating depression in accordance with the present invention comprises the steps of obtaining a extract by adding and extracting a solvent to Hyangbuja, Paekbokyeong, licorice and yellow lotus; And concentrating the extract.
상기 향부자, 백복령, 감초 및 황련은 일반적으로 사용되는 건조물을 통째로 추출할 수도 있고, 적당한 크기로 잘라서 (예컨대 길이 3 내지 20 ㎝) 추출하거나, 또는 분쇄하거나 분말로 갈아서 추출할 수 있다. 일반적으로 가장 효과적으로 유효성분을 추출하기 위해서는 작게 부수어 추출할수록 바람직하다.The saengbuja, baekbokyeong, licorice, and yellow lotus may be extracted as a whole, dried materials generally used, and may be extracted by cutting to a suitable size (for example, 3 to 20 cm in length), or pulverized or ground into powder. In general, in order to extract the active ingredient most effectively, the smaller the extraction, the better.
상기 추출물을 수득하는 단계에서 향부자, 백복령, 감초 및 황련과 용매의 비율은 일반적으로 생약제 추출에 사용될 수 있는 정도의 범위 내에서 사용가능하며, 바람직하게는 향부자, 백복령, 감초 및 황련의 총 중량대비 5 내지 20배 중량의 용매를 가하여 추출하는 것을 특징으로 한다. 상기보다 용매의 양이 더욱 많은 경우에는 유효 성분의 농도가 낮거나 추출에 드는 에너지나 시간이 많이 소모되어 바람직하지 않고, 이보다 더 적은 경우에는 최종 추출물의 농도가 너무 높거나 수율이 감소하여 바람직하지 않을 수 있다.In the step of obtaining the extract, the ratio of Hyangbuja, Baibokyeong, licorice, and yellow lotus and the solvent are generally available within the range that can be used for extracting the herbal medicine, and preferably, compared to the total weight of Hyangbuja, Baibokyeong, Licorice and Huangyeon. It is characterized by extracting by adding a solvent of 5 to 20 times the weight. If the amount of the solvent is higher than the above, the concentration of the active ingredient is low or energy and time required for extraction are not preferable, and if it is less than this, the concentration of the final extract is too high or the yield is reduced, which is not preferable. You may not.
상기 추출물을 수득하는 단계는 통상적으로 생약재를 추출하는 조건이면 특별한 제한 없이 사용 가능하며, 바람직하게는 80℃ 내지 120℃에서 1시간 내지 10시간 동안 추출하는 것을 특징으로 한다. 그 중에서도 95℃ 내지 100℃에서 3시간 내지 5시간 동안 추출하는 것이 가장 효과적이다. 상기 온도보다 더 낮거나 시간이 짧은 경우에는 유효성분의 추출이 효과적이지 않고, 상기 온도보다 더 높거나 시간이 긴 경우에는 추출 공정의 시간이나 효율면에서 바람직하지 않고 또한 유효성분이 파괴되거나 바람직하지 않은 성분이 혼합되거나 맛이 변할 우려도 있다. Obtaining the extract may be used without any particular limitation as long as it is a condition for extracting the herbal medicine, and preferably characterized in that it is extracted for 1 hour to 10 hours at 80 ℃ to 120 ℃. Among them, it is most effective to extract for 3 hours to 5 hours at 95 ℃ to 100 ℃. If the temperature is lower or shorter than the temperature, the extraction of the active ingredient is not effective. If the temperature is higher or longer than the temperature, the extraction of the active ingredient is not preferable in terms of time or efficiency of the extraction process, and the active ingredient is destroyed or undesirable. The ingredients may be mixed or taste may change.
보다 구체적으로 상기 농축하는 단계는 보다 구체적으로 상기 추출물을 여과하는 단계; 상기 여과된 추출물을 감압 농축하는 단계, 상기 농축된 추출물을 건조하는 단계; 상기 건조된 농축물을 분쇄하여 분말화 하는 단계를 포함하는 것을 특징으로 한다. More specifically, the step of concentrating more specifically, filtering the extract; Concentrating the filtered extract under reduced pressure, and drying the concentrated extract; It characterized in that it comprises the step of pulverizing the dried concentrate concentrate.
상기 추출물을 여과, 농축, 건조, 분쇄하는 방법은 추출물에 대하여 통상적으로 적용할 수 있는 여과, 농축, 건조, 분쇄방법이면 특정한 방법에 국한되지 않는다. 보다 바람직하게는 여과과정에서는 추출된 유효성분이 여과액 속에 포함될 수 있으면서 불순물은 제거할 수 있는 적당한 크기의 필터를 사용하고, 여과에서 분쇄의 일련의 과정은 유효성분의 파괴를 막기 위하여 60℃이하에서 진행하는 것이 바람직하다. The method of filtering, concentrating, drying and pulverizing the extract is not limited to a specific method as long as it is a filtration, concentration, drying and pulverization method that can be commonly applied to the extract. More preferably, in the filtration process, a filter of a suitable size capable of removing impurities while containing the extracted active ingredient may be included in the filtrate, and a series of pulverization in filtration may be performed at 60 ° C. or lower to prevent destruction of the active ingredient. It is preferable to proceed.
또한 생약재 내 존재하는 유효성분의 충분한 추출을 위하며, 상기 과정에 의해 1회 추출한 생약재에 다시 한번 용매를 가하여 다시 한번 추출하는 과정을 부가할 수 있다. 이 경우 첫 번째 추출한 추출물과 두 번째 추출한 추출물을 혼합하여 동시에 여과 및 농축과정을 거친다면 더욱 효율적이고 바람직하다.In addition, for the sufficient extraction of the active ingredient present in the herbal medicine, by adding the solvent once again to the herbal medicine extracted once by the above process may be added to the extraction process again. In this case, it is more efficient and preferable if the first and second extracts are mixed and filtered and concentrated at the same time.
본 발명에 따른 우울증 예방 및 치료용 건강기능식품은, 향부자, 백복령, 감초 및 황련의 추출물을 포함한다. Health functional foods for preventing and treating depression in accordance with the present invention, include extracts of Hyangbuja, Baekbokyeong, licorice and yellow lotus.
본 발명에 따른 우울증 예방 및 치료용 건강기능식품은, 향부자, 백복령, 감초 및 황련의 추출물의 향부자 : 백복령 : 감초 : 황련의 중량비율이 2 내지 6 : 1 내지 2 : 1 내지 2 : 1 내지 2 인 것으로 하며, 더욱 바람직하게는 4 : 1 : 1 : 1 인 것을 특징으로 한다. 이와 같은 비율에 있어 항우울증 효과가 가장 우수한 것으로 나타났다.Health functional foods for the prevention and treatment of depression according to the present invention, hyangbuja, baekbokyeong, licorice and the extract of the hyangbuja baekbuyeong: Licorice: licorice: the weight ratio of the yellow lotus is 2 to 6: 1 to 2: 1 to 2: 1 to 2 It is characterized by being more preferably 4: 1: 1: 1. In this ratio, the antidepressant effect was found to be the best.
본 발명에 따른 우울증 예방 및 치료용 건강기능식품의 추출에 사용되는 추출용매는 건강기능식품이나 생약제 추출시에 사용되는 용매이면 특정 용매에 한정되지 않는다. 보다 바람직한 추출물은 정제수를 포함한 물, 탄소수 1 내지 탄소수 4의 저급 알코올 또는 이들의 혼합용매로부터 선택된 용매로 가용한 추출물이다.Extraction solvent used for the extraction of health functional food for preventing and treating depression according to the present invention is not limited to a specific solvent as long as it is a solvent used in the extraction of health functional food or herbal medicines. More preferred extract is an extract available in a solvent selected from water including purified water, lower alcohols having 1 to 4 carbon atoms or mixed solvents thereof.
본 발명에 따른 우울증 예방 및 치료용 건강기능식품은, 그 제형에 있어 일반적으로 건강기능식품에 적용되는 제형이면 특정 제형에 제한되지 않고, 상기 추출물을 주원료로 하여 제형 구성에 일반적으로 사용되는 보조제나 첨가제 등을 더욱 함유하여 다양한 제형을 제공할 수 있다. 보다 바람직하게는 캡슐, 정제, 분말, 과립, 액상, 환, 편상, 페이스트상, 시럽, 겔, 젤리 및 바로 이루어진 군에서 선택된 어느 하나의 제형인 것을 특징으로 한다. The functional food for preventing and treating depression according to the present invention is not limited to a specific dosage form as long as the dosage form is generally applied to the health functional food in the dosage form. It may further contain additives and the like to provide a variety of formulations. More preferably, it is characterized in that the formulation of any one selected from the group consisting of capsules, tablets, powders, granules, liquid, pill, flaky, paste, syrup, gel, jelly and bar.
본 발명에 따른 우울증 예방 및 치료용 건강기능식품 제조방법은, 향부자, 백복령, 감초 및 황련에 용매를 가하고 추출하여 추출물을 수득하는 단계; 및 상기 추출물을 농축하는 단계를 포함한다. Method for producing a health functional food for preventing and treating depression according to the present invention comprises the steps of obtaining a extract by adding and extracting a solvent to Hyangbuja, baekbokyeong, licorice and yellow lotus; And concentrating the extract.
상기 향부자, 백복령, 감초 및 황련은 일반적으로 사용되는 건조물을 통째로 추출할 수도 있고, 적당한 크기로 잘라서 (예컨대 길이 3 내지 20 ㎝) 추출하거나, 또는 분쇄하거나 분말로 갈아서 추출할 수 있다. 일반적으로 가장 효과적으로 유효성분을 추출하기 위해서는 작게 부수어 추출할수록 바람직하다.The saengbuja, baekbokyeong, licorice, and yellow lotus may be extracted as a whole, dried materials generally used, and may be extracted by cutting to a suitable size (for example, 3 to 20 cm in length), or pulverized or ground into powder. In general, in order to extract the active ingredient most effectively, the smaller the extraction, the better.
상기 추출물을 수득하는 단계에서 향부자, 백복령, 감초 및 황련과 용매의 비율은 일반적으로 생약제 추출에 사용될 수 있는 정도의 범위 내에서 사용가능하며, 바람직하게는 향부자, 백복령, 감초 및 황련의 총 중량대비 5 내지 20배 중량의 용매를 가하여 추출하는 것을 특징으로 한다. 상기보다 용매의 양이 더욱 많은 경우에는 유효 성분의 농도가 낮거나 추출에 드는 에너지나 시간이 많이 소모되어 바람직하지 않고, 이보다 더 적은 경우에는 최종 추출물의 농도가 너무 높거나 수율이 감소하여 바람직하지 않을 수 있다.In the step of obtaining the extract, the ratio of Hyangbuja, Baibokyeong, licorice, and yellow lotus and the solvent are generally available within the range that can be used for extracting the herbal medicine, and preferably, compared to the total weight of Hyangbuja, Baibokyeong, Licorice and Huangyeon. It is characterized by extracting by adding a solvent of 5 to 20 times the weight. If the amount of the solvent is higher than the above, the concentration of the active ingredient is low or energy and time required for extraction are not preferable, and if it is less than this, the concentration of the final extract is too high or the yield is reduced, which is not preferable. You may not.
상기 추출물을 수득하는 단계는 통상적으로 생약재를 추출하는 조건이면 특별한 제한 없이 사용 가능하며, 바람직하게는 80℃ 내지 120℃에서 1시간 내지 10시간 동안 추출하는 것을 특징으로 한다. 그 중에서도 95℃ 내지 100℃에서 3시간 내지 5시간 동안 추출하는 것이 가장 효과적이다. 상기 온도보다 더 낮거나 시간이 짧은 경우에는 유효성분의 추출이 효과적이지 않고, 상기 온도보다 더 높거나 시간이 긴 경우에는 추출 공정의 시간이나 효율면에서 바람직하지 않고 또한 유효성분이 파괴되거나 바람직하지 않은 성분이 혼합되거나 맛이 변할 우려도 있다. Obtaining the extract may be used without any particular limitation as long as it is a condition for extracting the herbal medicine, and preferably characterized in that it is extracted for 1 hour to 10 hours at 80 ℃ to 120 ℃. Among them, it is most effective to extract for 3 hours to 5 hours at 95 ℃ to 100 ℃. If the temperature is lower or shorter than the temperature, the extraction of the active ingredient is not effective. If the temperature is higher or longer than the temperature, the extraction of the active ingredient is not preferable in terms of time or efficiency of the extraction process, and the active ingredient is destroyed or undesirable. The ingredients may be mixed or taste may change.
보다 구체적으로 상기 농축하는 단계는 보다 구체적으로 상기 추출물을 여과하는 단계; 상기 여과된 추출물을 감압 농축하는 단계, 상기 농축된 추출물을 건조하는 단계; 상기 건조된 농축물을 분쇄하여 분말화 하는 단계를 포함하는 것을 특징으로 한다. More specifically, the step of concentrating more specifically, filtering the extract; Concentrating the filtered extract under reduced pressure, and drying the concentrated extract; It characterized in that it comprises the step of pulverizing the dried concentrate concentrate.
상기 추출물을 여과, 농축, 건조, 분쇄하는 방법은 추출물에 대하여 통상적으로 적용할 수 있는 여과, 농축, 건조, 분쇄방법이면 특정한 방법에 국한되지 않는다. 보다 바람직하게는 여과과정에서는 추출된 유효성분이 여과액 속에 포함될 수 있으면서 불순물은 제거할 수 있는 적당한 크기의 필터를 사용하고, 여과에서 분쇄의 일련의 과정은 유효성분의 파괴를 막기 위하여 60℃이하에서 진행하는 것이 바람직하다. The method of filtering, concentrating, drying and pulverizing the extract is not limited to a specific method as long as it is a filtration, concentration, drying and pulverization method that can be commonly applied to the extract. More preferably, in the filtration process, a filter of a suitable size capable of removing impurities while containing the extracted active ingredient may be included in the filtrate, and a series of pulverization in filtration may be performed at 60 ° C. or lower to prevent destruction of the active ingredient. It is preferable to proceed.
또한 생약재 내 존재하는 유효성분의 충분한 추출을 위하며, 상기 과정에 의해 1회 추출한 생약재에 다시 한번 용매를 가하여 다시 한번 추출하는 과정을 부가할 수 있다. 이 경우 첫 번째 추출한 추출물과 두 번째 추출한 추출물을 혼합하여 동시에 여과 및 농축과정을 거친다면 더욱 효율적이고 바람직하다.
In addition, for the sufficient extraction of the active ingredient present in the herbal medicine, by adding the solvent once again to the herbal medicine extracted once by the above process may be added to the extraction process again. In this case, it is more efficient and preferable if the first and second extracts are mixed and filtered and concentrated at the same time.
이하에서는 실시예를 통하여 본 발명을 더욱 상세히 설명하고자 한다. 다만, 이들 실시예는 오로지 본 발명을 예시하기 위한 것으로서, 본 발명의 범위가 이들 실시예에 의해 제한되는 것으로 해석되지는 않는다 할 것이다.
Hereinafter, the present invention will be described in more detail with reference to Examples. It should be understood, however, that these examples are for illustrative purposes only and are not to be construed as limiting the scope of the present invention.
본 발명에 따른 실시예 1은 다음과 같은 과정을 통해 제조하였다. Example 1 according to the present invention was prepared through the following process.
향부자, 백복령, 감초 및 황련를 시중에서 구입하여 향부자 128g, 백복령 32g, 감초 32g 및 황련 32g을 파쇄 후 2 mesh 체를 통과하여 거른다. 상기 분말을 추출기에 넣은 다음 정제수를 1.8 L 가하고, 95 내지 100℃ 에서 3시간 가온 추출한다. 상기 추출물을 25㎛ 필터를 사용하여 여과하고 여액을 농축기를 이용하여 50 내지 60℃ 에서 감압 농축한다. 상기 한약 잔류물에 상기와 동일한 방법으로 정제수를 가하고 가온추출하는 과정을 1회 반복하여 농축액을 전부 합한다. 상기 합한 농축액을 건조기에 넣고 60℃ 이하에서 건조한다. 상기 수득한 건조물을 분쇄기를 이용하여 분쇄하여 분말화하여 건조엑스(15.19 g)를 얻는다. 상기 건조엑스는 -80℃ 에서 보관하여 사용한다.
Hyangbuja, Baekbokryeong, Licorice, and Huangshan are purchased on the market, and Hyangbuja, 128g, Baekbokyeong, 32g, Licorice and 32g are shredded and filtered through 2 mesh sieves. The powder is placed in an extractor, and then 1.8 L of purified water is added and extracted by heating at 95 to 100 ° C for 3 hours. The extract is filtered using a 25 μm filter and the filtrate is concentrated under reduced pressure at 50 to 60 ° C. using a concentrator. Purified water was added to the herbal residue in the same manner as above, and the extraction was repeated once to add the concentrates. The combined concentrate is put in a drier and dried at 60 ° C. or lower. The obtained dried product is pulverized using a grinder to powder to obtain a dry extract (15.19 g). The dry extract is stored at -80 ℃ to use.
[실험예][Experimental Example]
1. 실험 방법1. Experimental Method
1.1 실험동물의 준비1.1 Preparation of Laboratory Animals
실험동물은 7주령 체중 20 내지 23g의 수컷 C56BL/6 마우스를 대한 바이오링크(Daehan BioLink Inc, 한국)에서 구매하였다. 실험동물은 두마리씩 각각 짝을 이루어, 보통의 플라스틱 우리에서 식이섭취 제한 없이 생활하였다. 온도와 습도를 조절하고 12시간 간격으로 밤과 낮을 제공하였다. 또한, 대조군으로 한배새끼(Littermate)을 준비하여 일반적인 먹이를 제공하였다.
Experimental animals were purchased from Biolink (Daehan BioLink Inc, South Korea) for male C56BL / 6 mice weighing 20-23 g at 7 weeks of age. Two experimental animals were paired and lived without restriction on dietary intake in ordinary plastic cages. Temperature and humidity were controlled and provided night and day at 12 hour intervals. In addition, a litter mat (Littermate) was prepared as a control to provide a general food.
1.2 신체구금 스트레스 (1.2 Body detention stress ( RsstraintRsstraint StressStress , , RSTRST ))
5일 간의 적응 기간(Habituation)이 지난 후(도 1 참조), 신체 구금 스트레스 대조군으로 배당된 실험동물을 제외하고 통풍이 잘되는 50㎖의 폴리프로필렌 원뿔형 튜브에 실험 동물의 머리부터 넣고, 3㎝ 길이 및 15㎖의 원뿔형 튜브를 이용하여 동물의 몸을 전혀 움직일 수 없도록 고정한 후 입구를 막아 신체 구금 스트레스를 가하였다. 상기 신체 구금 상태는 오전 10시부터 하루에 2시간씩 연속 14일 동안 가해졌다(도 1의 RST 기간 참조). After a five-day habituation (see Figure 1), place the head of the experimental animal in a well ventilated 50 ml polypropylene conical tube, except for the experimental animal assigned to the physical detention stress control, starting at 3 cm And 15ml conical tube was fixed so as not to move the animal body at all after blocking the inlet was subjected to body detention stress. The detention of the body was applied for 14 consecutive days for 2 hours a day starting at 10 am (see RST period in FIG. 1).
각 신체 구금 기간이 끝나면, 스트레스를 받은 실험 동물은 각각 짝을 이루어 식이 섭취의 제한이 없는 플라스틱 우리로 돌려 보내졌다. 달리 기재되지 않은 한, 실험 동물은 마지막 행동실험 다음날 희생되었다.
At the end of each detention period, the stressed experimental animals were paired and returned to plastic cages with no dietary restrictions. Unless otherwise stated, experimental animals were sacrificed the day after the last behavioral experiment.
1.3 약물 투여1.3 Drug Administration
상기 신체 구금 기간을 마친 실험 동물들은 무작위로 무투여군(n=8), IM군(n=10), 실험군(n=12)군으로 배정하였다. 신체 구금 스트레스 기간 다음날부터 희생되는 날까지 각각 무투여군은 일반적인 식이를 제공하고, IM군은 개체당 30mg/Kg/day의 이미프라민(imipramine)을 섭취하도록 제공하고, 실험군은 개체당 400mg/Kg/day의 상기 실시예1을 섭취하도록 제공하였다(도 1의 treatment 기간 참조). Experimental animals that completed the period of physical detention were randomly assigned to the no-administration group (n = 8), IM group (n = 10), and experimental group (n = 12). From the day after the body detention stress period to the day of sacrifice, each no-administration group provided a general diet, the IM group provided 30 mg / Kg / day of imipramine per subject, and the experimental group received 400 mg / Kg per subject. Example 1 of / day was provided for ingestion (see treatment period in FIG. 1).
상기 제공한 실시예의 섭취량은 전환공식에 따르면, 사람에게서 32.5mg/Kg/day(또는 1.95g/60Kg/day)에 상당하는 것이다. 또한 신체 구금 스트레스 대조군은 일반적인 식이를 제공하였다. 전체적인 실험설계는 도 1에 도시하였다.
The intake of the above provided example corresponds to 32.5 mg / Kg / day (or 1.95 g / 60 Kg / day) in humans, according to the conversion formula. Body detention stress control also provided a general diet. The overall experimental design is shown in FIG.
1.4 행동실험 및 분석실험의 수행 및 분석1.4 Conduct and analysis of behavioral experiments and assays
본 발명에 따른 실시예의 항우울 효과를 확인하기 위하여 하기 상술하는 행동실험 및 분석실험을 수행하였다. 실험 결과의 통계분석을 위하여, 데이터들을 Graph Pad Prism 4 (GraphPad Software, USA)를 이용해 분석하였다. 또한, Newman-Keuls multiple range test와 one-way ANOVA를 이용해 다중분석을 하였다. 실험결과는 평균값 표준오차로 표시하였고, 따로 언급하지 않은 한, 통계적 오차는 5% 범주에서 인정하였다(P < 0.05).
In order to confirm the antidepressant effect of the embodiment according to the present invention, the behavioral test and analysis experiment described below were performed. For statistical analysis of the experimental results, the data were analyzed using Graph Pad Prism 4 (GraphPad Software, USA). In addition, multiple analysis was performed using Newman-Keuls multiple range test and one-way ANOVA. The experimental results were expressed as mean standard error, and unless otherwise stated, statistical errors were accepted in the 5% category (P <0.05).
2. 행동실험 결과(2. Behavior test results (
BehavioralBehavioral
test) test)
2.1. 꼬리 매달기 실험(2.1. Tail hanging experiment ( TailTail suspensionsuspension testtest ))
신체 구금 기간 종료 후 15 내지 16일째 날에 꼬리 매달기 실험을 시행하였다. 방음 처리된 방에서, 각각의 실험 마우스들의 꼬리를 50 ㎝ 높이의 금속 막대에 매달고, 테이블에는 안전매트를 깔았다. 6분간 실험이 진행되는 동안, 접착 테이프를 사용해 꼬리의 끝을 고정하였고, 6분 동안에 움직이지 않은 부동의 시간을 측정되었다. 매번 실험시, 55-60 dB의 백색소음으로 소음을 차단했다.Tail hanging experiments were conducted 15 days after the end of the physical detention period. In the soundproofed rooms, the tails of each experimental mouse were hung on a 50 cm high metal bar and a safety mat was placed on the table. During the 6 minute experiment, the tip of the tail was fixed with adhesive tape and the time of immobility that was not moved for 6 minutes was measured. In each experiment, the noise was cut off by 55-60 dB of white noise.
꼬리 매달기 실험결과는 도 2에 도시하였다. 대조군은 신체 구금 스트레스 후 정상식이, 무투여는 신체 구금 스트레스후 정상식이, 실험군은 신체 구금 스트레스후 실시예 첨가식이, IM은 신체 구금 스트레스후 이미프라민(imipramine) 첨가식이를 제공한 군이다.The tail suspension test results are shown in FIG. 2. The control group is a normal diet after body detention stress, the non-administration is a normal diet after body detention stress, the experimental group is an example diet added after body detention stress, IM is a group provided with an imipramine diet after body detention stress.
상기 실험 결과, 실험군이 무투여군보다 유의적으로 부동시간이 감소하였고, 스트레스를 받지 않은 대조군이나 항우울제를 투여한 IM군과 대비하여 유의적으로 차이가 없는 부동 시간이 나타남에 따라, 유의미한 항우울 효과가 나타남을 알 수 있다(P < 0.05 수준).
As a result of the experiment, the experimental group was significantly reduced in immobility time compared to the no-administration group, and showed a significant difference in immobility time compared to the control group that was not stressed or the IM group to which the antidepressant was administered. It can be seen that (P <0.05 level).
2.2 강제 수영 실험(2.2 Forced Swim Experiment ForcedForced swimswim testtest ))
신체 구금 스트레스 기간 종료 후 15 내지 16일째 날에 강제 수영 실험을 시행하였다. 24℃의 물이 14 cm 높이로 채워진 플렉시 유리 실린더(높이: 27cm, 지름: 15cm)에 15분간 마우스를 넣고 꺼내 말리고, 다시 우리에 넣는다. 하루 뒤에 다시 6분 동안 마우스를 실린더에 넣는다. 처음 1분 동안은 마우스들이 적응하도록 두고, 이후 5분간의 행동을 관찰하여 기록한다. 부동의 시간은 마우스들이 모든 다리를 전혀 움직이지 않고 물 위에 떠있는 총 시간이다. 매 실험시, 55-60 dB의 백색소음으로 소음을 차단했다.Forced swimming experiments were performed 15 days after the end of the physical detention stress period. Put the mouse in a Plexiglass cylinder (height: 27 cm, diameter: 15 cm) filled with 14 cm of water at 24 ° C. for 15 minutes, take it out, dry it and put it back into the cage. One day later, the mouse is placed in the cylinder for six minutes. Leave the mice to acclimate for the first 1 minute and then record and observe 5 minutes of behavior. The floating time is the total time the mice are floating on the water without moving all their legs at all. In each experiment, the noise was cut out by 55-60 dB of white noise.
강제 수영 실험결과는 도 3에 도시하였다. 대조군은 신체 구금 스트레스후 정상식이, 무투여군은 신체 구금 스트레스후 정상식이, 실험군은 신체 구금 스트레스후 실시예 첨가식이, IM군은 신체 구금 스트레스후 이미프라민(imipramine) 첨가식이를 제공한 군이다. The forced swimming test results are shown in FIG. 3. The control group is a normal diet after body detention stress, the non-administrative group is a normal diet after body detention stress, the experimental group is an example diet added after body detention stress, the IM group is a group provided with an imipramine diet after body detention stress. .
상기 실험 결과, 실험군이 무투여군보다 유의적으로 부동시간이 감소하였고, 스트레스를 받지 않은 대조군이나 항우울제를 투여한 IM군과 대비하여 유의적으로 차이가 없는 부동 시간이 나타남에 따라, 유의미한 항우울 효과가 나타남을 알 수 있다(P < 0.05 수준).
As a result of the experiment, the experimental group was significantly reduced in immobility time compared to the no-administration group, and showed a significant difference in immobility time compared to the control group that was not stressed or the IM group to which the antidepressant was administered. It can be seen that (P <0.05 level).
2.3 새로운 2.3 new 환경내Within the environment 섭식feeding 억제 실험 ( Inhibition experiment ( NoveltyNovelty suppressedsuppressed feedingfeeding ))
신체 구금 기간 종료 후 17 내지 18일째 날에 새로운 환경내 섭식 억제(novelty suppressed feeding) 실험을 시행하였다. 이 실험은 동물이 먹이에 접근하기까지 걸리는 시간을 측정하여, 스트레스로 유발된 불안의 정도를 평가하는 것으로, 만성적인 항우울 처치에 민감하다. A new novelty suppressed feeding experiment was conducted 17 to 18 days after the end of the physical detention period. This experiment measures the time it takes for an animal to reach its prey and evaluates the degree of stress-induced anxiety and is sensitive to chronic antidepressant treatment.
실험 전, 동물에게 물은 제한없이 제공되지만, 먹이는 24시간 동안 제거한다. 이 시기의 마지막에, 개방된 공간(45×45×40cm)의 중앙에 원형의 흰 거름종이(지름 150mm)를 놓고, 그 위에 하나의 2×2.5cm 타원형의 고체 사료를 놓는다. 각각의 마우스를 원래 있던 우리에서 빼내어, 사료와 물이 없는 우리에서 30분간 굶긴 후에, 조도 1400 lux의 개방된 공간의 구석에 놓아준다. 5분 동안 고체 사료를 처음으로 깨물 때까지 걸리는 잠재 기간을 측정한다. 마우스가 먹이를 먹기 시작한 즉시, 그들을 원래 있었던 우리로 옮겨서 계량된 사료조각과 함께 5분 동안 놓아둔다. 이 기간의 마지막에 남은 고체 사료의 무게를 측정하여, 먹이소비량을 계산한다. 매 실험시, 55-60 dB의 백색소음으로 소음을 차단했다.Before the experiment, animals are given unlimited water, but food is removed for 24 hours. At the end of this period, a circular white filter paper (150 mm in diameter) is placed in the center of the open space (45 x 45 x 40 cm) and on top of it is a single 2 x 2.5 cm oval solid feed. Each mouse is removed from its original cage, starved for 30 minutes in a cage without food and water, and then placed in the corner of an open space of 1400 lux illuminance. The potential period of time until the first bite of the solid feed for 5 minutes is measured. As soon as the mice begin to feed, they are transferred to their original cages and left for 5 minutes with weighed feed pieces. At the end of this period, the remaining solid feed is weighed to calculate the food consumption. In each experiment, the noise was cut out by 55-60 dB of white noise.
상기 새로운 환경내 섭식 억제 실험결과는 도 4에 도시하였다. 대조군은 신체 구금 스트레스후 정상식이, 무투여군은 신체 구금 스트레스후 정상식이, 실험군은 신체 구금 스트레스후 실시예 첨가식이, IM군은 신체 구금 스트레스후 이미프라민(imipramine) 첨가식이를 제공한 군이다. The results of the new environment intake inhibition experiment are shown in FIG. 4. The control group is a normal diet after body detention stress, the non-administrative group is a normal diet after body detention stress, the experimental group is an example diet added after body detention stress, the IM group is a group provided with an imipramine diet after body detention stress. .
상기 실험 결과, 실험군이 무투여군보다 유의적으로 식이 시간이 감소하였고, 신체 구금 스트레스를 받지 않은 대조군이나 항우울제를 투여한 IM군과 대비하여 유의적인 차이가 나타나지 않아, 특히 만성적인 우울증에 대한 항우울 효과가 우수한 것을 알 수 있다(P < 0.05 수준).
As a result of the experiment, the experimental group significantly reduced the diet time than the no-administrative group, and did not show a significant difference compared to the control group that did not receive physical detention stress or the IM group that received antidepressant, especially antidepressant for chronic depression It can be seen that the effect is excellent (P <0.05 level).
2.4 구슬 묻기 실험 (2.4 Bead Buried Experiment MarbleMarble buryingburying testtest ))
신체 구금 스트레스 기간 종료 후 17 내지 18 일째 날에 구슬 묻기 실험을 시행하였다. 마우스의 파묻기(burying)는 방어 행동으로서 작용하고, 위협의 강도 또는 위협에 대한 동물의 취약성을 감소시키는데 사용되어, 구슬 묻기 실험은 불안을 측정하기 위한 모델로써 사용된다. 빈 우리에 푹신한 것들을 5cm 정도 깔고, 12개의 구슬을 우리 전체에 평평하게 놓아둔다. 30분 동안 마우스들이 우리를 자유롭게 돌아다니게 한 후에, 성공적으로 묻혀진 구슬의 개수를 센다. 25% 이하의 구슬이 표면에서 보일 때를 묻혔다고 정의한다. 매 실험시, 55-60 dB의 백색소음으로 소음을 차단했다.Bead bubbling experiments were conducted on days 17-18 after the end of the physical detention stress period. Burying of the mouse acts as a defensive action and is used to reduce the intensity of the threat or the animal's vulnerability to the threat, so the marble buried experiment is used as a model for measuring anxiety. Lay the fluffy things in the empty cage about 5cm and place the 12 beads flat throughout us. After 30 minutes let the mice roam freely, count the number of successfully buried marbles. It is defined that less than 25% of the beads are buried when seen from the surface. In each experiment, the noise was cut out by 55-60 dB of white noise.
상기 구슬 묻기 실험결과는 도 5에 도시하였다. 대조군은 신체 구금 스트레스후 정상식이, 무투여군은 신체 구금 스트레스후 정상식이, 실험군은 신체 구금 스트레스후 실시예 첨가식이, IM군은 신체 구금 스트레스후 이미프라민(imipramine) 첨가식이를 제공한 군이다. The results of the buried beads are shown in FIG. 5. The control group is a normal diet after body detention stress, the non-administrative group is a normal diet after body detention stress, the experimental group is an example diet added after body detention stress, the IM group is a group provided with an imipramine diet after body detention stress. .
상기 실험 결과, 실험군이 무투여군보다 구슬을 묻은 수가 유의적으로 증가하였고, 신체 구금 스트레스를 받지 않은 대조군이나 항우울제를 투여한 IM군과 대비하여 유의적인 차이가 나타나지 않아, 특히 만성적인 우울증에 대한 항우울 효과가 유의적으로 우수한 것을 알 수 있다(P < 0.05 수준).
As a result of the experiment, the number of beads buried in the experimental group was significantly increased than the non-administrative group, and there was no significant difference compared to the control group which did not receive physical detention stress or the IM group which received antidepressant. It can be seen that the depression effect is significantly superior (P <0.05 level).
3. 세포배양 실험3. Cell Culture Experiment
3.1 세포 생존율 측정 (3.1 Measuring Cell Viability (
신경독성평가Neurotoxicity Assessment
))
1) 세포 배양1) Cell culture
SH-SY5Y 신경아세포를 Dulbeccos modified Eagles medium(DMEM; Gibco-BRL, USA)에 10% 열불활성화된 우태아혈청(heat-inactivated fetal bovine serum, Gibco-BRL. USA)과 페니실린(penicillin) 20 units/ml, 및 스트렙토마이신(streptomycin) 20 mg/ml을 섞은 배지에서 37℃, 95% 공기와 5% CO2 환경의 가습배양기에서 배양하였다. SH-SY5Y neuroblasts were treated with 10% heat-inactivated fetal bovine serum (Gibco-BRL.USA) and penicillin in Dulbeccos modified Eagles medium (DMEM; Gibco-BRL, USA). ml and 20 mg / ml of streptomycin were incubated in a humidified incubator at 37 ° C., 95% air and 5% CO 2 .
과산화수소(H2O2, Sigma Aldrich, USA)를 순차적으로 물에 용해시켜 사용하였으며, 세포들은 무혈청 DMEM에 H2O2와 함께 실험군 배당에 따라 실시예1을 첨가(50㎎/㎖, 100㎎/㎖)한 배지에서 배양하였다. 대비되는 그룹으로 H2O2 비처리군, H2O2 처리후 실시예 무첨가군, 비타민 C 첨가군(200mM), 트롤록스(TRX, Trolox, 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid)) 첨가군(200mM)를 사용하였다.
Hydrogen peroxide (H 2 O 2 , Sigma Aldrich, USA) was used to sequentially dissolve in water, cells were added to Example 1 according to the experimental group dividend with H 2 O 2 in serum-free DMEM (50 mg / ㎖, 100 Mg / ml) in a medium. In contrast to the H 2 O 2 non-treated group, H 2 O 2 treatment-free group, vitamin C added group (200mM), trolox (TRX, Trolox, 6-hydroxy-2,5,7,8- tetramethylchroman-2-carboxylic acid)) addition group (200 mM) was used.
2) 세포 생존율 측정 2) Cell viability measurement
신경독성은 WST-1 assay를 통해 세포 생존력을 평가하여 측정하였으며, Cell proliferation reagent WST-1(Roche) 사용하였다. 상기 실험군 배당에 따라 실시예1, 비타민 C 또는 트롤록스를 배지에 첨가하고, 800mM의 과산화수소를 가한 후 24시간 후에 생존 세포의 수를 분석하였다. 테트라졸리움염(Tetrazolium salt) WST-1 용액을 각 well내 배지의 1/10 부피(v/v)로 첨가한 후, 5%의 CO2 및 37℃의 가습 배양기에서 2시간 동안 배양하여 측정하였다.Neurotoxicity was measured by assessing cell viability through the WST-1 assay, and cell proliferation reagent WST-1 (Roche) was used. According to the experimental group divide, Example 1, vitamin C or trolox was added to the medium, and the number of viable cells was analyzed 24 hours after adding 800 mM hydrogen peroxide. Tetrazolium salt WST-1 solution was measured by adding 1/10 volume (v / v) of the medium in each well, followed by incubation for 2 hours in a humidified incubator at 37% with 5% CO 2. .
테트라졸리움염(Tetrazolium salt) WST-1은 미트콘드리아 효소인 숙신산탈수소효소(succinate-dehμydrogenase)에 의해서 수용성 검붉은 포르마잔(formazan)으로 변하고, 이는 생존 세포의 수에 비례한다. 이후 배지를 제거하고, 생존력을 측정하기 440nm에서 여기(excitation)의 흡광도를 측정하고, 560nm에서 방출기(emission)의 흡광도를 측정하였다.Tetrazolium salt WST-1 is converted to water-soluble dark red formazan by the mitochondrial enzyme succinate-dehμydrogenase, which is proportional to the number of viable cells. The medium was then removed, and the absorbance of excitation was measured at 440 nm to measure viability, and the absorbance of the emission at 560 nm.
상기 세포 생존 실험 결과는 도 6에 도시하였다. (-,0)은 과산화수소도 투여하지 않은 군이고, (+,0)은 과산화수소만 투여한 군, (+.50)은 과산화수소 및 50㎎/㎖ 실시예1 투여한 군, (+,100)은 과산화수소 및 100㎎/㎖ 실시예1 투여한 군, (+,VitC)는 과산화수소 및 비타민 C 투여한 군, (+,TRX)는 과산화수소 및 트롤록스를 투여한 군이다. The cell survival test results are shown in FIG. 6. (-, 0) is the group not administered hydrogen peroxide, (+, 0) is the group administered only hydrogen peroxide, (+.50) is the group administered with hydrogen peroxide and 50 mg / ml Example 1, (+, 100) Silver hydrogen peroxide and 100 mg / ml Example 1 group, (+, VitC) is a group administered hydrogen peroxide and vitamin C, (+, TRX) is a group administered hydrogen peroxide and trolox.
상기 실험 결과, 과산화수소에 의한 세포 수의 감소를 본 발명에 의한 실시예가 유의적으로 억제하고, 세포 보호 효과는 실시예의 농도가 높을 수록 증가함을 알 수 있다. 특히 본 발명에 따른 실시예를 100㎎/㎖ 투여한 군은 실시예를 투여하지 않은 군보다 유의적으로 세포를 보호하였으며, 비타민 C나 트롤록스를 투여한 군과 대비하여 유의적으로 차이가 없는 정도의 세포 보호 효과를 갖는 것으로 나타났다(P < 0.01 수준).
As a result of the experiment, it can be seen that the embodiment according to the present invention significantly inhibits the decrease in the number of cells due to hydrogen peroxide, and the cell protective effect increases as the concentration of the example increases. In particular, the group administered 100 mg / ㎖ according to the present invention significantly protected the cells than the group not administered the example, there is no significant difference compared to the group administered with vitamin C or trolox It was shown to have a degree of cytoprotective effect (P <0.01 level).
3.2 3.2 항산화력Antioxidant power 실험 Experiment
SH-SY5Y 세포를 96-well plate에 담아, 상기 실험군 배당에 따라 실시예1, 비타민 C 또는 트롤록스를 배지에 첨가하고 200μM의 과산화수소를 가한 후 24시간 동안 배양하여 세포내 활성 산소종(reactive oxygen species, ROS)에 대한 인지자를 사용하여 형광분석하였다. 형광분석을 위해 CM-H2DCFDA (5-(and-6)-chloromethyl-2',7'-dichlorodihydro fluorescein diacetate. Invitrogrn), 과산화수소 관련 활성 산소종(hydrogen peroxide-related ROS)에 민감한 세포막 투과성의 형광 염료(DCF; Molecular Probes, USA), 세포막 투과성 과산화물에 민감한 형광염료(molecular probes)들을 이용하여 활성 산소종 발현을 평가하여 항산화력을 측정하였다. In a 96-well plate, SH-SY5Y cells were added to the medium according to Example 1, vitamin C or trolox, and 200 μM hydrogen peroxide was added to the medium, followed by incubation for 24 hours. fluorescence using a recognizer for species, ROS). Cell membrane permeability sensitive to CM-H 2 DCFDA (5- (and-6) -chloromethyl-2 ', 7'-dichlorodihydro fluorescein diacetate.Invitrogrn) and hydrogen peroxide-related ROS for fluorescence analysis Antioxidant activity was measured by evaluating reactive oxygen species expression using fluorescent dyes (DCF; Molecular Probes, USA) and fluorescent dyes (molecular probes) sensitive to cell membrane permeable peroxides.
보다 구체적으로, 상기 배양한 세포를 10μM CM-H2DCFDA 처리하고 37℃에서 30분 동안 배양하고, 헤페스 완충액(Hepes buffer)으로 3회 씻어 내어 형광분석하였다. 형광 분광 광도계(SpectraMax M5; Molecular Devices Corporation, USA)를 이용해서, 488nm에서 여기(excitation)의 형광수준을 측정하고, 510nm에서 방출기(emission)의 형광 수준을 측정하였고, 3번 측정의 평균값을 계산하여 단일 측정값으로 간주하였다.More specifically, the cultured cells were treated with 10 μM CM-H 2 DCFDA, incubated at 37 ° C. for 30 minutes, washed three times with Hepes buffer, and fluorescence. Using a fluorescence spectrophotometer (SpectraMax M5; Molecular Devices Corporation, USA), the fluorescence level of excitation was measured at 488 nm, the fluorescence level of the emission was measured at 510 nm, and the average value of the three measurements was calculated. As a single measurement.
상기 항산화력 실험 결과는 도 7에 도시하였다. (-,0)은 과산화수소도 투여하지 않은 군이고, (+,0)은 과산화수소만 투여한 군, (+.50)은 과산화수소 및 50㎎/㎖ 실시예1 투여한 군, (+,100)은 과산화수소 및 100㎎/㎖ 실시예1 투여한 군, (+,VitC)는 과산화수소 및 비타민 C 투여한 군, (+,TRX)는 과산화수소 및 트롤록스를 투여한 군이다. The antioxidant test results are shown in FIG. 7. (-, 0) is the group not administered hydrogen peroxide, (+, 0) is the group administered only hydrogen peroxide, (+.50) is the group administered with hydrogen peroxide and 50 mg / ml Example 1, (+, 100) Silver hydrogen peroxide and 100 mg / ml Example 1 group, (+, VitC) is a group administered hydrogen peroxide and vitamin C, (+, TRX) is a group administered hydrogen peroxide and trolox.
상기 실험 결과, 과산화수소에 의한 활성 산소종 생성을 본 발명에 의한 실시예가 유의적으로 억제하고, 항산화 효과는 실시예의 농도가 높을 수록 증가함을 알 수 있다. 특히 본 발명에 따른 실시예를 100㎎/㎖ 투여한 군은 실시예를 투여하지 않은 군보다 유의적으로 세포를 보호하였으며, 비타민 C나 트롤록스를 투여한 군 보다도 더욱 억제하는 효과를 갖는 것으로 나타났다(P < 0.01 수준).
As a result of the experiment, it can be seen that the embodiment according to the present invention significantly inhibits the generation of active oxygen species by hydrogen peroxide, and the antioxidant effect increases as the concentration of the example increases. In particular, the group administered 100 mg / ㎖ according to the present invention significantly protected the cells than the group not administered the example, it was shown to have a more inhibitory effect than the group administered with vitamin C or trolox (P <0.01 level).
4. 산화 스트레스 측정4. Oxidative Stress Measurement
신체 구금 스트레스에 의한 뇌의 산화 스트레스를 지질과산화물인 말론다이알데하이드(Malondialdehyde, MDA) 측정을 통해 평가하였다. 실험 마우스들을 희생한 후 뇌의 해마를 분리하여, 5 mM 부틸레이트하이드록시톨루엔(butylated hydroxytoluene, BHT)을 함유하는 차가운 20 mM 인산완충식염수(phosphate-buffered saline, PBS, pH 7.4)를 조직부피의 4배 부피로 가하여 균질화 하였다. 상기 균질액을 3,000 g에서 4℃, 10분간 원심 분리하여 상청액을 수득하여 분석하였다. Oxidative stress in the brain caused by stress in body detention was assessed by measuring malondialdehyde (MDA), a lipid peroxide. After sacrificing experimental mice, the hippocampus of the brain was isolated, and cold 20 mM phosphate-buffered saline (PBS, pH 7.4) containing 5 mM butylated hydroxytoluene (BHT) was added to the tissue volume. Homogenized by adding 4 times volume. The homogenate was centrifuged at 3,000 g at 4 ° C. for 10 minutes to obtain a supernatant and analyzed.
상기 상청액 200㎕를 probucol 10㎕, Bioxytech MDA-586 kit(Oxis Research, USA)에서 제공된 희석된 R1 reagent(methanol:N-methyl-2-phenylindole = 1:3) 640㎕ 및 12N HCl 150㎕와 섞는다. 각각을 45℃에서 60분 동안 배양한 후, 10,000g으로 10분 동안 원심분리 하였다. 586 nm에서 상청액의 흡광도를 측정하였다. MDA 측정값을 단백질 농도를 기준으로 표준화하고, 샴대조값(sham control value)의 백분율로 표현하였다. 단백질과 결합된 MDA의 양은 뺄셈에 의하여 계산되었다. 단백질 농도는 브래드퍼드법(Bradford method)에 의해 결정하였다(Bio-Rad Laboratories, USA).200 μl of the supernatant is mixed with 10 μl of probucol, 640 μl of diluted R1 reagent (methanol: N-methyl-2-phenylindole = 1: 3) and 150 μl of 12N HCl provided by Bioxytech MDA-586 kit (Oxis Research, USA). . Each was incubated at 45 ° C. for 60 minutes and then centrifuged at 10,000 g for 10 minutes. The absorbance of the supernatant was measured at 586 nm. MDA measurements were normalized based on protein concentration and expressed as a percentage of sham control value. The amount of MDA bound to the protein was calculated by subtraction. Protein concentration was determined by the Bradford method (Bio-Rad Laboratories, USA).
상기 항산화력 실험 결과는 도 8에 도시하였다. 대조군은 신체 구금하지 않고 정상식이를 제공한 군이고, 무투여+구속군은 신체 구금한 후 정상식이를 제공한 군이고, 실험군+구속군은 신체 구금한 후 실시예를 첨가한 식이를 제공한 군이다. The antioxidant test results are shown in FIG. 8. The control group was a group that provided a normal diet without body detention, and the group without administration + restraint was a group who provided a normal diet after detaining the body, and the experimental group + group was provided with a diet added to the Example after detaining the body. It's a military.
상기 실험 결과, 신체 구금 스트레스를 가한 경우 해마에서의 말론디알데하이드 생성이 유의적으로 증가하는데(P < 0.01), 본 발명에 의한 실시예를 첨가한 식이를 섭취한 군에서 말론디알데하이드 생성이 유의적으로 감소한 것을 알 수 있다(P < 0.05 수준).
As a result of the above experiment, malondialdehyde production was significantly increased in the hippocampus when body detention stress was applied (P <0.01), but malondialdehyde production was significantly increased in the group ingesting the diet to which the embodiment according to the present invention was added. It can be seen that the decrease was (P <0.05 level).
상기 상술한 세포배양 및 항산화력 실험결과로부터, 본 발명에 따른 실시예는 세포 보호 효과 및 항산화효과를 통해 우울증을 예방하고 치료하는 효과가 있음을 확인할 수 있다.
From the above-described cell culture and antioxidant power test results, it can be confirmed that the embodiment according to the present invention has an effect of preventing and treating depression through a cell protective effect and an antioxidant effect.
대한약전 제제총칙 중 산제의 제조방법에 따라 1포당 하기의 성분 함량으로 혼합하고 기밀포에 충진하여 제조한다.According to the preparation method of powder in Korean Pharmacopoeia General Formulation, it is prepared by mixing with the following ingredient content per bag and filling into airtight cloth.
건조분말(실시예1) - 200㎎Dry Powder (Example 1)-200 mg
유당 - 100㎎Lactose-100 mg
탈크 - 10㎎
Talc-10 mg
대한약전 제제총칙 중 정제의 제조방법에 따라 1 정당 하기의 성분 함량으로 혼합한 후, 통상의 정제의 제조방법에 따라 타정하여 정제를 제조한다.In accordance with the manufacturing method of the tablet of the Korean Pharmacopoeia General Formulation, the mixture is mixed in the amounts of 1 to 1, and then tableted according to the manufacturing method of a conventional tablet to prepare a tablet.
건조분말(실시예1) - 100㎎Dry Powder (Example 1)-100 mg
옥수수전분 - 100㎎Corn Starch-100mg
유당 - 100㎎Lactose-100 mg
스테아린산 마그네슘 - 2㎎
Magnesium Stearate-2mg
대한약전 제제총칙 중 캅셀제의 제조방법에 따라 1캅셀당 하기의 성분 함량으로 혼합하고 젤라틴 캡슐에 충전하여 캡슐제를 제조한다.According to the preparation method of capsules in the Korean Pharmacopoeia General Formulation, the capsules are prepared by mixing the amount of the following ingredients per capsule and filling the gelatin capsules.
건조분말(실시예1) - 100㎎Dry Powder (Example 1)-100 mg
결정성 셀룰로오스 - 30㎎Crystalline Cellulose-30mg
유당 - 100㎎Lactose-100 mg
스테아린산 마그네슘 - 2㎎
Magnesium Stearate-2mg
대한약전 제제총칙 중 액제의 제조방법에 따라 액제 하기의 성분 함량으로 가하여 용해시키고 레몬향을 적량 가한 다음 정제수를 가하여 전체 100㎖가 되도록 조절한 후 갈색병에 충진하여 멸균하여 제조한다.According to the preparation method of the liquid formulation of the Korean Pharmacopoeia General Formulation, the solution is added to the following ingredients to dissolve, add lemon flavor, and then add purified water to adjust the total volume to 100 ml.
건조분말(실시예1) - 200㎎Dry Powder (Example 1)-200 mg
이성화당 - 100㎎Isomerized Sugar-100mg
만니톨 - 10㎎Mannitol-10mg
정제수 - 적량
Purified Water-Appropriate
통상의 기능성 식품(건강기능식품) 제조방법에 따라 하기의 성분을 혼합하여 건강기능식품을 제조한다. 하기의 비타민 및 미네랄 혼합물의 조성비는 비교적 건강기능식품에 적합한 성분을 영양권장량에 준하여 제형예로 조성하였으나, 그 배합비를 임의로 변형하여 실시하여도 무방하다. According to the conventional functional food (health functional food) manufacturing method by mixing the following ingredients to produce a health functional food. The composition ratio of the following vitamin and mineral mixtures was formulated as an example of the formulation based on the nutritional recommended amount of a relatively suitable component for health functional food, but may be carried out by modifying the formulation ratio arbitrarily.
건조분말(실시예1) - 1000㎎Dry Powder (Example 1)-1000 mg
비타민 혼합물 - 적량 Vitamin Mixture-Appropriate
(비타민 A 아세테이트 70㎍. 비타민 E 1㎎, 비타민 B1 0.2㎎, 비타민 B2 0.2㎎, 비타민 B6 0.2㎎, 나이아신 20㎎NE, 엽산 50㎍)(70 μg of Vitamin A
무기질 혼합물 - 적량Mineral Mixture-Correct
(황산제1철 2㎎, 산화아연 1㎎, 탄산마그네슘 20㎎,인산칼슘 100㎎, 구연산칼륨 100㎎, 염화마그네슘 25㎎)
(
통상의 음료 제조방법에 따라 하기의 성분을 혼합한 다음, 약 1시간 내지 2시간 동안 가열교반한 다음 멸균한 다음 멸균된 음료 용기에 주입밀봉하여 기능성음료(액상 건강기능식품)을 제조하였다. 하기의 조성비는 비교적 음료 조성에 적합한 성분비율로 조성하였으나, 그 배합비를 임의로 변형하여 실시하여도 무방하다. According to the conventional beverage preparation method, the following ingredients were mixed, heated and stirred for about 1 hour to 2 hours, and then sterilized and injected and sealed in a sterile beverage container to prepare a functional beverage (liquid health functional food). Although the following composition ratio was comprised by the component ratio which is comparatively suitable for a beverage composition, you may carry out by modifying the compounding ratio arbitrarily.
건조분말(실시예1) - 1000㎎Dry Powder (Example 1)-1000 mg
구연산 - 1000㎎Citric Acid-1000 mg
올리고당 - 100gOligosaccharide-100 g
타우린 - 1gTaurine-1 g
정제수 - 적량
Purified Water-Appropriate
이상으로 본 발명 내용의 특정한 부분을 상세히 기술하였는 바, 당업계의 통상의 지식을 가진 자에게 있어서, 이러한 구체적 기술은 단지 바람직한 실시 양태일 뿐이며, 이에 의해 본 발명의 범위가 제한되는 것이 아닌 점은 명백할 것이다. 따라서 본 발명의 실질적인 범위는 첨부된 청구항 들과 그것들의 등가물에 의하여 정의된다고 할 것이다.While the present invention has been particularly shown and described with reference to specific embodiments thereof, those skilled in the art will appreciate that such specific embodiments are merely preferred embodiments and that the scope of the present invention is not limited thereby. something to do. It is therefore intended that the scope of the invention be defined by the claims appended hereto and their equivalents.
Claims (18)
상기 향부자, 백복령, 감초 및 황련의 추출물은 향부자 : 백복령 : 감초 : 황련의 중량비율이 2 내지 6 : 1 내지 2 : 1 내지 2 : 1 내지 2 인 것을 특징으로 하는 우울증 예방 및 치료용 약학조성물.The method of claim 1,
The extract of the hyangbuja, baekbokyeong, licorice and sulfur lysine is the weight ratio of hyangbuja: baekbokyeong: licorice: huangyeon 2 to 6: 1 to 2: 1 to 2: characterized in that the pharmaceutical composition for preventing and treating depression.
상기 향부자, 백복령, 감초 및 황련의 추출물은 향부자 : 백복령 : 감초 : 황련의 중량비율이 4 : 1 : 1 : 1 인 것을 특징으로 하는 우울증 예방 및 치료용 약학조성물.The method of claim 1,
The extract of the hyangbuja, baekbokyeong, licorice and Huangren is a hyangbuja: baekbokyeong: licorice: the weight ratio of the rhubarb 4: 1: 1: 1 pharmaceutical composition for the prevention and treatment of depression.
상기 추출물은 정제수를 포함한 물, 탄소수 1 내지 탄소수 4의 저급 알코올 또는 이들의 혼합용매로부터 선택된 용매로 가용한 추출물인 것을 특징으로 하는 우울증 예방 및 치료용 약학조성물.The method of claim 1,
The extract is a pharmaceutical composition for preventing and treating depression, characterized in that the extract available in a solvent selected from water, including purified water, a lower alcohol having 1 to 4 carbon atoms or a mixed solvent thereof.
상기 약학조성물은 과립제, 레모네이드제, 산제, 시럽제, 액제, 엑스제, 엘릭서제, 유동엑스제, 유제, 현탁제, 전제, 침제, 정제, 주정제, 캡슐제, 틴크제 및 환제로 이루어진 군에서 선택된 어느 하나인 제형인 것을 특징으로 하는 우울증 예방 및 치료용 약학조성물.The method according to any one of claims 1 to 4,
The pharmaceutical composition is in the group consisting of granules, lemonades, powders, syrups, solutions, extracts, elixirs, liquid extracts, emulsions, suspensions, premises, acupuncture, tablets, spirits, capsules, tinctures and pills Pharmaceutical composition for preventing and treating depression, characterized in that the formulation is any one selected.
상기 추출물을 농축하는 단계를 포함하는 우울증 예방 및 치료용 약학조성물 제조방법.Adding a solvent to Hyangbuja, Baekbokyeong, licorice and rhubarb and extracting to obtain an extract; And
Method for producing a pharmaceutical composition for preventing and treating depression comprising the step of concentrating the extract.
상기 추출물을 수득하는 단계는 향부자, 백복령, 감초 및 황련의 총 중량대비 5 내지 20배 중량의 용매를 가하여 추출하는 것을 특징으로 하는 우울증 예방 및 치료용 약학조성물 제조방법.The method according to claim 6,
The step of obtaining the extract is a method for producing a pharmaceutical composition for preventing and treating depression, characterized in that the extract by adding a solvent of 5 to 20 times the weight relative to the total weight of Hyangbuja, Baekbokyeong, licorice and sulfur.
상기 추출물을 수득하는 단계는 80℃ 내지 120℃에서 1시간 내지 10시간 동안 추출하는 것을 특징으로 하는 우울증 예방 및 치료용 약학조성물 제조방법.The method according to claim 6,
Obtaining the extract is a pharmaceutical composition for preventing and treating depression, characterized in that the extraction for 1 hour to 10 hours at 80 ℃ to 120 ℃.
상기 농축하는 단계는 상기 추출물을 여과하는 단계; 상기 여과된 추출물을 감압 농축하는 단계, 상기 농축된 추출물을 건조하는 단계; 상기 건조된 농축물을 분쇄하여 분말화 하는 단계를 포함하는 것을 특징으로 하는 우울증 예방 및 치료용 약학조성물 제조방법.The method according to claim 6,
The step of concentrating includes filtering the extract; Concentrating the filtered extract under reduced pressure, and drying the concentrated extract; Method for preparing a pharmaceutical composition for preventing and treating depression, characterized in that it comprises the step of pulverizing the dried concentrate.
상기 향부자, 백복령, 감초 및 황련의 추출물은 향부자 : 백복령 : 감초 : 황련의 중량비율이 2 내지 6 : 1 내지 2 : 1 내지 2 : 1 내지 2 인 것을 특징으로 하는 우울증 예방 및 치료용 건강기능식품.The method of claim 10,
The extract of the hyangbuja, baekbokyeong, licorice and the yellow lotus is the weight ratio of hyangbuja: baekbokyeong: licorice: rhubarb weight ratio of 2 to 6: 1 to 2: 1 to 2: 1 to 2, characterized in that the health functional food for preventing and treating depression .
상기 향부자, 백복령, 감초 및 황련의 추출물은 향부자 : 백복령 : 감초 : 황련의 중량비율이 4 : 1 : 1 : 1 인 것을 특징으로 하는 우울증 예방 및 치료용 건강기능식품.The method of claim 10,
The extract of the hyangbuja, baekbokyeong, licorice and Huangren is a hyangbuja: baekbokyeong: licorice: the weight ratio of rhubarb is 4: 1: 1: 1 functional food for preventing and treating depression.
상기 추출물은 정제수를 포함한 물, 탄소수 1 내지 4의 저급 알코올 또는 이들의 혼합용매로부터 선택된 용매로 가용한 추출물인 것을 특징으로 하는 우울증 예방 및 치료용 건강기능식품.The method of claim 10,
The extract is a functional food for preventing and treating depression, characterized in that the extract available in a solvent selected from water, including purified water, lower alcohols having 1 to 4 carbon atoms or a mixed solvent thereof.
상기 건강기능식품은 캡슐, 정제, 분말, 과립, 액상, 환, 편상, 페이스트상, 시럽, 겔, 젤리 및 바로 이루어진 군에서 선택된 어느 하나의 제형인 것을 특징으로 하는 우울증 예방 및 치료용 건강기능식품.The method according to any one of claims 10 to 13,
The health functional food is a capsule, tablets, powders, granules, liquid, pill, flaky, paste, syrup, gel, jelly and just one of the formulations selected from the group consisting of health functional foods for preventing and treating depression .
상기 추출물을 농축하는 단계를 포함하는 우울증 예방 및 치료용 건강기능식품 제조방법.Adding a solvent to Hyangbuja, Baekbokyeong, licorice and rhubarb and extracting to obtain an extract; And
Health functional food manufacturing method for preventing and treating depression comprising the step of concentrating the extract.
상기 추출물을 수득하는 단계는 향부자, 백복령, 감초 및 황련의 총 중량대비 5 내지 20배 중량의 용매를 가하여 추출하는 것을 특징으로 하는 우울증 예방 및 치료용 건강기능식품 제조방법.16. The method of claim 15,
Obtaining the extract is a method for producing a health functional food for preventing and treating depression, characterized in that the extract by adding a solvent of 5 to 20 times the weight relative to the total weight of Hyangbuja, Paekbokyeong, licorice and sulfur.
상기 추출물을 수득하는 단계는 80℃ 내지 120℃에서 1시간 내지 10시간 동안 추출하는 것을 특징으로 하는 우울증 예방 및 치료용 건강기능식품 제조방법.16. The method of claim 15,
Obtaining the extract is a method for producing health functional food for preventing and treating depression, characterized in that the extract for 1 hour to 10 hours at 80 ℃ to 120 ℃.
상기 농축하는 단계는 상기 추출물을 여과하는 단계; 상기 여과된 추출물을 감압 농축하는 단계, 상기 농축된 추출물을 건조하는 단계; 상기 건조된 농축물을 분쇄하여 분말화 하는 단계를 포함하는 것을 특징으로 하는 우울증 예방 및 치료용 건강기능식품 제조방법.
16. The method of claim 15,
The step of concentrating includes filtering the extract; Concentrating the filtered extract under reduced pressure, and drying the concentrated extract; Method for producing a health functional food for preventing and treating depression, characterized in that it comprises the step of pulverizing the dried concentrate.
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