KR20140114950A - A composition comprising Amomum cardamomum L. extracts having anti-obesity activity - Google Patents
A composition comprising Amomum cardamomum L. extracts having anti-obesity activity Download PDFInfo
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- KR20140114950A KR20140114950A KR1020130028859A KR20130028859A KR20140114950A KR 20140114950 A KR20140114950 A KR 20140114950A KR 1020130028859 A KR1020130028859 A KR 1020130028859A KR 20130028859 A KR20130028859 A KR 20130028859A KR 20140114950 A KR20140114950 A KR 20140114950A
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- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
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- A—HUMAN NECESSITIES
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- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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Abstract
Description
본 발명은 백두구 추출물을 포함하는 항비만 조성물에 관한 것으로서, 보다 상세하게는 백두구 추출물을 유효성분으로 포함하는 비만 예방 또는 치료용 약학 조성물 및 백두구 추출물을 유효성분으로 포함하는 비만 예방 또는 개선용 건강기능 식품에 관한 것이다.
The present invention relates to an anti-obesity composition comprising an extract of Baekduguchi. More particularly, the present invention relates to a pharmaceutical composition for preventing or treating obesity comprising an Baekduguk extract as an active ingredient and a health function for preventing or improving obesity It is about food.
비만(obesity) 또는 비만증은 사람의 몸무게가 지나치게 많이 나가는 것을 통틀어 부르는 말로서 병리학적으로는 세계보건기구(WHO)에서 정한 이른바 표준 몸무게 기준치(body mass index; BMI)가 30 이상인 경우를 일컫지만 한국에서는 25 이상이면 비만으로 진단한다. 대부분의 경우 체중이 정상치보다 많이 나가는 것을 뜻하지만 체중이 그다지 많이 나가지 않더라도 몸의 구성성분 중 지방의 비율이 높은 경우 비만이라고 한다. 비만은 그 자체로서는 문제가 되지 않을 수도 있으나, 이로 인해 야기되는 사회적 장애 및 과다한 지방으로 유발되는 2차 합병증이 심각하다. 예를 들어 비만은 고지혈증, 고혈압, 동맥경화, 당뇨병, 지방간, 관절이상 등의 발생비율을 현저하게 증가시킬 수 있다.Obesity or obesity refers to the excess weight of a person's body weight, which is pathologically referred to as the World Health Organization's (WHO) standard body mass index (BMI) of 30 or more. If it is 25 or more, it is diagnosed as obesity. In most cases, weight means more than normal, but even if you do not have a lot of weight, the proportion of fat in your body is high. Obesity may not be a problem in itself, but it has serious social problems and secondary complications caused by excessive fat. For example, obesity can significantly increase the incidence of hyperlipidemia, hypertension, arteriosclerosis, diabetes, fatty liver, joint disorders, and the like.
이와 같이 비만은 각종 성인병의 원인이 된다. 체중이 늘면 혈당처리 능력이 떨어져 당뇨병을 일으키기 쉬우며 고혈압도 발생할 수 있다. 체중이 정상체중보다 20% 증가되면 당뇨병 발병 가능성은 정상인의 10배로 증가한다. 또한 심장 및 폐기능이 약해져서 수명이 단축될 수도 있다. 고혈압, 당뇨병, 고콜레스테롤혈증, 동맥경화증 등의 복합적인 요인들이 심장기능에 압박을 주어 심부전증에 빠지고 돌발사를 유발하기도 한다. 여성의 경우는 여기에 더해 자궁내막암의 발병률이 높아지며 담석증이 생길 가능성도 높아진다. 남성에게는 대장암과 전립선암의 발병확률이 높아진다. 뿐만 아니라, 체중이 증가하면 남들이 자신을 바라보는 시선을 의식하지않을 수 없고 이에 대한 강박관념이 심해지면 사람들을 기피하고 혼자만 지내려는 증상이 생긴다. 또 신체에서 오는 열등감이 자신의 삶에 대한 자신감을 잃게 하여 자아에까지 영향을 미친다.As such, obesity causes various adult diseases. Increased weight can lead to diabetes and lower blood sugar, which can lead to hypertension. If the body weight is 20% higher than normal weight, the likelihood of developing diabetes increases to 10 times that of normal people. In addition, cardiovascular and pulmonary function may be impaired and life expectancy may be shortened. Complex factors such as hypertension, diabetes, hypercholesterolemia, and atherosclerosis may put pressure on the heart function and lead to heart failure and sudden death. In addition to women, the incidence of endometrial cancer is high, and cholelithiasis is more likely to occur. Men are more likely to develop colon cancer and prostate cancer. In addition, as the weight increases, people can not help but be conscious of the gaze that they are looking at, and if the obsession about it becomes worse, the symptoms of avoiding people and living alone are generated. The inferiority feeling that comes from the body also affects the self by losing confidence in his life.
이러한 비만을 예방하고 건강 체중을 유지하기 위해서는 균형잡힌 식사와 규칙적인 운동으로 평생동안 꾸준히 관리해야 한다. 특히 지방세포는 한번 만들어지면 크기는 줄일 수 있지만 자연적으로 제거하는 것은 불가능하여 영구적으로 인체에 남아있게 되므로 처음부터 비만이 되지 않도록 예방하는 것이 중요하다. 따라서, 일차적으로는 섭취하는 칼로리의 양을 줄이고 활동 및 운동량을 증가시키는 방법으로 체중을 줄일 수 있다.In order to prevent obesity and maintain a healthy weight, balanced diet and regular exercise should be maintained throughout the lifetime. In particular, fat cells can be reduced in size once they are made, but it is impossible to remove them naturally and it is left permanently in the human body, so it is important to prevent obesity from beginning. Thus, primarily, you can reduce your weight by reducing the amount of calories consumed and increasing activity and exercise.
그러나 상기한 바와 같이 일단 형성된 지방세포는 제거가 불가능하므로, 다어트 식이 섭취와 더불어 지방흡수를 감소시키는 등의 약물을 복용하는 경우가 있다. 그러나, 이러한 다이어트 약은 고혈압 등의 심혈관계질환 및 뇌졸중과 같은 부작용을 유발할 수 있으며, 복용을 중단할 경우 다시 체중이 증가하는 경우가 빈번하다. 중증 비만의 경우 수술요법을 시행할 수 있으며, 이러한 수술요법으로는 지방을 제거하는 수술이나 몸이 소화할 수 있는 음식량을 제한하기 위한 위성형술 또는 위밴드삽입술 등이 시행되고 있다.
However, as described above, since the fat cells once formed can not be removed, there are cases where a drug such as a daurate diet is consumed along with a decrease in fat absorption. However, such diet drugs can cause side effects such as cardiovascular diseases and strokes such as hypertension, and the weight is frequently increased again when the administration is stopped. In the case of severe obesity, surgical treatment can be performed. Such surgery is performed by surgery to remove fat or gastroplasty or gastric banding to limit the amount of food that can be digested by the body.
이에 본 발명자들은 항비만 효과를 나타내면서 부작용이 적은 천연물을 찾고자 예의 연구노력한 결과, 한약재 백두구 추출물이 지방세포로의 분화를 억제하여 중성지방 생성율을 감소시키는 것을 확인하고 본 발명을 완성하였다.
Accordingly, the inventors of the present invention have made efforts to search for natural products having anti-obesity effects and less side effects, and as a result, they have confirmed that the extract of Baekduguchu can inhibit differentiation into adipocytes and reduce the triglyceride production rate.
본 발명의 하나의 목적은 백두구 추출물을 유효성분으로 포함하는 비만 예방 또는 치료용 약학 조성물을 제공하는 것이다.It is an object of the present invention to provide a pharmaceutical composition for preventing or treating obesity, which comprises an extract of Baekdugi as an active ingredient.
본 발명의 다른 목적은 백두구 추출물을 유효성분으로 포함하는 비만 예방 또는 개선용 건강기능식품을 제공하는 것이다.
Another object of the present invention is to provide a health functional food for preventing or ameliorating obesity comprising Baekdugu extract as an active ingredient.
상기 목적을 달성하기 위한 하나의 양태로서, 본 발명은 백두구 추출물을 유효성분으로 포함하는 비만 예방 또는 치료용 약학 조성물을 제공한다.In one aspect of the present invention, the present invention provides a pharmaceutical composition for preventing or treating obesity, which comprises an extract of Baekduguk as an active ingredient.
본 발명의 용어 "비만(obesity)"은 체지방이 과도한 상태를 말하는 것으로, 임상적으로 체질량지수가 한국의 경우 25, 세계보건기구(WHO)에 의하면 30 이상인 경우를 의미한다. 이때 체질량지수(body mass index; BMI)는 신장의 제곱에 대한 체중(kg/m2)으로 계산된다. 일반적으로 체중이 정상치보다 높은 경우를 의미하지만 체중이 많이 나가지 않더라도 몸의 구성성분 중 체지방의 비율이 높은 경우 비만이라고 진단하며, 성인과 어린이 모두에서 발병하는 질환이다. 비만은 생명을 단축시키며, 고혈압, 당뇨, 심장질환 및 일부 암과 같은 질환에 걸릴 위험에 처하게 할 수 있다. 비만인 경우 건강에 문제가 생길 확률이 높으며 비만의 정도가 증가할수록 위험성 또한 높아진다. 비만이 생기는 이유는 매우 다양하며 여러 가지 원인이 복합적으로 작용하는 경우가 많다. 이 중 가장 빈번한 원인은 유전적 영향, 생리적 영향, 음식섭취 및 섭식장애, 생활방식, 체중력, 임신 또는 약물 등이다. 비만의 첫번째 징후는 평균을 웃도는 체중이상이며 증상은 하기와 같다: 수면문제, 수면무호흡, 호흡짧아짐, 정맥류, 피부문제, 담석 또는 체중부하로 인한 골관절염 등. 비만은 또한 고혈압을 비롯한 고혈당(당뇨), 고콜레스테롤혈증, 고중성지방혈증 등의 질환과 관련이 있다. 한번 과체중이 되면 그 상태를 벗어나기가 쉽지 않으므로 비만은 대부분 평생 겪게 된다. 하지만 비만에 의해 생긴 질병이나 문제는 체중이 감소되면 호전될 수 있다.The term "obesity " of the present invention refers to an excessive state of body fat, which means that the body mass index is clinically 25 in the case of Korea and 30 or more according to the World Health Organization (WHO). The body mass index (BMI) is calculated as body weight (kg / m 2 ) relative to the height of the kidney. Generally, it means that the body weight is higher than the normal value. However, when the body fat percentage of the body composition is high, even if the body weight does not go out, it is diagnosed as obesity. Obesity can shorten life and put you at risk for diseases like hypertension, diabetes, heart disease and some cancers. Obesity is likely to cause health problems, and as the degree of obesity increases, the risk also increases. Obesity is caused by a wide variety of reasons and many causes are often combined. The most frequent causes are genetic influences, physiological effects, food intake and eating disorders, lifestyle, weight, pregnancy or drugs. The first signs of obesity are above average weight and symptoms are as follows: sleep problems, sleep apnea, shortness of breath, varicose veins, skin problems, osteoarthritis due to gallstones or weight loss, Obesity is also associated with diseases such as hypertension, hyperglycemia (diabetes), hypercholesterolemia, and hypertriglyceridemia. Once you are overweight, it is not easy to get out of it, so obesity is a lifelong experience. However, diseases or problems caused by obesity can improve if weight is reduced.
이하 본 발명에서는 "항비만(anti-obesity)"라는 용어를 비만 예방, 치료 또는 개선을 모두 포함하는 의미로 사용한다. 이에 따라, 본 발명에서 사용되는 "항비만 조성물"이라는 용어는 상기 비만 예방 또는 치료용 약학 조성물을 의미한다.In the present invention, the term "anti-obesity " is used to mean prevention, treatment, or improvement of obesity. Accordingly, the term "anti-obesity composition" used in the present invention means a pharmaceutical composition for preventing or treating obesity.
본 발명에서, 용어 "예방"이란 본 발명에 따른 약학적 조성물의 투여에 의해 비만의 발병을 억제 또는 지연시키는 모든 행위를 의미하고, "치료"란 상기 약학적 조성물의 투여에 의해 비만의 의심 및 발병 개체의 증상이 호전되거나 이롭게 변경하는 모든 행위를 의미한다.In the present invention, the term "prevention" means any action which inhibits or delays the onset of obesity by the administration of the pharmaceutical composition according to the present invention, and "treatment" Means any action that alters or alleviates symptoms of an onset individual.
본 발명의 용어 "백두구"는 생강과에 속하는 다년생 초본인 백두구와 자바백두구의 과실을 지칭하는 것으로, 다골(多骨), 백구(白寇), 각박(殼泊)이라고도 불리며, 아모멈 카르다모멈 린네(Amomum cardamomum Linne)라는 학명을 갖는다. 주로 성숙한 과실의 과피를 제거한 것을 백두구라고 한다. 상기 백두구의 과실은 길이 1 내지 2 cm, 지름 5 내지 10 mm 크기로 거의 원구형을 이룬다. 바깥면은 엷은 황백색 내지 황갈색이고 3줄의 둔한 능과 여러 개의 세로줄이 있으며, 윗쪽은 움푹 들어갔고 아랫쪽은 과병이 붙었던 흔적이 있다. 가을에 황녹색을 띤 성숙한 과실을 채취하여 과병과 과피를 제거하고 햇빛에 말려 사용한다. 백두구는 캄보디아, 베트남, 태국이 주산지이며, 자바백두구는 인도네시아가 주산지이나, 상기 2종 모두 중국의 남부지방에서 재배한다. 소화기계 질환에 자주 쓰이며, 몸안의 습을 없애고 몸을 따뜻하게 하여 기를 잘 통하게 하고 구토를 멈추는 효과가 있는 것으로 알려져 있다. 또한 위액분비를 촉진하고 위장의 운동을 강화하여 장내 이상발효를 억제하고 가스를 잘 나가게 하여 입맛을 돋구고 소화불량 및 식적 등을 없앤다. 만성위염으로 인한 위통이나 트림, 구토 등의 증상에 좋으며, 술독을 제거하는 작용이 있다. 그러나, 아직까지 상기 백두구의 항비만 활성에 대해서는 보고된 바 없다.The term "Baekdugugu" of the present invention refers to the fruit of Baekdugu and Java Baekdu, which are perennial herbs belonging to the ginger family, and is also called polybole, white pheasant, chestnut, It has the scientific name Amomum cardamomum Linne. Mainly of mature fruit The removal of the peel is called Baekdugu. The fruit of Baekduju is about 1 to 2 cm in length and 5 to 10 mm in diameter and has a nearly circular shape. Outer surface is pale yellowish white to yellowish brown, with three rows of dull facies and several vertical lines, with a depression at the top and a bruise at the bottom. In the fall, mature fruit with yellowish green color is picked up to remove the disease and the skin and dried in the sun. Baekdugu is the main production area of Cambodia, Vietnam and Thailand. Java Baekdu is the main production area of Indonesia, but both of these are cultivated in the southern part of China. It is often used for digestive system diseases, It is said to have the effect of stopping the vomiting by letting the body warm by the body. It also promotes gastric secretion and strengthens the gastrointestinal motility, suppressing abnormal fermentation in the intestines, promoting gas well, improving taste, and eliminating digestive and gastrointestinal problems. It is good for symptoms such as stomach pains, trims and vomiting caused by chronic gastritis, and it has an action to remove the digest. However, the anti-obesity activity of Baekdudu has not been reported yet.
본 발명의 용어 "추출물(extract)"은 생약을 적절한 침출액으로 짜내고 침출액을 증발시켜 농축한 제제를 의미하는 것으로, 이에 제한되지는 않으나, 추출처리에 의해 얻어지는 추출액, 추출액의 희석액 또는 농축액, 추출액을 건조하여 얻어지는 건조물, 이들의 조정제물 또는 정제물일 수 있다. 상기 백두구 추출물은 당업계에 공지된 일반적인 추출방법, 분리 및 정제방법을 이용하여 제조할 수 있다. 상기 추출방법으로는, 이에 제한되지는 않으나, 바람직하게 열탕 추출, 열수 추출, 냉침 추출, 환류 냉각 추출 또는 초음파 추출 등의 방법을 사용할 수 있다.The term "extract " of the present invention means a preparation which is obtained by squeezing a herbal medicine with an appropriate leaching solution and concentrating by evaporating the leaching solution, and is not limited thereto. The extract, the diluted solution, A dried product obtained by drying, a controlled preparation thereof, or a purified product thereof. The Baekdugu extract can be prepared by using common extraction, isolation and purification methods known in the art. The extraction method may be, but not limited to, hot water extraction, hot water extraction, cold extraction, reflux cooling extraction, or ultrasonic extraction.
본 발명의 구체적인 실시예에 의하면, 본 발명의 백두구 추출물은 일정 중량의 백두구를 분말로 파쇄하고 5 내지 10배의 용매를 가하여 추출하였다. 용매로는 유기용매를 사용할 수 있으며, 상기 유기용매는 C1 내지 C4의 저급 알콜 또는 이들의 혼합용매일 수 있고, 바람직하게는 메탄올일 수 있다. 또한 상기 추출은 바람직하게 65 내지 85℃의 온도에서 수행될 수 있으며, 보다 바람직하게 75℃에서 수행될 수 있다. 상기 추출은 수회 반복하여 수행되었으며, 추출한 시료는 여과 후 감압농축기로 농축하고 동결건조하였다.According to a specific embodiment of the present invention, the Baekdugu extract of the present invention is prepared by crushing Baekdudugu of a predetermined weight into powder and extracting it with 5 to 10 times of a solvent. As the solvent, an organic solvent may be used, and the organic solvent may be a C1 to C4 lower alcohol or a mixture thereof, preferably methanol. Further, the extraction can be preferably carried out at a temperature of 65 to 85 캜, and more preferably at 75 캜. The extraction was repeated several times, and the extracted sample was filtered, concentrated by a vacuum concentrator, and lyophilized.
본 발명에서 상기 비만 예방 또는 치료는 지방전구세포의 지방세포로의 분화억제에 의해 달성되는 것일 수 있다. 본 발명의 백두구 추출물은 지방형성에 관여하는 핵심인자인 C/EBPα, C/EBPβ 및 PPARγ의 발현을 감소시킴으로써 지방세포 생성을 억제하는 활성을 나타낸다.In the present invention, the prevention or treatment of obesity may be achieved by inhibiting differentiation of adipose precursor cells into adipocytes. The extract of Baekduguchi of the present invention shows an activity of inhibiting adipocyte formation by decreasing the expression of C / EBP?, C / EBP? And PPAR? Which are key factors involved in fat formation.
본 발명에서 용어 "C/EBP(CCAAT enhancer binding protein)"란, 염기성-루신 지퍼 모티프(basic-leucine zipper motif)를 가진 전사인자 가운데 하나로, 이들은 지방세포에 한정되어 발현되지는 않으며, 과립구(granulocyte)나 간세포에서도 발현하여 상기 세포들의 최종분화에 중요한 기능을 담당하는 것으로 알려져 있다. 또한, "PPAR-γ(peroxisome proliferator activated receptor-γ)"란, PPARs(peroxisome proliferator activated receptors)인 핵수용체(nuclear hormone receptor) 수퍼 패밀리(superfamily)에 속하는 것으로, PPARα, PPARγ 및 PPARδ 등이 상기 수퍼패밀리에 포함되며 지방세포 분화 조절 기능을 하는 것으로 알려져 있다.In the present invention, the term "C / EBP (CCAAT enhancer binding protein)" is one of the transcription factors having a basic-leucine zipper motif, which is not expressed in adipocytes but granulocyte ) Or hepatocytes, and is known to play an important role in the final differentiation of the cells. PPAR? (Peroxisome proliferator activated receptor-?) Belongs to a nuclear hormone receptor superfamily which is peroxisome proliferator activated receptors (PPARs), and PPAR ?, PPAR? And PPAR? Family, and is known to regulate adipocyte differentiation.
상기 전사인자들은 지방전구세포(preadipocyte)가 증식 및 분화되는 과정을 거쳐 성숙한 지방세포(adipocyte)로 되는 과정인 지방세포분화(Adipogenesis)에 관여하는 일련의 유전자 발현을 조절하는 역할을 하는데, 특히, PPARγ와 C/EBP 패밀리는 호르몬에 의해 지방세포분화가 유도되는 과정에서 중요하게 작용하는 전사인자로, 지방세포분화를 총괄적으로 조절하는 역할을 하며, 지방세포로 분화된 상태를 유지하는데 필수적인 인자이다. 또한, C/EBPβ는 PPARγ발현을 촉진시켜 지방전구세포의 초기 분화과정에서 중요한 역할을 하는 한편, PPARγ와의 강한 상승작용을 통해 지방전구세포의 말기 분화과정을 촉진하는 역할을 한다.The transcription factors regulate the expression of a series of genes involved in adipogenesis, which is a process of becoming a mature adipocyte through the process of proliferation and differentiation of a preadipocyte, The PPARγ and C / EBP family are important transcription factors that are involved in the induction of adipocyte differentiation by hormones. They play a role in regulating adipocyte differentiation and are essential factors to maintain differentiation into adipocytes . In addition, C / EBPβ plays an important role in the early differentiation process of lipoprotein by promoting PPARγ expression and promotes the late differentiation process of lipid precursor cells through strong synergistic action with PPARγ.
본 발명의 항비만 조성물은 체중감소, 체지방감소, 혈중 중성지방 함량 감소 및 혈중 콜레스테롤 함량 감소의 효과를 가진다. 본 발명의 구체적인 실시예에 의하면, IBMX(3-isobutyl-1-methylxanthine), 덱사메타손 및 인슐린을 포함하는 지방세포 분화배지를 이용하여 3T3-L1 지방전구세포의 분화를 유도한 경우, 상기 본 발명의 백두구 추출물을 첨가하여 지방세포 생성을 억제함으로 추출물의 농도에 따라 중성지방 함량이 감소하는 것을 확인할 수 있었다(도 3 및 도 5).
The anti-obesity composition of the present invention has effects of weight loss, decrease of body fat, decrease of blood triglyceride content and decrease of blood cholesterol content. According to a specific embodiment of the present invention, when differentiation of 3T3-L1 adipose precursor cells is induced using an adipogenic differentiation medium containing IBMX (3-isobutyl-1-methylxanthine), dexamethasone and insulin, It was confirmed that the content of triglyceride was decreased according to the concentration of the extract by inhibiting adipocyte production by adding Baekduguchi extract (Figs. 3 and 5).
추가적으로 상기 본 발명의 항비만 조성물은 지방생성을 억제함으로써, 비만 뿐만 아니라 비만과 직·간접적으로 관련된 지방간 또는 지질관련 심혈관 질환에 대한 예방 또는 치료의 효과를 나타낼 수 있다.In addition, the anti-obesity composition of the present invention can prevent or treat obesity as well as fatty liver or lipid-related cardiovascular diseases directly or indirectly associated with obesity by inhibiting lipogenesis.
본 발명의 용어 "지방간"은 지방질, 그 중에서도 특히 중성지방(triglyceride)이 간세포에 축정되어 간 무게의 5% 이상을 차지하고 있는 질병이다. 음식물 등을 통하여 섭취한 지방질을 원활하게 처리하지 못하는 경우 발생하기도 하지만, 지방간을 일으키는 대표적인 원인은 술이다. 술은 간의 여러 가지 대사기능, 특히 지방산 산화 분해력을 감소시켜 간에 지방을 축적시킨다. 비만한 사람에게도 자주 지방간이 발생하며, 그 외, 당뇨병, 갑상선 기능 항진증 등의 내분비 질환, 스테로이드 호르몬의 과다 사용이나 심한 영양부족에 의해서도 발생할 수 있다. 특별한 증상이 없으므로 증상만으로 진단할 수 없으며, 간 질환의 일반적인 증상인 피로감, 식욕부진, 무기력 등이 나타날 수 있고, 오른쪽 갈비뼈 아래쪽에 불쾌감이나 둔통을 느끼기도 한다.The term " fatty liver "of the present invention refers to a disease in which fat, particularly triglyceride, is localized in hepatocytes and accounts for more than 5% of liver weight. However, the most common cause of fatty liver disease is alcohol. Alcohol reduces various metabolic functions of the liver, especially fatty acid oxidative degradation, and accumulates fat in the liver. Fatty liver often occurs in obese people, and it can also be caused by endocrine diseases such as diabetes, hyperthyroidism, overuse of steroid hormone, or severe nutritional deficiency. There are no special symptoms, so it is not possible to diagnose it by symptoms alone. Fatigue, anorexia and lethargy, which are common symptoms of liver disease, can occur and feel discomfort or dullness in the lower part of the right rib.
본 발명의 용어 "지질관련 심혈관 질환"은 지방산이나 콜레스테롤 등이 과도한 섭취나 대사 장애로 인해 축적되어 발생하는 질병이다. 특히 혈액 중의 중성지방의 농도가 비정상적으로 높아지는 고중성지방혈증은 미국 심장협회에 의해 심혈관질환의 위험인자로 규정되어 있다. 혈장중성지방은 식후 급격히 증가하여 약 8시간 후에는 식전수준으로 돌아온다. 중성지방은 당뇨병, 비만, 음주자에서 증가하며 당분을 많이 섭취하는 사람에서도 증가한다. 그 외 갑상선 기능저하, 신증후군, 신전증, 피임약 및 대량의 이뇨제와 베타차단제도 중성지방을 증가시킬 수 있다. 혈중중성지방이 증가할수록 LDL(low density lipoprotein)-콜레스테롤이 증가하며, 상기 LDL은 죽상경화를 촉진시킨다. 상기 지질관련 심혈관 질환으로는 고지혈증, 심장병, 고혈압, 동맥경화증 등이 포함되나, 이에 제한되지 않는다.The term "lipid-related cardiovascular disease" of the present invention is a disease caused by accumulation of fatty acid or cholesterol due to excessive intake or metabolic disorder. Hypertriglyceridemia, in which the concentration of triglycerides in the blood is abnormally high, is defined by the American Heart Association as a risk factor for cardiovascular disease. Plasma triglyceride increases rapidly after meals and returns to the pre-meal level after about 8 hours. Triglycerides increase in diabetes, obesity, and drinkers, and increase in people who consume a lot of sugar. Other hypothyroidism, nephrotic syndrome, hypertension, birth control pills, and large amounts of diuretics and beta blockers can also increase triglycerides. As the blood triglyceride increases, LDL (low density lipoprotein) - cholesterol increases and the LDL promotes atherosclerosis. The lipid-related cardiovascular diseases include, but are not limited to, hyperlipidemia, heart disease, hypertension, arteriosclerosis and the like.
상기 본 발명의 약학조성물은 약학적으로 허용가능한 담체를 추가로 포함할 수 있다. 본 발명의 용어 "약학적으로 허용가능한"이란 상기 조성물에 노출되는 세포나 인간에게 독성이 없는 특성을 나타내는 것을 의미한다. 상기 담체는 완충제, 보존제, 무통화제, 가용화제, 등장제, 안정화제, 기제, 부형제, 윤활제 등 당업계에 공지된 것이라면 제한없이 사용할 수 있다. 또한 본 발명의 약학조성물은 각각 통상의 방법에 따라 산제, 과립제, 정제, 캡슐제, 현탁액, 에멀젼, 시럽, 에어로졸 등의 경구형 제형, 외용제, 좌제 및 멸균 주사용액의 형태로 제형화하여 사용될 수 있다. 나아가, 연고제, 로션제, 스프레이제, 패취제, 크림제, 산제, 현탁제, 겔제 또는 젤의 형태의 피부 외용제의 형태로 사용될 수 있다. 본 발명의 조성물에 포함될 수 있는 담체, 부형제 및 희석제로는 락토즈, 덱스트로즈, 수크로스, 솔비톨, 만니톨, 자일리톨, 에리스리톨, 말티톨, 전분, 아카시아 고무, 알지네이트, 젤라틴, 칼슘 포스페이트, 칼슘 실리케이트, 셀룰로즈, 메틸 셀룰로즈, 미정질 셀룰로스, 폴리비닐 피롤리돈, 물, 메틸히드록시벤조에이트, 프로필히드록시벤조에이트, 탈크, 마그네슘 스테아레이트 및 광물유를 들 수 있다. 제제화할 경우에는 보통 사용하는 충진제, 증량제, 결합제, 습윤제, 붕해제, 계면활성제 등의 희석제 또는 부형제를 사용하여 조제된다. 경구투여를 위한 고형제제에는 정제, 환제, 산제, 과립제, 캡슐제 등이 포함되며, 이러한 고형제제는 상기 백두구 추출물에 적어도 하나 이상의 부형제 예를 들면, 전분, 칼슘카보네이트 (calcium carbonate), 수크로스(sucrose) 또는 락토오스(lactose), 젤라틴 등을 섞어 조제된다. 또한 단순한 부형제 이외에 마그네슘 스티레이트, 탈크 같은 윤활제들도 사용된다. 경구를 위한 액상 제제로는 현탁제, 내용액제, 유제, 시럽제 등이 해당되는 데 흔히 사용되는 단순희석제인 물, 리퀴드 파라핀 이외에 여러 가지 부형제, 예를 들면 습윤제, 감미제, 방향제, 보존제 등이 포함될 수 있다. 비경구 투여를 위한 제제에는 멸균된 수용액, 비수성용제, 현탁제, 유제, 동결건조 제제, 좌제가 포함된다. 비수성용제, 현탁제로는 프로필렌글리콜 (propylene glycol), 폴리에틸렌 글리콜, 올리브오일과 같은 식물성 기름, 에틸올레이트와 같은 주사 가능한 에스테르 등이 사용될 수 있다. 좌제의 기제로는 위텝솔(witepsol), 마크로골, 트윈 (tween) 61, 카카오지, 라우린지, 글리세로제라틴 등이 사용될 수 있다.The pharmaceutical composition of the present invention may further comprise a pharmaceutically acceptable carrier. The term "pharmaceutically acceptable" of the present invention means that it exhibits properties that are not toxic to the cells or humans exposed to the composition. Such carriers may be used without limitation as long as they are known in the art such as buffers, preservatives, wetting agents, solubilizers, isotonic agents, stabilizers, bases, excipients and lubricants. In addition, the pharmaceutical composition of the present invention may be formulated in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols and the like, oral preparations, suppositories and sterilized injection solutions according to a conventional method have. Furthermore, it can be used in the form of an external preparation for skin in the form of ointments, lotions, spray agents, patches, creams, powders, suspensions, gels or gels. Examples of carriers, excipients and diluents that can be included in the composition of the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, Cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. In the case of formulation, a diluent or excipient such as a filler, an extender, a binder, a wetting agent, a disintegrant, or a surfactant is usually used. Solid formulations for oral administration include tablets, pills, powders, granules, capsules and the like, which may contain at least one excipient such as starch, calcium carbonate, sucrose sucrose), lactose, gelatin, and the like. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Liquid preparations for oral use may include various excipients such as wetting agents, sweetening agents, fragrances, preservatives, etc. in addition to water and liquid paraffin, which are simple diluents commonly used in suspension agents, solutions, emulsions and syrups have. Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Examples of the suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like. Examples of the suppository base include witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatin and the like.
한편, 본 발명의 약학조성물은 약학적으로 유효한 양으로 투여한다. 본 발명의 용어 "투여"란, 적절한 방법으로 개체에게 소정의 물질을 도입하는 것을 의미하며 상기 조성물의 투여 경로는 목적 조직에 도달할 수 있는 한 어떠한 일반적인 경로를 통하여 투여될 수 있다. 복강내 투여, 정맥내 투여, 근육내 투여, 피하 투여, 피내 투여, 경구 투여, 국소 투여, 비내 투여, 폐내 투여, 직장내 투여될 수 있으나, 이에 제한되지는 않는다.Meanwhile, the pharmaceutical composition of the present invention is administered in a pharmaceutically effective amount. The term "administering" of the present invention means introducing a predetermined substance into an individual by an appropriate method, and the administration route of the composition can be administered through any conventional route so long as it can reach the target tissue. But are not limited to, intraperitoneal, intravenous, intramuscular, subcutaneous, intradermal, oral, topical, intranasal, intrathecal, rectal.
상기 용어 "개체"란 비만이 발병하였거나 발병할 수 있는 인간을 포함한 쥐, 생쥐, 가축 등의 모든 동물을 의미한다. 바람직하게는, 인간을 포함한 포유동물일 수 있다.The term "individual " refers to all animals, including mice, mice, livestock, etc., including humans who have developed or are capable of developing obesity. Preferably, it may be a mammal, including a human.
상기 용어 "약학적으로 유효한 양"이란 의학적 치료에 적용 가능한 합리적인 수혜/위험 비율로 질환을 치료하기에 충분하며 부작용을 일으키지 않을 정도의 양을 의미하며, 유효 용량 수준은 환자의 성별, 연령, 체중, 건강상태, 질병의 종류, 중증도, 약물의 활성, 약물에 대한 민감도, 투여 방법, 투여 시간, 투여 경로, 및 배출 비율, 치료 기간, 배합 또는 동시에 사용되는 약물을 포함한 요소 및 기타 의학 분야에 잘 알려진 요소에 따라 당업자에 의해 용이하게 결정될 수 있다. 바람직하게 본 발명의 백두구 추출물은 고형분을 기준으로 1일 0.0001 내지 100 mg/체중 kg으로, 바람직하게는 0.001 내지 100 mg/체중 kg으로 투여할 수 있다. 투여는 상기 권장 투여량을 하루에 한번 투여할 수도 있고, 수회 나누어 투여할 수도 있다.The term "pharmaceutically effective amount" means an amount sufficient to treat a disease at a reasonable benefit / risk ratio applicable to medical treatment and not causing side effects, and the effective dose level is determined by the patient's sex, Including, but not limited to, medicaments and other medical fields that are used in combination, or in combination with, or in combination with, a pharmaceutically acceptable carrier, excipient, Can be readily determined by those skilled in the art according to known factors. Preferably, the Baekdugu extract of the present invention may be administered at a daily dose of 0.0001 to 100 mg / kg of body weight, preferably 0.001 to 100 mg / kg of body weight, based on the solid content. The administration may be carried out once per day, or divided into several doses.
따라서, 다른 하나의 양태로서 본 발명은 상기 백두구 추출물을 유효성분으로 포함하는 약학 조성물을 개체에 투여하는 단계를 포함하는 비만의 치료방법을 제공할 수 있다.Accordingly, in another aspect, the present invention provides a method for treating obesity comprising the step of administering to a subject a pharmaceutical composition comprising the Baekdugu extract as an active ingredient.
본 발명에서, 용어 "개체"란 상기한 바와 같이 비만이 발병하였거나 발병할 수 있는 인간을 포함한 모든 동물을 의미하고, 본 발명의 약학적 조성물을 개체에게 투여함으로써 상기 비만을 효과적으로 예방 또는 치료할 수 있다. 본 발명의 약학적 조성물은 기존의 비만 치료제와 병행하여 투여될 수 있다.In the present invention, the term "individual" means all animals, including humans, who have developed or are capable of developing obesity as described above, and the obesity can be effectively prevented or treated by administering the pharmaceutical composition of the present invention to a subject . The pharmaceutical composition of the present invention can be administered in parallel with the existing therapeutic agent for obesity.
본 발명에 따른 약학 조성물은 희렴(Siegesbeckia pubescens) 추출물을 추가로 포함할 수 있다.The pharmaceutical compositions according to the invention huiryeom (Siegesbeckia pubescens ) extracts.
상기 본 발명의 용어 "희렴"은 희첨, 시첨, 저고 또는 진득찰이라고도 불리는 국화과 식물 또는 이의 전초를 건조시킨 한약재를 지칭하는 것으로, "희렴 추출물"은 상기 희렴으로부터 물 또는 C1 내지 C4의 저급 알콜을 이용하여 추출한 추출물일 수 있다. 상기 희렴은 풀의 냄새가 돼지와 비슷하고 맵고 쏘는 맛이 있다. 호랑이나 개에게 물린 상처를 치료할 때 사용된다 하여 호고 또는 구고라고 불리기도 한다. 동속의 식물로 진득찰속에 진득찰(Sigesbeckia glabrescens), 제주진득찰(Siegebeckia orientalis L.), 털진득찰(Siegesbeckia pubescens)로 나뉘어지는데, 이중 털진득찰은 진득찰에 비하여 줄기와 잎에 털이 많고 꽃자루에 선이 있는 것이 특징이고, 줄기와 잎뒤의 맥상에 백색의 긴 털이 밀생(密生)을 하고 잎이 대형(大形)이며 화편(花梗)에 흔히 선모(腺毛)가 있고 수과(瘦果)는 2.5∼3.5mm로 보다 길다. 전통적으로 풍습을 제거해 관절염, 사지동통마비, 하지무력, 중풍으로 인한 반신불수 등에 쓰고, 이외에 종기, 발진, 피부가려움증, 습진 등에 쓴다. 또한 고혈압, 두통, 어지럼증, 급성간염 등에 사용되며, 혈관확장작용이 있어 혈압강하작용을 나타내는 것으로 알려져 있어 고혈압 환자는 차로 복용할 수 있다. 한편 본 발명자들은 선행연구로부터 희렴 추출물이 상기한 효능 이외에 지방생성을 억제하는 항비만 효과를 나타냄을 확인하였으며, 이를 대한민국 특허출원 제10-2013-0023603호로 출원한 바 있다.The term " pleiotropic " of the present invention refers to a herbaceous herb that has dried a herbaceous plant or its outpost, which is also referred to as a scavenger, And the like. The flavor of this glue is similar to that of pigs and has a spicy taste. It is sometimes used to treat bruises that are touched by tigers or dogs. It is divided into Sigebeckia glabrescens , Siegebeckia orientalis L. , and Siegesbeckia pubescens . It has a hairy stem and a leaf on the stem and a line on the peduncle. White long hairs are densely grown in stem and leaf vein, and leaf is large in shape. There are often glandular hair on petiole and 2.5 ~ 3.5mm in diameter. Is longer. Traditionally, it is used to remove arthritis, limb paralysis, leg weakness, paralysis due to paralysis, and other symptoms such as swelling, rash, skin itching, and eczema. It is also used for hypertension, headache, dizziness, acute hepatitis, and is known to exhibit blood pressure lowering effect due to vasodilating action, so patients with hypertension can take it by car. On the other hand, the inventors of the present invention have confirmed from the preceding studies that the extract of the present invention exhibits an anti-obesity effect that inhibits lipogenesis in addition to the above-mentioned efficacy, and has filed this application in Korean Patent Application No. 10-2013-0023603.
본 발명의 구체적인 실시예에 의하면, 본 발명의 백두구 추출물과 동일한 방법으로 제조한 희렴 추출물을 1:1로 혼합한 혼합물이 동일한 농도로 사용된 개별 추출물에 비해 높은 항비만 효과를 나타냄을 확인하였다(도 7).
According to a specific example of the present invention, it was confirmed that a mixture of 1: 1 mixture of the extract of the present invention and the mixture of the extract prepared by the same method as the Baekdugu extract of the present invention showed a higher anti-obesity effect than the individual extracts used at the same concentration 7).
다른 하나의 양태로서, 본 발명은 백두구 추출물을 유효성분으로 포함하는 비만 예방 또는 개선용 건강기능식품을 제공한다.In another aspect, the present invention provides a health functional food for preventing or improving obesity comprising Baekdugu extract as an active ingredient.
또한 본 발명에 따른 건강기능식품은 추가로 희렴 추출물을 포함하는 것일 수 있다.In addition, the health functional food according to the present invention may further comprise a flavor extract.
상기 백두구 및 희렴 추출물에 대해서는 전술한 바와 같다.The Baekduk and the Rice extract are as described above.
본 발명의 조성물을 식품 첨가물로 사용할 경우, 상기 백두구 추출물을 그대로 첨가하거나 다른 식품 또는 식품 성분과 함께 사용할 수 있고, 통상의 방법에 따라 적절하게 사용할 수 있다. 유효 성분의 혼합양은 사용 목적(예방, 건강 또는 치료적 처치)에 따라 적합하게 결정될 수 있다.When the composition of the present invention is used as a food additive, the Baekdugu extract may be added as it is or may be used together with other food or food ingredients, and may be suitably used according to a conventional method. The amount of the active ingredient to be mixed can be suitably determined according to the intended use (prevention, health or therapeutic treatment).
본 발명의 용어 "건강기능식품"은 건강보조의 목적으로 특정성분을 원료로하거나 식품 원료에 들어있는 특정성분을 추출, 농축, 정제, 혼합 등의 방법으로 제조, 가공한 식품을 말하며, 상기 성분에 의해 생체방어, 생체리듬의 조절, 질병의 방지와 회복 등 생체조절기능을 생체에 대하여 충분히 발휘할 수 있도록 설계되고 가공된 식품을 말하는 것으로서, 상기 건강식품용 조성물은 질병의 예방 및 질병의 회복 등과 관련된 기능을 수행할 수 있다.The term "health functional food" of the present invention refers to a food prepared by processing a specific ingredient as a raw material for the purpose of health assisting or by extracting, concentrating, refining, mixing or the like a specific ingredient contained in a food raw material, Refers to foods that are designed and processed so that the body control function such as bio-defense, regulation of bodily rhythm, prevention and recovery of disease can be sufficiently exhibited to the living body, and the composition for health food is used for prevention of diseases, Can perform related functions.
또한, 본 발명의 조성물이 사용될 수 있는 건강식품의 종류에는 제한이 없다. 아울러 본 발명의 백두구 추출물을 활성성분으로 포함하는 조성물은 당업자의 선택에 따라 건강기능식품에 함유될 수 있는 적절한 기타 보조 성분과 공지의 첨가제를 혼합하여 제조할 수 있다. 첨가할 수 있는 식품의 예로는 육류, 소세지, 빵, 쵸코렛, 캔디류, 스낵류, 과자류, 피자, 라면, 기타 면류, 껌류, 아이스크림 류를 포함한 낙농제품, 각종 스프, 음료수, 차, 드링크제, 알콜 음료 및 비타민 복합제 등이 있으며, 본 발명에 따른 추출물을 주성분으로 하여 제조한 즙, 차, 젤리 및 주스 등에 첨가하여 제조할 수 있다.There is no limitation on the kind of health food to which the composition of the present invention can be used. In addition, the composition comprising the Baekdugu extract of the present invention as an active ingredient may be prepared by mixing other suitable auxiliary ingredients, which may be contained in health functional foods, with known additives according to the selection of a person skilled in the art. Examples of foods that can be added include dairy products, such as meat, sausage, bread, chocolates, candies, snacks, confectionery, pizza, ramen, other noodles, gums, ice cream, various soups, drinks, tea, Vitamin complex, and the like, and can be prepared by adding to the juice, tea, jelly, and juice prepared from the extract of the present invention as a main component.
본 발명의 백두구 추출물 또는 이에 희렴 추출물을 추가로 포함하는 혼합물은 천연 한약재를 원료로 하므로 약학적 조성물 또는 식품 첨가물로 사용할 경우에도 일반적인 합성 화합물에 비하여 부작용이 덜할 수 있으므로, 안전하게 약학적 조성물 및 건강기능식품에 포함되어 유용하게 사용될 수 있다.
Since the mixture of Baekdugu extract or the mixture of the present invention further contains a natural medicinal herb as a raw material, even when it is used as a pharmaceutical composition or a food additive, the side effects can be less than general synthetic compounds. Therefore, It can be usefully included in foods.
본 발명의 백두구 추출물을 포함하는 조성물은 지방전구세포의 지방세포로의 분화를 억제하는 능력을 가지므로 항비만 조성물로 유용하게 사용될 수 있으며, 구체적으로 지방세포생성을 억제하므로 지방간, 또는 지질관련 심혈관질환과 같은 불필요한 지방의 축적으로 유발되는 질환의 예방 또는 치료를 위한 약학적 조성물 또는 건강기능식품으로 이용될 수 있다. 또한 항비만 활성을 갖는 희렴 추출물과 혼합하여 시너지적 효과를 나타냄을 확인하였으므로, 희렴 추출물과의 혼합물의 형태로 이용할 수 있다.
The composition of the present invention containing Baekdugu extract can be useful as an anti-obesity composition because it has the ability to inhibit the differentiation of adipose precursor cells into adipocytes. Specifically, the composition inhibits adipocyte production, Or as a health functional food for the prevention or treatment of diseases caused by the accumulation of unnecessary fats such as < RTI ID = 0.0 > Also, it has been confirmed that it has synergistic effect by mixing with the comedium extract having anti-obesity activity, so that it can be used in the form of a mixture with the comedium extract.
도 1은 본 발명의 백두구 추출물의 세포독성을 나타낸 도이다.
도 2는 본 발명의 희렴 추출물의 세포독성을 나타낸 도이다.
도 3은 본 발명의 백두구 추출물의 농도 의존적 지방생성억제를 나타낸 도이다. 백두구 추출물을 각각 표기한 농도로 첨가하여 분화시킨 세포를 오일 레드 오 염색(oil red O staining)하여 (a) 현미경으로 관찰한 결과 및 (b) 상기 염색된 오일 레드 오를 탈색하여 정량함으로써 계산된 중성지방 함량을 도시하였다.
도 4는 본 발명의 희렴 추출물의 농도 의존적 지방생성억제를 나타낸 도이다. 희렴 추출물을 각각 표기한 농도로 첨가하여 분화시킨 세포를 오일 레드 오 염색(oil red O staining)하여 (a) 현미경으로 관찰한 결과 및 (b) 상기 염색된 오일 레드 오를 탈색하여 정량함으로써 계산된 중성지방 함량을 도시하였다.
도 5는 본 발명의 백두구 추출물의 지방형성 유전자 발현에 대한 효과를 나타낸 도이다. 백두구 추출물 농도에 따른 지방형성 관련 유전자의 발현 정도를 RT-PCR을 이용하여 확인하였다.
도 6은 본 발명의 희렴 추출물의 지방형성 유전자 발현에 대한 효과를 나타낸 도이다. 희렴 추출물 농도에 따른 지방형성 관련 유전자의 발현 정도를 RT-PCR을 이용하여 확인하였다.
도 7은 본 발명의 백두구 추출물과 희렴 추출물과의 1:1 혼합물에 의한 시너지적 지방생성억제를 나타낸 도이다. 상기 백두구와 희렴 추출물의 1:1 혼합물을 각각 표기한 농도로 첨가하여 분화시킨 세포를 오일 레드 오 염색(oil red O staining)하여 (a) 현미경으로 관찰한 결과 및 (b) 상기 염색된 오일 레드 오를 탈색하여 정량함으로써 계산된 중성지방 함량을 도시하였다.Brief Description of the Drawings Fig. 1 is a diagram showing cytotoxicity of Baekdugu extract of the present invention. Fig.
FIG. 2 is a graph showing cytotoxicity of the extract of the present invention. FIG.
FIG. 3 is a graph showing the concentration-dependent inhibition of fat production of the extract of Baekduguchi according to the present invention. The results obtained by (a) observing with a microscope the oil-red stained cells obtained by adding Baekdugu extract to the labeled concentrations, and (b) measuring the neutrality The fat content is shown.
4 is a graph showing the concentration-dependent fat generation inhibition of the extract of the present invention. The results are shown in (a) observation under a microscope by oil red o staining and (b) neutralization calculated by discoloring and staining the stained oil red ocher. The fat content is shown.
FIG. 5 is a graph showing the effect of the extract of Paecilomyces sp. On the expression of fat-forming gene of the present invention. The expression level of the lipogenic gene was determined by RT-PCR according to the concentration of Baekdugu extract.
FIG. 6 is a graph showing the effect of the comed extract of the present invention on expression of fat-forming genes. The expression level of the lipogenesis-related gene was determined by RT-PCR.
FIG. 7 is a graph showing inhibition of synergistic lipogenesis by a 1: 1 mixture of Baekduguk extract and Comvitae extract of the present invention. (A) observed under a microscope by oil red o staining, and (b) the results of observation of the dyed oil red The triglyceride content calculated by decolorizing and measuring the broth was shown.
이하, 실시예를 통하여 본 발명의 구성 및 효과를 더욱 상세히 설명하고자 한다. 이들 실시예는 오로지 본 발명을 예시하기 위한 것일 뿐, 본 발명의 범위가 이들 실시예에 의해 한정되는 것은 아니다.
Hereinafter, the constitution and effects of the present invention will be described in more detail through examples. These examples are only for illustrating the present invention, and the scope of the present invention is not limited by these examples.
실시예Example
1: One:
백두구Baekdugu
((
AmomumAmomum
cardamomumcardamomum
L.) 및 L.) and
희렴Celebration
((
SiegesbeckiaSiegesbeckia
pubescenspubescens
) 추출물의 제조) Preparation of extract
백두구와 희렴을 각각 10 g씩 측량하여 분말로 파쇄한 후 메탄올을 이용하여 추출하였다. 시료의 5 내지 10배의 메탄올을 가한 후 75℃에서 추출하였으며, 상기 과정을 3회 반복하여 추출하였다. 추출한 시료는 여과한 후 감압농축기(N-1000S-W, EYELA, Japan)로 농축하고 동결건조(FDU2100, EYELA, Japan)한 후 중량법으로 수율을 계산하였다.Ten grams of Baekduju and 10 kinds of comedones were measured, and then they were pulverized with powder and extracted with methanol. Methanol 5 to 10 times that of the sample was added, and the mixture was extracted at 75 캜. The above procedure was repeated three times. The extracted samples were filtered and concentrated by using a vacuum concentrator (N-1000S-W, EYELA, Japan) and lyophilized (FDU2100, EYELA, Japan).
상기 계산된 백두구 및 희렴 메탄올 추출물의 수율은 각각 2.6%와 18.5%이며 이를 하기 표 1에 나타내었다.The yields of the Baekduk and the methanol extracts were 2.6% and 18.5%, respectively, and are shown in Table 1 below.
실시예Example
2: 백두구 및 2: Baekdugu and
희렴Celebration
추출물의 Extract
항산화능Antioxidant ability
분석 analysis
전자공여능은 항산화 작용의 지표로 사용되고 있으며, 식물 추출물의 항산화능 측정에 널리 사용되고 있다. 전자공여작용은 인체 내에서 생성되는 자유 라디칼의 전자를 공여하여 상기 자유 라디칼에 의한 노화 및 질병을 억제하는 작용을 한다. 상기 실시예 1의 방법으로 제조한 백두구와 희렴 추출물의 전자공여능을 측정하기 위하여 DPPH(2,2-diphenyl-1-picrylhydrazyl) 라디칼 소거 활성(DPPH radical scavenging activity) 측정을 수행하였다. DPPH는 비교적 안정한 자유 라디칼로써, 아스코르브산(ascorbic acid), 토코페롤(tocopherol), 폴리히드록시 방향족 화합물(polyhydroxy aromatic compound) 또는 방향족 아민류(aromatic amines)에 의해 환원되어 짙은 자주색이 탈색되는 원리를 이용하여 흡광도 측정에 의해 간편하게 항산화 활성을 측정할 수 있는 방법이다.Electron donating ability is used as an index of antioxidant activity and it is widely used for measuring the antioxidant ability of plant extracts. The electron donating action serves to donate electrons of free radicals generated in the human body to inhibit aging and diseases caused by the free radicals. DPPH (2,2-diphenyl-1-picrylhydrazyl) radical scavenging activity (DPPH radical scavenging activity) measurement was performed to determine the electron donating ability of Baekdugu and Kurage extract prepared by the method of Example 1 above. DPPH is a relatively stable free radical, which is reduced by ascorbic acid, tocopherol, polyhydroxy aromatic compounds or aromatic amines, and decolorizes the deep purple It is a method which can easily measure antioxidative activity by absorbance measurement.
구체적으로, 백두구 및 희렴 추출물을 각각 농도별(1.25 내지 320 μg/ml)로 메탄올에 녹여 준비한 후, 96 웰 플레이트에 메탄올에 용해된 1.5×10-4 M 농도의 DPPH 40 μl와 다양한 농도별로 준비한 백두구 또는 희렴 추출물 용액 160 μl를 분주한 혼합액을 실온에서 30분간 반응시킨 후, 마이크로플레이트 리더를 이용하여 520 nm에서 흡광도를 측정하였다. 자유 라디칼 소거 활성을 시료를 첨가하지 않은 음성 대조군에 대한 백분율로 나타내고, 50% 저해 농도(IC50)를 계산하였다. 양성 대조군으로는 대표적인 항산화제인 아스코르브산(비타민 C)을 사용하였으며, 측정값은 3회 반복실험하여 평균값으로 나타내었다. 상기 자유 라디칼 소거능의 백분율을 계산하는 공식은 하기와 같다.
Specifically, the Baekdu-gug and the comedium extract were dissolved in methanol at concentrations of 1.25 to 320 μg / ml, respectively. Then, 40 μl of 1.5 × 10 -4 M DPPH dissolved in methanol was added to 96-well plates and prepared at various concentrations The mixed solution, in which 160 μl of Baekdugu or Combi extract solution was dispensed, was reacted at room temperature for 30 minutes, and the absorbance was measured at 520 nm using a microplate reader. The free radical scavenging activity was expressed as a percentage of the negative control to which no sample was added, and the 50% inhibitory concentration (IC 50 ) was calculated. As a positive control, ascorbic acid (vitamin C), a representative antioxidant, was used. The formula for calculating the percentage of the free radical scavenging ability is as follows.
DPPH 라디칼 소거 활성 (%)={1-(A-B)/C}×100
DPPH radical scavenging activity (%) = {1- (AB) / C} 100
상기 A는 520 nm에서 시료의 흡광도, B는 520 nm에서 색 대조군 흡광도, C는 520 nm에서 대조군 흡광도이다.
A is the absorbance of the sample at 520 nm, B is the color control absorbance at 520 nm, and C is the control absorbance at 520 nm.
본 발명의 백두구 및 희렴 메탄올 추출물의 DPPH 라디컬 소거 활성에 대해 계산된 IC50 값을 상기 표 1에 수율과 함께 나타내었다. 백두구 및 희렴 추출물의 DPPH 라디컬 소거 활성에 대해 계산된 IC50 값은 각각 65.23 μg/ml 및 25.15 μg/ml로 양성 대조군으로 사용한 아스코르브산(3.65 μg/ml)에 비해서는 다소 활성이 낮았으나, 단일물이 아닌 추출물임을 감안하면 비교적 높은 항산화능을 보유한 것을 확인할 수 있었다.
The IC 50 values calculated for the DPPH radical scavenging activity of the Baekjugu and comedium methanol extracts of the present invention are shown in Table 1 together with the yield. The IC 50 values calculated for the DPPH radical scavenging activity of Baekdugu and Kwangju extracts were slightly lower than those of ascorbic acid (3.65 μg / ml) used as a positive control at 65.23 μg / ml and 25.15 μg / ml, respectively, Considering that it is an extract that is not a single substance, it has a relatively high antioxidant ability.
실시예Example
3: 백두구 및 3: Baekdugu and
희렴Celebration
추출물의 세포독성 분석 Cytotoxicity analysis of extracts
상기 실시예 1에서 제조한 백두구 및 희렴 추출물의 세포독성을 확인하였다. 구체적으로 마우스 지방전구세포주인 3T3-L1(American Type Culture Collection; ATCC, USA)을 10% 송아지혈청(bovine calf serum; BCS) 및 페니실린/스트렙토마이신(penicillin/streptomycin)이 포함된 DMEM(Dulbecco's Modified Eagle Medium) 배지에서 배양하였다. 24 웰 조직 배양 플레이트에 웰 당 105 개의 세포를 분주하여 24시간 동안 부착시키고 다양한 농도의 백두구(50, 100, 150, 200, 300, 400, 500 μg/ml) 또는 희렴(50, 100, 150, 200, 250, 300, 500 μg/ml) 추출물 용액으로 각각 처리하였다. 72시간 경과 후 시료를 포함하는 배지를 제거하고 WST(water-soluble tetrazolium salt) 시약을 포함하는 신선한 배지로 교체하여 37℃에서 30분간 인큐베이션하고 멀티플레이트 리더(Paradigm, Beckman, USA)를 이용하여 480 nm에서의 흡광도를 측정하였다. 세포 생존율은 비히클(vehicle, dimethylsulfoxide; DMSO)을 처리한 대조군에 대한 %로 나타내었다.The cytotoxicity of the Baekdugu and Kurage extract prepared in Example 1 was confirmed. 3T3-L1 (American Type Culture Collection; ATCC, USA) was inoculated into DMEM (Dulbecco's Modified Eagle) containing 10% bovine calf serum (BCS) and penicillin / streptomycin Medium) medium. 10 5 cells per well were plated on 24-well tissue culture plates for 24 h and incubated at various concentrations of white rot (50, 100, 150, 200, 300, 400, 500 μg / ml) , 200, 250, 300, and 500 μg / ml), respectively. After 72 hours, the medium containing the sample was removed, replaced with fresh medium containing water-soluble tetrazolium salt (WST) reagent, incubated at 37 ° C for 30 minutes, and incubated for 4 hours using a multiplate reader (Paradigm, Beckman, nm absorbance was measured. Cell viability was expressed as% of vehicle (dimethylsulfoxide; DMSO) treated control.
상기 백두구 또는 희렴 메탄올 추출물을 처리하여 배양한 3T3-L1 지방전구세포의 백두구 및 희렴 추출물 용액 농도에 따른 생존율을 각각 도 1 및 도 2에 나타내었다. 그 결과, 백두구 추출물은 500 μg/ml를 초과하는 농도까지 세포독성을 전혀 유발하지 않는 안전한 물질임을 확인하였으며, 희렴 추출물 역시 300 μg/ml 농도까지 세포독성을 전혀 유발하지 않으며, 500 μg/ml의 농도에서도 80% 이상의 높은 세포 생존율을 나타내는 낮은 독성의 안전한 물질임을 확인하였다.
Survival ratios of 3T3-L1 adipose precursor cells cultured and treated with the Baekdugu or piled methanol extracts according to the concentrations of Baekdugu and Combi extract solutions are shown in Fig. 1 and Fig. 2, respectively. As a result, Baekdugu extract was found to be a safe substance that does not cause any cytotoxicity up to a concentration exceeding 500 μg / ml, and the extract of comvitae did not induce any cytotoxicity up to a concentration of 300 μg / ml. And it was confirmed that it is a low toxic and safe substance exhibiting high cell survival rate of 80% or more even at the concentration.
실시예Example
4: 백두구 및 4: Baekduk and
희렴Celebration
추출물의 지방생성 Fat Production of Extracts
억제능Inhibition
분석 analysis
백두구 및 희렴 추출물의 지방생성 억제효과를 확인하기 위하여, MDI를 이용하여 지방세포로의 분화를 유도하고 그에 따른 지방생성 및 그 억제효과를 확인하였다.In order to confirm the effect of inhibiting the lipogenesis of Baekdugu and Jangmae extracts, MDI was used to induce the differentiation into adipocytes and the lipogenesis and its inhibitory effect were confirmed.
먼저, 3T3-L1 지방전구세포(preadipocyte)를 MDI 즉, 0.5 μM 3-이소부틸-1-메틸크산틴(3-isobuty1-methylxanthine; IBMX), 1 mM 덱사메타손(dexamethasone; DEX) 및 10 μg/ml 인슐린(insulin)을 포함하는 지방형성 분화배지에 배양하여 지방세포(adipocyte)로의 분화를 유도하였다. 12 웰 조직 배양 플레이트에 웰 당 105 개의 세포를 분주하고 2일 후 10% 소태아혈청(fetal bovine serum; FBS)을 포함하는 배지로 교체하였다. 2일 경과 후 IBMX, DEX 및 인슐린을 포함하는 배지로 교환하면서 각각 백두구 또는 희렴 추출물을 농도별로 처리하고 2일 간격으로 총 4회 인슐린 및 상기 추출물을 처리하였다. 마지막 시료를 처리하고 2일 후 지방생성 억제능을 관찰하였다.First, 3T3-L1 preadipocytes were incubated with MDI, 0.5 μM 3-isobuty1-methylxanthine (IBMX), 1 mM dexamethasone (DEX) and 10 μg / ml The cells were cultured in adipogenic differentiation medium containing insulin to induce differentiation into adipocytes. 10 5 cells per well were plated on 12 well tissue culture plates and replaced with medium containing 10% fetal bovine serum (FBS) 2 days later. After 2 days, each of the extracts was treated with a concentration of Baekduju or Rice extract and the insulin and the extract were treated at intervals of 2 days with exchanging the medium with IBMX, DEX and insulin. The final sample was treated and the inhibitory effect on fat production was observed after 2 days.
상기와 같이 MDI로 분화를 유도한 3T3-L1의 지방생성에 대한 효과를 확인하기 위하여, 오일 레드 오 염색을 이용하여 중성지방(triglyceride; TG) 생성을 측정하였다. 지방세포로의 분화 유도 및 시료 처리가 완료된 세포를 1×인산완충염용액(phosphate buffered saline; PBS)으로 세척한 다음 10% 포르말린(formalin)으로 고정하고 오일 레드 오 염색 용액으로 처리한 후 30분간 인큐베이션하였다. 염색이 완료된 후 100% 이소프로판올(isopropanol)을 이용하여 세포에 침착된 염료를 추출하고 520 nm에서 상기 염료 추출액의 흡광도를 측정하였다.In order to confirm the effect of 3T3-L1 induced differentiation by MDI on lipogenesis, triglyceride (TG) production was measured using oil red staining. After induction of differentiation and treatment of adipocytes, the cells were washed with 1 × phosphate buffered saline (PBS), fixed with 10% formalin, treated with oil red staining solution, incubated for 30 minutes Respectively. After the dyeing was completed, the dye deposited on the cells was extracted with 100% isopropanol and the absorbance of the dye extract was measured at 520 nm.
도 3은 (A) 상기 백두구 추출물을 각각 농도별로 처리하여 배양한 3T3-L1의 오일 레드 오 염색결과를 현미경 상으로 확인한 결과 및 (B) 상기 염색된 오일 레드 오 염료를 추출하여 흡광도를 측정하여 계산한 중성지방 함량을 나타낸 결과이다. 도 3에 나타난 바와 같이, 백두구 추출물은 농도 의존적으로 중성지방(triglyceride; TG) 생성을 감소시키며 특히 200 μg/ml의 농도로 처리시 현저히 감소하는 것을 확인하였다. 이러한 결과는 상기 백두구 추출물이 지방생성을 억제하는 효과가 있음을 나타내는 결과이다.FIG. 3 is a graph showing the result of microscopic examination of the result of oil-red staining of 3T3-L1 cultured by treating (A) the Baekdugu extract with concentration, and (B) And the calculated triglyceride content. As shown in FIG. 3, it was confirmed that the Baekdugu extract decreased triglyceride (TG) production in a concentration-dependent manner and markedly decreased especially at a concentration of 200 μg / ml. These results indicate that the Baekdugu extract has an effect of inhibiting lipogenesis.
또한 도 4는 희렴 추출물을 각각 농도별로 처리하여 배양한 3T3-L1의 오일 레드 오 염색결과를 현미경 상으로 확인한 결과 및 상기 염색된 오일 레드 오 염료를 추출하여 흡광도를 측정하여 계산한 중성지방 함량을 나타낸 결과로, 상기 백두구 추출물과 마찬가지로 희렴 추출물이 농도 의존적으로 중성지방 생성을 감소시키는 것을 확인하였다. 이는 상기 희렴 추출물이 지방생성을 억제하는 효과가 있음을 나타내는 것이다.
FIG. 4 is a graph showing the result of microscopic examination of the result of oil-red stain of 3T3-L1 cultured by treatment with each concentration of each extract, and the neutral fat content calculated by measuring the absorbance of the stained oil- As a result, it was confirmed that, like the Baekdugu extract, the tryptic extract reduced the triglyceride production in a concentration-dependent manner. This indicates that the above-described comed extract has an effect of inhibiting lipogenesis.
실시예Example
5: 5:
백두구Baekdugu
및 And
희렴Celebration
추출물의 지방형성 관련 유전자 발현에 대한 효과 Effect of extract on lipogenesis-related gene expression
상기 백두구 및 희렴 추출물의 항비만 활성에 대한 구체적인 기전을 확인하기 위하여 지방형성에 관여하는 핵심인자인 C/EBPα, C/EBPβ 및 PPARγ의 유전자 발현을 분석하였다. 즉, 상기 실시예 4에서 육안으로 관찰한 백두구 및 희렴 추출물의 지방형성 억제를 확인한 것과 더불어 백두구 및 희렴 추출물이 지방형성 관련 유전자의 발현에 미치는 영향을 확인하였다. 이를 위하여, 상기 실시예 4와 동일한 방법으로 세포를 배양하고 각각 백두구 또는 희렴 추출물을 처리한 후, 세포를 회수하여 유전자 발현 분석을 위한 RNA를 분리하여 RT-PCR(reverse-transcriptase polymerase chain reaction)로 분석하였다.In order to confirm the specific mechanism of anti-obesity activity of Baekdugu and Coma extracts, gene expression of C / EBPα, C / EBPβ and PPARγ, which are key factors involved in lipogenesis, was analyzed. Namely, the inhibition of lipid formation of Baekduju and Rice extract was visually observed in Example 4, and the effect of Baekdugu and Rice extract on the expression of lipogenesis-related genes was confirmed. For this, the cells were cultured in the same manner as in Example 4, treated with Baekduju or Rice extract, respectively, and the cells were recovered to isolate RNA for gene expression analysis and subjected to reverse-transcriptase polymerase chain reaction (RT-PCR) Respectively.
먼저, RT-PCR을 위해 백두구 또는 희렴 추출물을 처리한 배양 세포에서 RNA를 추출하여 NanoVue plus 분광광도계(GE Healthcare)로 흡광도를 측정하여 추출된 RNA 농도를 정량한 후 SuperScriptTM First-Strand Synthesis System(Invitrogen, CA, USA)을 이용하여 cDNA를 합성하여 PCR을 수행하였다.First, RNA was extracted from cultured cells treated with Baekduduk or Jangmae extract for RT-PCR, and the absorbance was measured with a NanoVue plus spectrophotometer (GE Healthcare) to determine the concentration of extracted RNA. Then, SuperScript ™ First-Strand Synthesis System Invitrogen, CA, USA) to perform PCR.
도 5는 농도별로 백두구 추출물을 처리하여 배양한 세포에서 지방형성 관련 유전자 발현 정도를 확인한 결과이다. 도 5에 나타낸 바와 같이, MDI 처리에 의해 지방분화가 일어나는 대조군에서는 세 가지 지방형성 관여 유전자(C/EBPα, C/EBPβ 및 PPARγ)의 유의적 증가가 발견된 반면, 백두구 추출물 처리에 의해서는 상기 증가된 유전자들의 발현이 농도 의존적으로 감소하는 것을 확인하였다. 따라서, 본 발명의 백두구 추출물은 지방형성에 관여하는 핵심 인자의 발현을 저해함으로써 지방전구세포가 지방세포로 분화되는 것을 억제함을 확인하였으며, 궁극적으로 항비만 활성을 나타냄을 확인하였다.FIG. 5 shows the results of confirming the degree of expression of lipogenesis-related genes in cells cultured by treatment with Baekdugu extract. As shown in FIG. 5, in the control group in which lipidation was caused by MDI treatment, significant increase of three fat-forming genes (C / EBP ?, C / EBP? And PPAR?) Was found, And the expression of the genes was decreased in a concentration-dependent manner. Therefore, it has been confirmed that the extract of Baekduguchi of the present invention inhibits the expression of the key factor involved in lipogenesis, thereby inhibiting the differentiation of adipocyte precursor cells into adipocytes, and ultimately shows anti-obesity activity.
또한, 도 6은 농도별로 희렴 추출물을 처리하여 배양한 세포에서 지방형성 관련 유전자 발현 정도를 확인한 결과이다. 상기 백두구 추출물에 대한 결과와 유사하게 지방형성 관여 유전자의 발현 증가도가 처리한 희렴 추출물의 농도에 의존적으로 감소하는 것을 확인하였다. 따라서, 희렴 추출물 역시 지방형성에 관여하는 핵심 인자의 발현을 저해함으로써 지방전구세포가 지방세포로 분화되는 것을 억제함을 확인하였으며, 궁극적으로 항비만 활성을 나타냄을 확인하였다.
FIG. 6 shows the results of confirming the degree of expression of lipogenesis-related genes in cells cultured by treatment with a recombinant extract at different concentrations. Similar to the results of the Baekduguchi extract, the expression level of the lipogenic gene was decreased depending on the concentration of the extract. Therefore, it was confirmed that the comedium extract also inhibited the expression of the key factor involved in lipogenesis, thereby inhibiting the differentiation of lipid precursor cells into adipocytes, and ultimately showed anti-obesity activity.
실시예Example
6: 백두구 및 6: Baekdugu and
희렴Celebration
추출 혼합물의 시너지적 Synergistic of the extraction mixture
항비만Anti-obesity
효과 effect
각각에 대해서 항비만 활성이 확인된 백두구 및 희렴 추출물을 1:1로 혼합하여 상기 실시예 4와 동일한 방법으로 항비만 활성을 확인하고, 그 결과를 도 7에 나타내었다. 도 7에 나타난 바와 같이, 지방생성 억제능은 각각의 추출물을 1:1로 혼합한 혼합물을 200 μg/ml로 사용하였을 때 현저한 시너지적 효과를 나타냄을 확인하였다.
The results are shown in FIG. 7. The results are shown in FIG. 7, wherein the anti-obesity activity was confirmed in the same manner as in Example 4, except that Baekduju and Comvitae extracts were mixed at a ratio of 1: 1. As shown in FIG. 7, it was confirmed that the effect of inhibiting lipogenesis was remarkably synergistic when 200 μg / ml of the mixture of each extract was mixed at a ratio of 1: 1.
Claims (8)
A pharmaceutical composition for the prevention or treatment of obesity, comprising a Baekdugu extract.
상기 추출물은 C1 내지 C4의 저급 알콜 또는 이들의 혼합용매로 추출한 것인 조성물.
The method according to claim 1,
Wherein the extract is extracted with a lower alcohol of C1 to C4 or a mixed solvent thereof.
상기 추출물은 메탄올을 용매로 사용하여 65 내지 85℃의 온도에서 제조되는 것인 조성물.
The method according to claim 1,
Wherein the extract is prepared at a temperature of 65-85 < 0 > C using methanol as a solvent.
상기 비만 예방 또는 치료는 지방전구세포의 지방세포로의 분화 억제에 의해 달성되는 것인 조성물.
The method according to claim 1,
Wherein the prevention or treatment of obesity is achieved by inhibiting differentiation of adipose precursor cells into adipocytes.
상기 조성물은 체중감소, 체지방감소, 혈중 중성지방 함량 감소 및 혈중 콜레스테롤 함량 감소의 효과를 가지는 것이 특징인 조성물.
The method according to claim 1,
Wherein said composition has the effect of reducing weight, reducing body fat, reducing blood triglyceride content, and reducing blood cholesterol content.
희렴 추출물을 추가로 포함하는 것인 조성물.
The method according to claim 1,
Lt; RTI ID = 0.0 > pleiotropic < / RTI > extract.
A health functional food for prevention or improvement of obesity comprising Baekdugu extract as an active ingredient.
희렴 추출물을 추가로 포함하는 것인 건강기능식품.
8. The method of claim 7,
A health functional food which further comprises a flavor extract.
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| KR102541902B1 (en) | 2023-03-29 | 2023-06-13 | 엘앤피코스메틱 주식회사 | Method for producing cardamom seed oil and a cosmetic composition for skin cooling and anti-inflammation containing cardamom seed oil obtained thereof |
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| JP2005126405A (en) | 2003-10-25 | 2005-05-19 | Sadami Ishibashi | Antiobesity drug |
| KR100706283B1 (en) | 2005-05-23 | 2007-04-11 | 학교법인 인제학원 | Pharmaceutical composition comprising the extract of Amomum xanthiodes Wallich for treating or preventing obesity and improving lipid metabolism |
| JP2009249331A (en) | 2008-04-04 | 2009-10-29 | Kaneka Corp | Plant-originated agent for preventing or ameliorating hyperlipemia |
| KR101032313B1 (en) * | 2009-04-01 | 2011-05-06 | 연세대학교 산학협력단 | Composition for Preventing or Treating of Obesity, Dyslipidemia, Fatty Liver or Insulin Resistance Syndrome Comprising Camphor as Active Ingredients |
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