KR102070970B1 - Composition comprising extraction of Acer palmatum for preventing or treating enteropathy - Google Patents
Composition comprising extraction of Acer palmatum for preventing or treating enteropathy Download PDFInfo
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- KR102070970B1 KR102070970B1 KR1020180057413A KR20180057413A KR102070970B1 KR 102070970 B1 KR102070970 B1 KR 102070970B1 KR 1020180057413 A KR1020180057413 A KR 1020180057413A KR 20180057413 A KR20180057413 A KR 20180057413A KR 102070970 B1 KR102070970 B1 KR 102070970B1
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- intestinal
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- maple
- mice
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Abstract
본 발명은 단풍나무 추출물을 포함하는 장 질환의 예방 또는 치료용 약학 조성물, 단풍나무 추출물을 인간을 제외하는 개체에 투여하는 단계를 포함하는 장 질환의 예방 또는 치료방법 및 단풍나무 추출물을 포함하는 장 질환의 예방 또는 개선용 건강기능식품에 관한 것이다. 본 발명에 따른 단풍나무 추출물은 장내 밀착 연접을 구성하는 ZO-1 및 오클루딘 단백질의 합성을 촉진시킴으로 장내 투과성을 증가시키고, 장 질환에 의한 체중 감소 및 대장 길이 감소를 저해시키는 효과를 보이므로, 장 질환 예방 또는 치료용 약학 조성물 또는, 예방 또는 개선용 건강기능식품으로 유용하게 사용될 수 있다.The present invention provides a pharmaceutical composition for preventing or treating intestinal diseases, including maple extract, a method for preventing or treating intestinal diseases, comprising administering maple extract to a subject excluding humans, and a bowel containing maple extract. It relates to a dietary supplement for the prevention or improvement of diseases. Maple extract according to the present invention increases the intestinal permeability by promoting the synthesis of the ZO-1 and ocludine protein constituting the intestinal tight junction, and because it has the effect of inhibiting weight loss and colon length reduction due to intestinal diseases, It may be usefully used as a pharmaceutical composition for preventing or treating intestinal diseases or as a nutraceutical for preventing or improving.
Description
본 발명은 단풍나무 추출물을 포함하는 장 질환의 예방 또는 치료용 약학 조성물, 단풍나무 추출물을 인간을 제외하는 개체에 투여하는 단계를 포함하는 장 질환의 예방 또는 치료방법 및 단풍나무 추출물을 포함하는 장 질환의 예방 또는 개선용 건강기능식품에 관한 것이다.The present invention provides a pharmaceutical composition for preventing or treating intestinal diseases, including maple extract, a method for preventing or treating intestinal diseases, comprising administering maple extract to a subject excluding humans, and a bowel containing maple extract. It relates to a dietary supplement for the prevention or improvement of diseases.
장 질환은 복통을 동반하며 소화 장애를 유발하고 다양한 원인에 의해 발생하는 현대인에게서 흔히 나타나는 질환으로, 대표적으로 과민성 장 증후군과 염증성 장 증후군으로 나뉜다.Intestinal diseases are common in modern people with abdominal pain, causing digestive disorders and being caused by various causes, and are classified into irritable bowel syndrome and inflammatory bowel syndrome.
과민성 장 증후군 (IBS)은 기질적 원인이 없이 장기간 반복되는 복부팽만감 등의 복부 불편감 및 복통과 더불어, 설사, 변비 등의 배변 습관의 변화를 동반하는 만성 질환이며, 그 증상이 정신적인 요인이나 스트레스에 의해서 악화하기도 한다. 증상에 따라 설사 우세형, 변비 우세형, 및 통증 우세형으로 구분하여 약물 치료가 되고 있으며, 한국 국내 환자 중 설사형이 30.8%, 변비형이 24.6%, 설사와 변비 교대형이 44.6%를 차지한다. Irritable bowel syndrome (IBS) is a chronic disease accompanied by changes in bowel habits such as diarrhea and constipation, as well as abdominal discomfort and abdominal pain, such as bloating, which is repeated over a long period of time without any organic causes. May be exacerbated by stress. According to the symptoms, diarrhea predominantly, constipation predominantly, and pain predominantly divided into drug treatments. Among Korean patients, diarrhea 30.8%, constipation 24.6%, diarrhea and constipation shift 44.6%. do.
과민성 장 증후군의 치료를 위한 약물요법은 과민성 장 증후군의 전반적인 증상을 개선하는 약제와 단일 증상에 대한 약제로 나눌 수 있다. 복통에 관한 약물로는 평활근 이완제, 항우울제, 오피오이드 작용제(Opioid agonist) 등이 사용되고, 변비 우세형 과민성 장 증후군에 관한 약제로는 섬유제제, 완화제, 5-HT4 작용제 등이 사용되고 있으며, 설사 우세형 과민성 장 증후군에 관한 약제로는 지사제, 5-HT3 길항제 등이 사용된다. 그러나 기존 5-HT 계열 약물들은 부작용으로 인해 그 사용이 제한되고 있기 때문에 현재까지 과민성 장 증후군에 대한 약물요법은 매우 제한적이고, 따라서 대증요법에 대한 의존도가 높아 만족스럽지 못한 실정이다Drug therapy for the treatment of irritable bowel syndrome can be divided into drugs for improving the general symptoms of irritable bowel syndrome and drugs for single symptoms. Drugs related to abdominal pain include smooth muscle relaxants, antidepressants, opioid agonists, and drugs related to constipation predominant type hypersensitivity intestinal tract syndrome are used such as fiber, mitigating agents, and 5-HT4 agonists. As drugs related to bowel syndrome, anti-diabetic agents and 5-HT3 antagonists are used. However, since the use of the existing 5-HT-based drugs is limited due to side effects, pharmacotherapy for hypersensitivity bowel syndrome is very limited, and therefore, the reliance on symptomatic therapy is unsatisfactory.
한편, 궤양성 대장염이나 크론병은 아직 원인이 불분명한 염증성 장 질환 (IBD)으로서, 치유와 재발을 반복하는 만성 난치성 질환이다. 이들 질환의 병인으로서는 유전적 요인이나 면역 이상뿐만 아니라, 식사 등의 환경인자가 관여한다는 지적이 있지만 원인은 아직 불분명하다. 궤양성 대장염은 대장의 점막에 짓무름이나 궤양이 연속적으로 형성되는 염증성 질환으로, 혈변, 점혈변, 설사, 복통이 일어나고, 중증인 경우에는 발열, 체중감소, 빈혈 등의 전신성의 증상이 나타난다. 또한, 크론병은 입에서 항문에 이르는 소화기관의 어느 부위에나 염증이나 궤양 등의 병변이 비연속적으로 발생하는 것을 특징으로 하며, 복통, 설사, 혈변과 더불어, 중증의 경우에는 발열, 하혈, 체중감소, 전신권태감, 빈혈 등의 증상이 나타난다.Ulcerative colitis or Crohn's disease, on the other hand, is an inflamed bowel disease (IBD) whose cause is still unknown. Although the etiology of these diseases is not only genetic factors and immune abnormalities, but also environmental factors such as eating are pointed out, the cause is still unclear. Ulcerative colitis is an inflammatory disease that causes sores or ulcers to form continuously on the mucosa of the large intestine. Bloody stools, mucosal stools, diarrhea and abdominal pain occur, and in severe cases, systemic symptoms such as fever, weight loss, and anemia appear. In addition, Crohn's disease is characterized by discontinuously occurring lesions such as inflammation or ulceration in any part of the digestive system from the mouth to the anus. In addition to abdominal pain, diarrhea and bloody stool, fever, bleeding and weight Symptoms include decreasedness, general malaise, and anemia.
종래에는 이들 염증성 장 질환의 발병율은 서구권에서 높다고 알려져 있었지만, 최근, 한국에서도 식습관 등의 생활습관의 변화와 함께 환자 수가 급증하고 있다. 그렇지만, 원인이 불분명한 이유도 있어 근본적 치료법은 확립되어 있지 않다. 그 때문에 현재의 치료는, 완치를 목표로 하는 것이 아닌, 증상의 개선을 도모하고, 그것을 장기간 유지하는 것을 목표로 한다. 이러한 치료을 위한 약제로서, 주로 아미노살리실산제제, 부신피질 스테로이드제, 면역억제제 등이 사용되지만, 이들의 부작용은 임상적으로 커다란 문제점을 가지고 있다. 예를 들면, 아미노살리실산제제로서 자주 사용되는 살라조설파피리딘의 부작용에는 구역질, 구토, 식욕부진, 발진, 두통, 간장해, 백혈구 감소, 이상 적혈구, 단백뇨, 설사 등이 보고되고 있다. 또한, 부신피질스테로이드제는 일반적으로는 프레드니솔론의 경구투여, 관장, 좌약, 정맥 주사 등으로 사용되지만, 위궤양이나 장기사용에 의한 대퇴골두 괴사 등의 부작용이 존재한다. 그러나 투약의 중단은 증상의 재발을 불러오기 때문에, 이들 약제는 계속적으로 사용하지 않을 수 없다.Conventionally, the incidence of these inflammatory bowel diseases is known to be high in the western region, but in recent years, the number of patients has increased rapidly with changes in lifestyles such as eating habits. However, there are some reasons why the cause is unclear, and no fundamental treatment is established. Therefore, the present treatment does not aim at complete cure but aims at improving symptoms and maintaining it for a long period of time. As a medicament for such treatment, aminosalicylic acid agents, corticosteroids, immunosuppressants and the like are mainly used, but their side effects have clinical problems. For example, side effects of salazosulpapyridine, which is frequently used as an aminosalicylic acid agent, have been reported for nausea, vomiting, anorexia, rash, headache, hepatic injuries, white blood cell reduction, abnormal red blood cells, proteinuria, diarrhea and the like. In addition, corticosteroids are generally used by oral administration of prednisolone, enema, suppositories, intravenous injections, etc., but there are side effects such as femoral head necrosis due to gastric ulcer and intestinal organ use. However, because discontinuation of medications leads to a relapse of symptoms, these drugs cannot be used continuously.
이에, 본 발명자들은 상기와 같은 장 질환에 효과적인 동시에 부작용이 작은 천연물을 개발하기 위해 예의 연구 노력한 결과, 단풍나무 추출물의 장내 밀착 연접(tight junction)을 구성하는 ZO-1 및 오클루딘(occludin) 단백질의 합성을 촉진하여 장내 투과성을 증가시키고, 염증성 장 질환에 의한 체중 감소 및 대장 길이 감소를 저해시키는 효과를 확인하여 본 발명을 완성하였다.Accordingly, the present inventors have made intensive studies to develop natural products that are effective against the above intestinal diseases and have small side effects. As a result, ZO-1 and occludin proteins constituting a tight junction of maple extracts The present invention was completed by confirming the effect of promoting the synthesis of the intestinal permeability, inhibiting the weight loss and length reduction of the colon due to inflammatory bowel disease.
삭제delete
본 발명의 하나의 목적은 단풍나무 추출물을 포함하는 장 질환 예방 또는 치료용 약학 조성물을 제공하는 것이다.One object of the present invention to provide a pharmaceutical composition for preventing or treating intestinal diseases, including maple extract.
본 발명의 다른 하나의 목적은 단풍나무 추출물을 인간을 제외한 개체에 투여하는 단계를 포함하는 장 질환 예방 또는 치료방법을 제공하는 것이다.Another object of the present invention is to provide a method for preventing or treating intestinal diseases comprising administering maple extract to a subject other than a human.
본 발명의 또 다른 하나의 목적은 단풍나무 추출물을 포함하는 장 질환 예방 또는 개선용 건강기능식품을 제공하는 것이다.Another object of the present invention to provide a dietary supplement for the prevention or improvement of intestinal diseases comprising maple extract.
본 발명의 또 다른 하나의 목적은 단풍나무 추출물을 포함하는 장 질환 예방 또는 개선용 사료 조성물을 제공하는 것이다.Still another object of the present invention is to provide a feed composition for preventing or improving intestinal diseases, including maple extract.
상기의 목적을 달성하기 위한 본 발명의 하나의 양태는, 단풍나무 추출물을 포함하는 장 질환 예방 또는 치료용 약학 조성물을 제공한다.One aspect of the present invention for achieving the above object, provides a pharmaceutical composition for preventing or treating intestinal diseases, including maple extract.
본 발명에서의 용어, "단풍나무(Acer palmatum)"는 무환자나무과(Sapindaceae)의 식물로 산지의 계곡에서 자란다. 높이는 10m에 달하고, 작은 가지는 털이 없으며 붉은빛을 띤 갈색이다. 잎은 마주나고 손바닥 모양으로 5∼7개로 깊게 갈라진다. 갈라진 조각은 넓은 바소 모양이고 끝이 뾰족하며 가장자리에 겹톱니가 있고 길이가 5∼6cm이다. 잎자루는 붉은 색을 띠고 길이가 3∼5cm이다. 단풍나무는 주로 관상용으로 심으며 땔감으로 쓰인다. 한방에서 뿌리 껍질과 가지를 계조축(鷄爪)이라는 약재로 쓰는데, 무릎관절염으로 통증이 심할 때, 물에 넣고 달여서 복용하고, 골절상을 입었을 때 오가피를 배합해서 사용하며, 소염 작용과 해독 효과가 있다. 한국(제주, 전남, 전북) 및 일본에 분포한다.The term " Acer palmatum " in the present invention is a plant of Sapindaceae and grows in the valleys of the mountains. Its height is 10m and small branches are hairless and reddish brown. The leaves face each other and are deeply divided into 5-7 in the shape of palms. The split pieces are wide lanceolate, pointed at the end, with coarsely on the edge, 5-6cm long. Petioles are red in color and are 3-5cm long. Maple trees are mainly planted for ornamental purposes and used as firewood. Root bark and eggplant is used as a medicine called gradation axis (축) in oriental medicine. When pain is severe due to knee arthritis, put it in water and put it in the water. have. It is distributed in Korea (Jeju, Jeonnam, Jeonbuk) and Japan.
본 발명에서의 단풍나무는 추출물 생성을 위해 그의 잎, 가지 또는 뿌리를 모두 사용할 수 있고, 구체적으로는 잎과 가지를 포함한 지상부를 각각 사용할 있으며, 보다 구체적으로는 잎을 사용할 수 있으나 이에 한정되는 것은 아니다. 아울러, 상기 단풍나무는 채취한 것, 재배한 것 또는 시판되는 것 등의 제한이 없이 사용 가능하다. Maple trees in the present invention can use all of its leaves, branches or roots for the production of extracts, specifically used above ground parts including leaves and branches, more specifically can use leaves, but is not limited thereto. no. In addition, the maple can be used without limitation, such as harvested, grown or commercially available.
상기 단풍나무 추출물은 단풍나무 조추출물뿐 아니라 추출물의 추가적인 가공에 의한 모든 형태를 의미한다. The maple extract refers to all forms by additional processing of the extract as well as crude maple extract.
상기 단풍나무 추출물은 단풍나무 그 자체 또는 이의 분쇄물이거나, 추출공정을 통해 수득한 추출물일 수 있다. 상기 추출공정은 당해 기술분야에서 이용되는 통상의 방법에 따라 실시될 수 있다.The maple extract may be maple itself or a pulverized product thereof, or an extract obtained through an extraction process. The extraction process may be carried out according to conventional methods used in the art.
상기 추출 용매는 물, 알코올 또는 이의 혼합물, 구체적으로는 C1 내지 C4의 저급 알코올 또는 이들의 혼합 용매로부터 선택된 용매를 사용하는 것일 수 있으나, 이에 한정되는 것은 아니다. 상기 추출 용매의 양은 단풍나무의 건조 중량의 2 내지 20 배로 하는 것일 수 있으나 이에 한정되지 않는다. The extraction solvent may be a solvent selected from water, an alcohol or a mixture thereof, specifically, a lower alcohol of C 1 to C 4 or a mixed solvent thereof, but is not limited thereto. The amount of the extraction solvent may be 2 to 20 times the dry weight of the maple tree, but is not limited thereto.
상기 추출 방법은 열수추출, 침지 추출, 환류 냉각 추출 및 초음파 추출 등 의 추출 방법을 사용할 수 있으며, 구체적으로는 침지 추출방법으로 추출될 수 있다. 추출시 온도는 구체적으로 10℃ 내지 100℃일 수 있으며, 보다 구체적으로 상온일 수 있으나 이에 한정되지 않는다. 상기 추출 시간은 구체적으로 6시간 내지 7일일 수 있고, 보다 구체적으로 12시간 내지 3일일 수 있으나 이에 한정되지 않는다.The extraction method may be an extraction method such as hot water extraction, immersion extraction, reflux cooling extraction and ultrasonic extraction, specifically, can be extracted by immersion extraction. In the extraction, the temperature may be specifically 10 ° C. to 100 ° C., more specifically, room temperature, but is not limited thereto. The extraction time may be specifically 6 hours to 7 days, more specifically 12 hours to 3 days, but is not limited thereto.
본 명세서에서, 상기 단풍나무 추출물은 KIOM-2015E와 혼용하여 사용될 수 있다.In the present specification, the maple extract may be used in combination with KIOM-2015E.
상기 장 질환은 과민성 장 증후군 또는 염증성 장 질환일 수 있다.The bowel disease may be irritable bowel syndrome or inflammatory bowel disease.
상기 과민성 장 증후군 (IBS)은 기질적 원인이 없이 장기간 반복되는 복부팽만감 등의 복부 불편감 및 복통과 더불어, 설사, 변비 등의 배변 습관의 변화를 동반하는 만성 질환이며, 그 증상이 정신적인 요인이나 스트레스에 의해서 악화하기도 한다. 증상에 따라 설사 우세형, 변비 우세형, 및 통증 우세형으로 구분할 수 있다.Irritable bowel syndrome (IBS) is a chronic disease accompanied by changes in bowel habits such as diarrhea and constipation, along with abdominal discomfort and abdominal pain such as abdominal bloating, which is repeated over a long period of time without an organic cause. It can also be exacerbated by stress or stress. Depending on the symptoms, it can be divided into diarrhea dominant type, constipation dominant type, and pain dominant type.
상기 염증성 장 질환은 장 내에 염증이 발생함으로써 장내 기능에 장애가 생기는 질환으로, 치유와 재발을 반복하는 만성 난치성 질환이다. 이들 질환의 병인으로서는 유전적 요인이나 면역 이상뿐만 아니라, 식사 등의 환경인자가 관여한다는 지적이 있지만 원인은 아직 불분명하다. 염증성 장 질환은 대표적으로 궤양성 대장염 및 크론병이 있으며, 궤양성 대장염은 대장의 점막에 짓무름이나 궤양이 연속적으로 형성되는 염증성 질환으로, 혈변, 점혈변, 설사, 복통이 일어나고, 중증인 경우에는 발열, 체중감소, 빈혈 등의 전신성의 증상이 나타난다. 또한, 크론병은 입에서 항문에 이르는 소화기관의 어느 부위에나 염증이나 궤양 등의 병변이 비연속적으로 발생하는 것을 특징으로 하며, 복통, 설사, 혈변과 더불어, 중증의 경우에는 발열, 하혈, 체중감소, 전신권태감, 빈혈 등의 증상이 나타난다.The inflammatory bowel disease is a disease in which intestinal function is impaired by inflammation occurring in the intestine, and is a chronic refractory disease that repeats healing and recurrence. Although the etiology of these diseases is not only genetic factors and immune abnormalities, but also environmental factors such as eating are pointed out, the cause is still unclear. Inflammatory bowel disease is typical of ulcerative colitis and Crohn's disease. Ulcerative colitis is an inflammatory disease that causes sores or ulcers to form continuously in the mucous membrane of the colon. Bloody stools, mucosal stools, diarrhea and abdominal pain occur in severe cases. Systemic symptoms such as fever, weight loss, and anemia appear. In addition, Crohn's disease is characterized by discontinuously occurring lesions such as inflammation or ulceration in any part of the digestive system from the mouth to the anus. In addition to abdominal pain, diarrhea and bloody stool, fever, bleeding and weight Symptoms include decreasedness, general malaise, and anemia.
본 발명에 따른 조성물은 체중 감소를 억제하여 장 질환 치료를 달성하는 것일 수 있다.The composition according to the present invention may be one that inhibits weight loss to achieve intestinal disease treatment.
본 발명의 구체적인 일 실시예에서는, DSS로 염증성 장 질환을 유도한 마우스 중 대조군을 제외한 실험군에 단풍나무 추출물을 처리하고 체중의 변화를 측정한 결과, 장 질환 유도 마우스는 정상군 대비 체중의 감소가 나타났으나, 단풍나무 추출물을 투여한 집단은 정상군과 유사한 수준까지 체중의 감소가 완화된 것을 확인하였다 (도 1).In a specific embodiment of the present invention, as a result of measuring maple weight in the experimental group except for the control group among the mice induced inflammatory bowel disease with DSS, the bowel disease induced mice had a weight loss compared to the normal group. However, the group administered with the maple extract was confirmed that the weight loss was alleviated to a level similar to that of the normal group (FIG. 1).
본 발명에 따른 조성물은 대장의 길이 감소를 억제하여 장 질환 치료를 달성하는 것일 수 있다.The composition according to the invention may be to inhibit the reduction in the length of the large intestine to achieve treatment of intestinal diseases.
본 발명의 다른 구체적인 일 실시예에서는, 염증성 장 질환을 유도한 마우스 중 대조군을 제외한 실험군에 단풍나무 추출물을 처리하고 대장을 적출하여 조직의 대장의 길이를 측정한 결과, 장 질환 유도 마우스는 정상군 대비 매우 짧은 길이의 대장을 갖는 것을 확인하였고, 단풍나무 추출물 투여군은 장 질환 유도군과 비교하여 대장의 길이 감소가 완화된 것을 확인하였다 (도 2).In another specific embodiment of the present invention, as a result of measuring the length of the tissue by treating the maple extract in the experimental group except the control group and extracting the large intestine among the mice that induced inflammatory bowel disease, the intestinal disease-inducing mouse was a normal group. It was confirmed to have a very short length of the large intestine, and the maple extract administration group was confirmed that the reduction in the length of the colon compared to the intestinal disease induction group (Fig. 2).
본 발명에 따른 조성물은 염증을 감소시켜 장 질환 치료를 달성하는 것일 수 있다.The composition according to the invention may be to reduce inflammation to achieve intestinal disease treatment.
본 발명의 또 다른 구체적인 일 실시예에서는, 염증성 장 질환을 유도한 마우스 중 대조군을 제외한 실험군에 단풍나무 추출물을 처리하고 대장을 적출하여 대장 조직 병변을 확인한 결과, 장 질환 유도 마우스는 정상군 대비 대장 조직의 염증이 많이 발생한 것을 확인한 반면, 단풍나무 추출물 투여군은 장 질환 유도군과 비교하여 조직의 염증이 감소된 것을 확인하였다 (도 3).In another specific embodiment of the present invention, as a result of treating maple tissue in the experimental group except the control group of the mice that induced inflammatory bowel disease and extracting the colon, and confirmed colon tissue lesions, the bowel disease-induced mice compared to the normal group. While it was confirmed that a lot of inflammation of the tissue, the maple extract administration group was confirmed that the inflammation of the tissue is reduced compared to the intestinal disease induction group (Fig. 3).
본 발명에 따른 조성물은 대식세포의 침투 억제를 통해 장 질환 치료를 달성하는 것일 수 있다.The composition according to the present invention may be to achieve the treatment of intestinal diseases through the inhibition of infiltration of macrophages.
본 발명의 또 다른 구체적인 일 실시예에서는, 염증성 장 질환을 유도한 마우스 중 대조군을 제외한 실험군에 단풍나무 추출물을 처리하고 루시퍼레이즈를 포함하는 대식세포를 혈관주입으로 마우스에 침투시켜 대장 조직으로의 침투를 확인한 결과, 장 질환 유도군은 정상군과 비교하여 대장 조직의 대식세포 침투가 증가한 것을 확인하였으나, 단풍나무 추출물 투여군은 장 질환 유도군과 비교하여 대식세포의 침투가 감소한 것을 확인하였다 (도 4).In another specific embodiment of the present invention, the inflammatory bowel disease-induced infiltration into the colon tissue by treating the maple extract to the experimental group except the control group and injecting macrophages containing luciferase into the mouse by vascular injection. As a result, the intestinal disease induction group was confirmed that the macrophage infiltration of the colon tissue increased compared to the normal group, the maple extract administration group was confirmed that the infiltration of macrophages is reduced compared to the intestinal disease induction group (Fig. 4 ).
본 발명에 따른 조성물은 염증성 사이토카인의 분비 억제를 통해 장 질환 치료를 달성하는 것일 수 있다.The composition according to the present invention may be one that achieves intestinal disease treatment through inhibition of the secretion of inflammatory cytokines.
본 발명의 또 다른 구체적인 일 실시예에서는, 염증성 장 질환을 유도한 마우스의 혈액을 채취하여 염증성 사이토카인의 분비량을 측정한 결과, 장 질환 유도군에서는 정상군과 비교하여 사이토카인의 분비량이 증가하였으나, 단풍나무 추출물 투여군은 장 질환 유도군과 비교하여 사이토카인의 분비량이 감소한 것을 확인하였다 (도 5).In another specific embodiment of the present invention, as a result of measuring the secretion amount of inflammatory cytokines by collecting blood from mice induced with inflammatory bowel disease, the amount of cytokine secretion was increased in the intestinal disease inducing group compared with the normal group. , Maple extract administration group was confirmed that the amount of cytokine secretion is reduced compared to the intestinal disease induction group (FIG. 5).
본 발명에 따른 조성물은 장내 밀착 연접을 보호하여 장 질환 치료를 달성하는 것일 수 있다.The composition according to the invention may be to protect the intestinal tight junctions to achieve intestinal disease treatment.
본 발명의 또 다른 구체적인 일 실시예에서는, 장 질환을 유도한 마우스의 대장조직에서 밀착 연접과 관련된 단백질의 발현량을 측정한 결과, 장 질환 유도군에서는 정상군과 비교하여 상기 단백질의 발현량이 감소하였으나, 단풍나무 추출물 투여군은 장 질환 유도군과 비교하여 상기 단백질의 발현량이 증가한 것을 확인하였다 (도 6).In another specific embodiment of the present invention, as a result of measuring the expression level of the protein associated with tight junctions in the colon tissue of the mouse induced intestinal disease, the expression level of the protein is reduced in the intestinal disease induction group compared to the normal group However, maple extract administration group was confirmed that the expression level of the protein increased compared to the intestinal disease induction group (Fig. 6).
본 발명에서의 용어 "예방"이란, 본 발명에 따른 단풍나무 추출물을 포함하는 조성물의 투여로 장 질환의 증상을 억제 또는 지연시키는 모든 행위를 말한다.The term "prevention" in the present invention refers to any action that inhibits or delays the symptoms of intestinal disease by administration of a composition comprising the maple extract according to the present invention.
본 발명에서의 용어 "치료"란, 본 발명에 따른 단풍나무 추출물을 포함하는 조성물의 투여로 상기 장 질환의 증상이 호전되거나 이롭게 변경되는 모든 행위를 말한다.The term "treatment" in the present invention refers to any action in which the symptoms of the bowel disease are improved or beneficially changed by administration of the composition containing the maple extract according to the present invention.
본 발명에서 사용된 용어 "약학 조성물"이란, 질병의 예방 또는 치료를 목적으로 제조된 것을 의미하며, 각각의 통상의 방법에 따라 다양한 형태로 제형화하여 사용될 수 있다. 예컨대, 산제, 과립제, 정제, 캡슐제, 현탁액, 에멀젼, 시럽 등의 제형으로 제형화할 수 있고, 외용제, 좌제 및 멸균 주사용액의 형태로 제형화하여 사용될 수 있다. 구체적으로, 점안투여하기 적합한 형태, 예를 들어, 점안제, 크림제, 연고제, 겔제 또는 로션제로 제형화하여 사용될 수 있다.As used herein, the term "pharmaceutical composition" means that is prepared for the purpose of preventing or treating a disease, and can be used by formulating in various forms according to each conventional method. For example, it may be formulated in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, and the like, and may be used in the form of external preparations, suppositories, and sterile injectable solutions. In particular, it may be used in a form suitable for topical administration, for example, eye drops, creams, ointments, gels or lotions.
상기 본 발명의 조성물은, 약학적 조성물의 제조에 통상적으로 사용하는 적절한 담체, 부형제 또는 희석제를 추가로 포함하는 창상 예방 또는 치료용 약학 조성물의 형태로 제조될 수 있는데, 상기 담체는 비자연적 담체(non-naturally occuring carrier)를 포함할 수 있다. The composition of the present invention may be prepared in the form of a pharmaceutical composition for preventing or treating wounds further comprising a suitable carrier, excipient or diluent commonly used in the manufacture of the pharmaceutical composition, wherein the carrier is an unnatural carrier ( non-naturally occuring carriers).
본 발명에서, 상기 약학 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.In the present invention, carriers, excipients and diluents that can be included in the pharmaceutical composition include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, Calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil.
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 상엽 추출물과 이의 분획물들에 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, 올리브 오일과 같은 식물성 기름, 에틸올레이트와 같은 주사 가능한 에스테르 등이 사용될수 있다. 좌제의 기제로는 위텝솔(witepsol), 마크로골, 트윈(tween) 61, 카카오지, 라우린지, 글리세로제라틴 등이 사용될 수 있다.When formulated, diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents and surfactants are usually used. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and the solid preparations include at least one excipient such as starch, calcium carbonate, and the like in the above extracts and fractions thereof. , Sucrose or lactose, gelatin and the like are mixed and prepared. In addition to simple excipients, lubricants such as magnesium styrate and talc are also used. Liquid preparations for oral use may include various excipients such as wetting agents, sweeteners, fragrances, preservatives, etc., in addition to water and liquid paraffin, which are commonly used to include suspensions, solvents, emulsions, and syrups. have. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, suppositories. As the non-aqueous solvent and suspending agent, propylene glycol, polyethylene glycol, vegetable oils such as olive oil, injectable esters such as ethyl oleate and the like can be used. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerogelatin and the like can be used.
본 발명의 약학 조성물은 목적하는 방법에 따라 경구, 비경구, 직장, 국소, 경피, 정맥 내, 근육 내, 복강 내, 피하 등으로 투여할 수 있으며, 약학 조성물의 유효성분은 투여받을 대상의 연령, 성별, 체중, 병리 상태 및 그 심각도, 투여 경로 또는 처방자의 판단에 따라 달라질 것이다. 이러한 인자에 기초한 적용량 결정은 당업자의 수준 내에 있으며, 이의 1일 투여 용량은 예를 들어 0.1 mg/g/일 내지 100 mg/g/일, 보다 구체적으로는 5 mg/g/일 내지 50 mg/g/일이 될 수 있고, 본 발명의 조성물의 투여빈도는 특별히 이에 제한되지 않으나, 1일 1회 투여하거나 또는 용량을 분할하여 수회 투여할 수 있다. 상기 투여량은 어떠한 면으로든 본 발명의 범위를 한정하는 것은 아니다.The pharmaceutical composition of the present invention may be administered orally, parenterally, rectally, topically, transdermally, intravenously, intramuscularly, intraperitoneally, subcutaneously, etc. according to the desired method, and the active ingredient of the pharmaceutical composition may be administered to the age of the subject to be administered. , Sex, weight, pathology and severity, route of administration or prescriber's judgment. Dosage determination based on these factors is within the level of one of skill in the art and its daily dosage may be, for example, from 0.1 mg / g / day to 100 mg / g / day, more specifically from 5 mg / g / day to 50 mg / day. g / day, and the frequency of administration of the composition of the present invention is not particularly limited, but may be administered once a day or several times in divided doses. The dosage does not limit the scope of the invention in any aspect.
본 발명의 다른 하나의 양태는, 단풍나무 추출물을 인간을 제외한 개체에 투여하는 단계를 포함하는 장 질환의 예방 또는 치료방법을 제공한다.Another embodiment of the present invention provides a method for preventing or treating intestinal diseases, comprising administering maple extract to a subject other than a human.
본 발명에서의 용어, "개체"란, 장 질환이 발병하었거나 발병할 가능성이 있는 인간을 포함한 모든 동물을 의미할 수 있다. 상기 동물은 인간뿐만 아니라 이와 유사한 증상의 치료를 필요로 하는 소, 말, 양, 돼지, 염소, 낙타, 영양, 개, 고양이 등의 포유동물일 수 있으나, 이에 제한되지는 않는다.As used herein, the term "individual" may mean any animal, including a human, who has or is likely to develop intestinal disease. The animal may be a mammal such as, but not limited to, a human, a cow, a horse, a sheep, a pig, a goat, a camel, a antelope, a dog, a cat, and the like, which require treatment of similar symptoms.
본 발명의 상기 예방 또는 치료방법은 구체적으로, 장 질환이 발병하였거나 발병할 위험이 있는 개체에 상기 조성물을 약학적으로 유효한 양으로 투여하는 단계를 포함할 수 있다.The method of preventing or treating the present invention may specifically include administering the composition in a pharmaceutically effective amount to a subject having or at risk of developing intestinal disease.
본 발명에서의 용어, "투여"는 어떠한 적절한 방법으로 환자에게 본 발명의 약학 조성물을 도입하는 것을 의미하며, 본 발명의 조성물의 투여 경로는 목적 조직에 도달할 수 있는 한 다양한 경로를 통하여 투여될 수 있으며, 목적하는 바에 따라 점안 투여, 복강내 투여, 정맥내 투여, 근육내 투여, 피하 투여, 피내 투여, 경피패치투여, 경구 투여, 비내 투여, 폐내 투여, 직장내 투여 등의 경로를 통해 투여 될 수 있고, 구체적으로 경구 투여의 경로를 통해 투여될 수 있다.The term "administration" in the present invention means introducing the pharmaceutical composition of the present invention to a patient in any suitable manner, and the route of administration of the composition of the present invention may be administered through various routes as long as it can reach the desired tissue. If desired, administration may be by administration of instillation, intraperitoneal administration, intravenous administration, intramuscular administration, subcutaneous administration, intradermal administration, transdermal patch administration, oral administration, intranasal administration, pulmonary administration, rectal administration, etc. And may specifically be administered via a route of oral administration.
본 발명의 또 다른 하나의 양태는, 단풍나무 추출물을 포함하는 장 질환 예방 또는 개선용 건강기능식품을 제공한다.Another aspect of the present invention provides a dietary supplement for preventing or improving intestinal diseases, including maple extract.
본 발명에서의 용어, "개선"은 상기 조성물을 경구투여하여 골다공증이 호전되거나 이롭게 변경되는 모든 행위를 의미한다.As used herein, the term "improvement" refers to any action in which osteoporosis is improved or beneficially modified by oral administration of the composition.
건강기능식품(functional food)이란, 특정보건용 식품(food for special health use, FoSHU)과 동일한 용어로, 영양 공급 외에도 생체조절기능이 효율적으로 나타나도록 가공된 의학, 의료효과가 높은 식품을 의미하는데, 상기 식품은 암 전이 억제에 유용한 효과를 얻기 위하여 정제, 캡슐, 분말, 과립, 액상, 환 등의 다양한 형태로 제조될 수 있다.Functional food is the same term as food for special health use (FOSHU), and means foods with high medical and medical effects that have been processed to efficiently display bioregulatory functions in addition to nutrition. In addition, the food may be prepared in various forms such as tablets, capsules, powders, granules, liquids, pills, etc. in order to obtain useful effects in inhibiting cancer metastasis.
본 발명의 건강기능식품은 식품학적으로 허용가능한 담체를 추가로 포함하는 것일 수 있다.The dietary supplement of the present invention may further comprise a food acceptable carrier.
본 발명의 단풍나무 추출물을 포함하는 조성물을 첨가할 수 있는 식품의 종류에는 별다른 제한이 없으며, 예를 들어 각종 음료, 껌, 차, 비타민 복합제, 건강보조 식품류 등이 있다. 상기 식품 조성물에는 암 전이 억제 효과에 방해가 되지 않는 다른 성분을 추가할 수 있으며, 그 종류는 특별히 제한되지 않는다. 예를 들어, 통상의 식품과 같이 여러 가지 생약 추출물, 식품학적으로 허용가능한 식품보조첨가제 또는 천연 탄수화물 등을 추가 성분으로서 함유할 수 있다.There is no restriction | limiting in particular in the kind of food which can add the composition containing the maple extract of this invention, For example, there are various drinks, a gum, tea, a vitamin complex, a health supplement, etc. Other ingredients that do not interfere with the cancer metastasis inhibiting effect can be added to the food composition, and the kind thereof is not particularly limited. For example, various herbal extracts, food acceptable additives, natural carbohydrates, and the like may be contained as additional ingredients, such as conventional foods.
상기 식품보조첨가제는 각 제형의 건강기능식품을 제조하는데 첨가되는 것으로서 당업자가 적절히 선택하여 사용할 수 있다. 예를 들어 여러 가지 영양제, 비타민, 광물 (전해질), 합성 풍미제 및 천연 풍미제 등의 풍미제, 착색제 및 충진제, 펙트산 및 그의 염, 알긴산 및 그의 염, 유기산, 보호성 콜로이드 증점제, pH 조절제, 안정화제, 방부제, 글리세린, 알코올, 탄산음료에 사용되는 탄산화제 등이 포함되지만, 상기 예들에 의해 그 종류가 제한되는 것은 아니다.The food supplement additive is added to prepare the health functional food of each formulation can be used by those skilled in the art as appropriate. For example, various nutrients, vitamins, minerals (electrolytes), flavors such as synthetic and natural flavors, colorants and fillers, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusters , Stabilizers, preservatives, glycerin, alcohols, carbonation agents used in the carbonated beverage, and the like, but the kind is not limited by the above examples.
이때, 상기 식품에 포함되는 추출물의 함량은 특별히 이에 제한되지 않으나, 식품 조성물의 총 중량에 대하여 0.01 내지 100 중량%, 보다 바람직하게는 1 내지 80 중량%로 포함될 수 있다. At this time, the content of the extract contained in the food is not particularly limited, but may be included in 0.01 to 100% by weight, more preferably 1 to 80% by weight relative to the total weight of the food composition.
식품이 음료인 경우에는 100㎖를 기준으로 1 내지 30g, 바람직하게는 3 내지 20g의 비율로 포함될 수 있다. 또한, 상기 조성물은 식품 조성물에 통상 사용되어 냄새, 맛, 시각 등을 향상시킬 수 있는 추가 성분을 포함할 수 있다. 예를 들어, 비타민 A, C, D, E, B1, B2, B6, B12, 니아신(niacin), 비오틴(biotin), 폴레이트(folate), 판토텐산(panthotenic acid) 등을 포함할 수 있다. 또한, 아연(Zn), 철(Fe), 칼슘(Ca), 크롬(Cr), 마그네슘(Mg), 망간(Mn), 구리(Cu) 등의 미네랄을 포함할 수 있다. 또한, 라이신, 트립토판, 시스테인, 발린 등의 아미노산을 포함할 수 있다. 또한, 방부제(소르빈산 칼륨, 벤조산나트륨, 살리실산, 데히드로초산나트륨 등), 살균제(표백분과 고도 표백분, 차아염소산나트륨 등), 산화방지제(부틸히드록시아니졸(BHA), 부틸히드록시톨류엔(BHT) 등), 착색제(타르색소 등), 발색제(아질산 나트륨, 아초산 나트륨 등), 표백제(아황산나트륨), 조미료(MSG 글루타민산나트륨 등), 감미료(둘신, 사이클레메이트, 사카린, 나트륨 등), 향료(바닐린, 락톤류 등), 팽창제(명반, D-주석산수소칼륨 등), 강화제, 유화제, 증점제(호료), 피막제, 검기초제, 거품억제제, 용제, 개량제 등의 식품 첨가물(food additives)을 첨가할 수 있다. 상기 첨가물은 식품의 종류에 따라 선별되고 적절한 양으로 사용된다.When food is a beverage, it may be included in a ratio of 1 to 30 g, preferably 3 to 20 g based on 100 ml. In addition, the composition may include additional ingredients that are commonly used in food compositions to improve the smell, taste, time and the like. For example, it may include vitamins A, C, D, E, B1, B2, B6, B12, niacin, biotin, folate, panthotenic acid, and the like. Also, minerals such as zinc (Zn), iron (Fe), calcium (Ca), chromium (Cr), magnesium (Mg), manganese (Mn), and copper (Cu) may be included. It may also contain amino acids such as lysine, tryptophan, cysteine, valine and the like. In addition, preservatives (potassium sorbate, sodium benzoate, salicylic acid, sodium dehydroacetic acid, etc.), fungicides (bleaching powder and highly bleaching powder, sodium hypochlorite, etc.), antioxidants (butylhydroxyanisol (BHA), butylhydroxytoluene ( BHT), etc.), colorants (such as tar pigments), colorants (such as sodium nitrite, sodium nitrite), bleach (sodium sulfite), seasonings (such as MSG glutamate), sweeteners (ducin, cyclate, saccharin, sodium, etc.) , Food additives such as flavors (vanillin, lactones, etc.), swelling agents (alum, aluminium, D-potassium hydrogenate, etc.), reinforcing agents, emulsifiers, thickeners (pigments), coatings, gum herbicides, foam inhibitors, solvents, and improvers. ) Can be added. The additive is selected according to the type of food and used in an appropriate amount.
본 발명의 건강기능식품은 당업계에서 통상적으로 사용되는 방법에 의하여 제조가능하며, 상기 제조 시에는 당업계에서 통상적으로 첨가하는 원료 및 성분을 첨가하여 제조할 수 있다. 또한 일반 약품과는 달리 식품을 원료로 하여 약품의 장기 복용 시 발생할 수 있는 부작용 등이 없는 장점이 있고, 휴대성이 뛰어날 수 있다.The health functional food of the present invention can be prepared by a method commonly used in the art, and the preparation can be prepared by adding raw materials and ingredients commonly added in the art. In addition, unlike the general medicine has the advantage that there is no side effect that can occur when taking a long-term use of the drug as a raw material, and can be excellent in portability.
본 발명의 또 다른 하나의 양태는, 단풍나무 추출물을 포함하는 장 질환 예방 또는 개선용 사료 조성물을 제공한다.Another aspect of the present invention provides a feed composition for preventing or improving intestinal diseases, including maple extract.
상기 사료 조성물은 사료 첨가제를 포함할 수 있다. 본 발명의 사료첨가제는 사료관리법상의 보조사료에 해당한다.The feed composition may comprise a feed additive. The feed additive of the present invention corresponds to a feed supplement in the Feed Control Act.
본 발명에서 용어, "사료"는 동물이 먹고, 섭취하며, 소화시키기 위한 또는 이에 적당한 임의의 천연 또는 인공 규정식, 한끼식 등 또는 상기 한끼식의 성분을 의미할 수 있다.As used herein, the term "feed" may refer to any natural or artificial diet, one meal, or the like, or a component of the one meal, for the animal to eat, ingest, and digest.
상기 사료의 종류는 특별히 제한되지 아니하며, 당해 기술 분야에서 통상적으로 사용되는 사료를 사용할 수 있다. 상기 사료의 비제한적인 예로는, 곡물류, 근과류, 식품 가공 부산물류, 조류, 섬유질류, 제약 부산물류, 유지류, 전분류, 박류 또는 곡물 부산물류 등과 같은 식물성 사료; 단백질류, 무기물류, 유지류, 광물성류, 유지류, 단세포 단백질류, 동물성 플랑크톤류 또는 음식물 등과 같은 동물성 사료를 들 수 있다. 이들은 단독으로 사용되거나 2 종 이상을 혼합하여 사용될 수 있다.The type of the feed is not particularly limited, and may be used a feed commonly used in the art. Non-limiting examples of the feed may include plant feeds such as cereals, fruits, food processing by-products, algae, fibers, pharmaceutical by-products, oils, starches, gourds or grain by-products; And animal feeds such as proteins, minerals, fats and oils, minerals, fats and oils, single-cell proteins, zooplankton or food and the like. These may be used alone or in combination of two or more thereof.
본 발명에 따른 단풍나무 추출물은 장내 밀착 연접을 구성하는 ZO-1 및 오클루딘 단백질의 합성을 촉진시킴으로 장내 투과성을 증가시키고, 염증성 장 질환에 의한 체중 감소 및 대장 길이 감소를 저해시키는 효과를 보이므로, 염증성 장 질환 예방 또는 치료용 약학 조성물 또는, 예방 또는 개선용 건강기능식품으로 유용하게 사용될 수 있다.Maple extract according to the present invention increases the intestinal permeability by promoting the synthesis of ZO-1 and ocludine protein constituting the intestinal tight junction, and because it shows the effect of inhibiting weight loss and colon length decrease by inflammatory bowel disease It can be usefully used as a pharmaceutical composition for preventing or treating inflammatory bowel disease, or as a health functional food for preventing or improving.
도 1은 본 발명 단풍나무 추출물을 5% DSS로 대장염을 유도한 마우스에 경구 투여한 후 체중을 확인한 그래프이다.
도 2는 본 발명 단풍나무 추출물을 5% DSS로 대장염을 유도한 마우스에 경구 투여한 후 대장 길이를 확인한 이미지 및 그래프이다
도 3은 본 발명 단풍나무 추출물을 5% DSS로 대장염을 유도한 마우스에 경구 투여한 후 내시경 및 대장 조직을 염색한 이미지이다.
도 4는 본 발명 단풍나무 추출물을 5% DSS로 대장염을 유도한 마우스에 경구 투여한 후 염증성 대식세포의 침투를 확인한 이미지이다.
도 5는 본 발명 단풍나무 추출물을 5% DSS로 대장염을 유도한 마우스에 경구 투여한 후 염증성 사이토카인의 양을 확인한 그래프이다.
도 6은 본 발명 단풍나무 추출물을 5% DSS로 대장염을 유도한 마우스에 경구 투여한 후 ZO-1의 발현 증가를 보여주는 이미지 및 ZO-1 및 오클루딘의 발현양을 보여주는 웨스턴 블롯 이미지이다. 1 is a graph confirming the weight after oral administration of the present invention maple extract to mice induced colitis with 5% DSS.
Figure 2 is an image and a graph confirming the colon length after oral administration of the present invention maple extract to mice with colitis induced by 5% DSS
Figure 3 is an image staining the endoscope and colon tissue after oral administration of the present invention maple extract to mice induced colitis with 5% DSS.
Figure 4 is an image confirming the infiltration of inflammatory macrophages after oral administration of the present invention maple extract to mice induced colitis with 5% DSS.
Figure 5 is a graph confirming the amount of inflammatory cytokines after oral administration of the present invention maple extract to mice with colitis induced by 5% DSS.
Figure 6 is an image showing the expression of ZO-1 increased after oral administration of colitis induction of colitis induced by 5% DSS of the present invention, and Western blot image showing the expression of ZO-1 and ocludine.
이하 본 발명을 실시예를 통하여 보다 상세하게 설명한다. 그러나 이들 실시예는 본 발명을 예시적으로 설명하기 위한 것으로 본 발명의 범위가 이들 실시예에 한정되는 것은 아니다.Hereinafter, the present invention will be described in more detail with reference to Examples. However, these examples are for illustrative purposes only and the scope of the present invention is not limited to these examples.
제조예 1: 단풍나무 에탄올 추출물(KIOM-2015E)의 제조 Preparation Example 1 Preparation of Maple Ethanol Extract (KIOM-2015E)
단풍나무 잎 분쇄물 1700g을 17ℓ의 25% 에탄올에 넣고 24시간 실온 침적하여 추출하였다. 그 후, 상기 추출물을 감압농축하고 동결건조하여 단풍나무 잎 25% 에탄올 추출물 78.57g을 수득하였다.1700 g of maple leaf pulverized product was put in 17 L of 25% ethanol and extracted by immersion at room temperature for 24 hours. Thereafter, the extract was concentrated under reduced pressure and lyophilized to obtain 78.57 g of maple leaf 25% ethanol extract.
실시예 1: 염증성 장 질환을 유도한 마우스의 체중 측정Example 1 Body Weight Measurement of Mice Inducing Inflammatory Bowel Disease
C57BL/6 마우스에 염증성 장 질환을 유도하기 위하여 5% DSS(Dextran sodium sulfate)를 6일 동안 급수하였다. 마우스에 생리식염수 또는 생리식염수에 녹인 KIOM-2015E (100 mg/kg) 또는 5-Amino-salacylic acid (5-ASA, 100 mg/kg)를 9일 동안 경구 투여하였으며, 7일째에 5% DSS를 물로 바꾸어 주었다.C57BL / 6 mice were watered with 5% Dexran sodium sulfate (DSS) for 6 days to induce inflammatory bowel disease. Mice were given oral administration of KIOM-2015E (100 mg / kg) or 5-Amino-salacylic acid (5-ASA, 100 mg / kg) dissolved in saline or saline for 9 days, and 5% DSS on
염증성 장 질환을 유도한 마우스의 체중을 측정한 결과 도 1과 같이, 물을 급수한 마우스에 비해서 염증성 장 질환이 유도된 5% DSS 투여 마우스에서는 체중 감소가 나타나는 것을 확인할 수 있었다. 반면 5% DSS를 투여하며 KIOM-2015E 100mg/kg, 5-ASA 100mg/kg을 경구 투여한 마우스에서는 체중 감소가 상대적으로 경미하게 일어나는 것을 확인할 수 있었다.As a result of measuring the weight of the mouse induced inflammatory bowel disease, as shown in Figure 1, it was confirmed that the weight loss in 5% DSS-induced mice induced inflammatory bowel disease compared to water-watered mice. On the other hand, mice receiving oral administration of KIOM-2015E 100mg / kg and 5-ASA 100mg / kg with 5% DSS were found to have relatively mild weight loss.
이는, 염증성 장 질환에 의해 소화 장애가 생기면서 유발되는 체중 감소를 단풍나무 추출물에 의해 완화시킨 것으로 판단할 수 있다.It can be determined that the weight loss caused by digestive disorders caused by inflammatory bowel disease is alleviated by maple extract.
실시예 2: 염증성 장 질환을 유도한 마우스의 대장 길이 측정Example 2: Measurement of colon length in mice induced with inflammatory bowel disease
상기 실시예 1에서 염증성 장 질환을 유도한 마우스의 대장을 적출하여 대장 길이를 측정하였다.In Example 1, the colon of the mice inducing inflammatory bowel disease was extracted and the colon length was measured.
그 결과 도 2과 같이, 물을 급수한 마우스에 비해서 염증성 장 질환이 유도된 5% DSS 투여 마우스는 매우 짧은 대장의 길이를 갖는 것을 확인하였다. 반면 5% DSS를 투여하며 KIOM-2015E 100mg/kg, 5-ASA 100mg/kg을 경구 투여한 마우스에서는 장 질환 유도 마우스와 비교하여 대장의 길이 감소가 완화된 것을 확인할 수 있었다.As a result, as shown in Figure 2, compared to the water-induced mice, 5% DSS-induced inflammatory bowel disease-induced mice were confirmed to have a very short colon length. On the other hand, mice that received oral administration of KIOM-2015E 100mg / kg and 5-ASA 100mg / kg with 5% DSS were found to have reduced bowel length reduction compared to intestinal disease-induced mice.
이는, 장 질환에 의해 유발되는 증상 중 하나인 대장의 길이 감소를 단풍나무 추출물이 저해함으로써 단풍나무 추출물이 장 질환 관련 증상을 억제할 수 있음을 시사한다.This suggests that maple extract may inhibit bowel disease related symptoms by inhibiting the length reduction of the large intestine, which is one of the symptoms caused by bowel disease.
실시예 3: 염증성 장 질환을 유도한 마우스의 대장 조직 병변 확인 Example 3: Confirmation of Colon Tissue Lesions in Mice Inducing Inflammatory Bowel Disease
상기 실시예 1에서 염증성 장 질환을 유도한 마우스의 대장을 내시경을 통해 병변을 확인하였으며, 적출한 대장 조직을 파라포름알데하이드로 고정 시킨 후, H&E, Alcian blue, PAS 염색을 통하여 대장 조직 병변을 확인하였다.In Example 1, the lesion of the mouse inducing inflammatory bowel disease was confirmed through an endoscope, and the colon tissue was confirmed by H & E, Alcian blue, and PAS staining after fixing the extracted colon tissue with paraformaldehyde. It was.
조직 병변을 확인한 결과, 도 3과 같이, 물을 급수한 마우스에서는 대장 조직이 정상인 것을 확인한 반면, 5% DSS 투여 마우스에서는 대장 조직에 염증이 많이 발생한 것을 확인하였다. 5% DSS를 투여하며 KIOM-2015E 100mg/kg 및 5-ASA 100mg/kg을 경구 투여한 마우스에서는 조직에 염증이 감소되어 있는 것을 확인하였다.As a result of confirming the tissue lesion, it was confirmed that the colon tissue is normal in the water-watered mice, as shown in Figure 3, while in the 5% DSS-administered mice it was confirmed that a lot of inflammation in the colon tissue. Mice administered with 5% DSS and orally administered KIOM-
이는, 단풍나무 추출물이 장내 염증반응을 억제하여 염증성 장 질환을 치료할 수 있음을 시사한다.This suggests that maple extract can treat inflammatory bowel disease by inhibiting the intestinal inflammatory response.
실시예 4: 염증성 장 질환을 유도한 마우스의 대식세포의 침투 확인Example 4 Confirmation of Macrophage Penetration of Mice Inducing Inflammatory Bowel Disease
상기 실시예 1에서 염증성 장 질환을 유도한 마우스에 luciferase를 포함하는 대식세포를 tail vein injection을 통해 침투시킨 후, in vivo image system으로 대식세포의 침투를 확인하였으며 대장 조직에서 F4-80을 염색한 후 대식세포의 침투를 확인하였다.After infiltrating macrophage containing luciferase into the mouse induced inflammatory bowel disease in Example 1 through tail vein injection, the macrophage infiltration was confirmed using an in vivo image system and stained with F4-80 in colon tissue. After the penetration of macrophages was confirmed.
대식세포의 침투를 확인한 결과, 도 4와 같이 물을 급수한 마우스에서는 대식세포의 침투가 적은 반면, 5% DSS 투여 마우스에서는 대식세포의 침투가 많이 발생한 것을 확인하였다. 5% DSS를 투여하며 KIOM-2015E 100mg/kg 및 5-ASA 100mg/kg을 경구 투여한 마우스에서는 대식세포의 침투가 감소되어 있는 것을 확인하였다.As a result of confirming the infiltration of macrophages, as shown in Figure 4, the water-induced mice, while the macrophages are less invasive, it was confirmed that 5% DSS-induced macrophage infiltration was large. It was confirmed that macrophage infiltration was reduced in mice administered 5% DSS and orally administered KIOM-2015E 100mg / kg and 5-ASA 100mg / kg.
이는, 단풍나무 추출물이 염증 반응에 관여하는 대식세포의 침투를 감소시킴으로써 대장 조직 내 염증반응을 억제할 수 있음을 시사한다.This suggests that maple extract can inhibit the inflammatory response in colon tissue by reducing the infiltration of macrophages involved in the inflammatory response.
실시예 5: 염증성 장 질환을 유도한 마우스의 염증성 사이토카인 분비 확인Example 5: Confirmation of Inflammatory Cytokine Secretion in Mice Inducing Inflammatory Bowel Disease
상기 실시예 1에서 염증성 장 질환을 유도한 마우스의 혈액을 채취하여 염증성 사이토카인인 Interleukin(IL)-6 및 Tumor necrosis factor (TNF)-α의 분비양을 확인하였다.In Example 1, blood was collected from mice inducing inflammatory bowel disease to confirm the amount of inflammatory cytokines Interleukin (IL) -6 and Tumor necrosis factor (TNF) -α.
염증성 사이토카인의 분비양을 확인한 결과, 도 5와 같이 물을 급수한 마우스에서는 염증성 사이토카인의 분비양이 적은 반면, 5% DSS 투여 마우스에서는 염증성 사이토카인의 분비양이 증가한 것을 확인하였다. 5% DSS를 투여하며 KIOM-2015E 100mg/kg 및 5-ASA 100mg/kg을 경구 투여한 마우스에서는 염증성 사이토카인의 분비양이 감소되어 있는 것을 확인하였다. As a result of confirming the secretion amount of inflammatory cytokines, it was confirmed that the amount of inflammatory cytokines secreted in the water-watered mice as shown in Figure 5, while the amount of inflammatory cytokines increased in 5% DSS-treated mice. Mice administered with 5% DSS and orally administered KIOM-
이는, 단풍나무 추출물이 염증반응에 직접적으로 관여하는 염증성 사이토카인의 분비를 감소시킴으로써 염증반응을 억제하는 효과가 있음을 시사한다.This suggests that maple extract has an effect of inhibiting the inflammatory response by reducing the secretion of inflammatory cytokines directly involved in the inflammatory response.
실시예 6: 염증성 장 질환을 유도한 마우스의 장내 밀착 연접 보호효과Example 6: Intestinal tight splice protection effect of mice induced inflammatory bowel disease
상기 실시예 1에서 염증성 장 질환을 유도한 마우스의 대장 조직을 파라포름알데하이드로 고정시킨 후, ZO-1 1차 항체를 이용하여 대장조직내 발현되는 ZO-1의 양을 측정하였으며, 또한 대장 조직으로부터 단백질을 추출하여 ZO-1 및 오클루딘의 발현양을 웨스턴 블롯법으로 확인하였다.In Example 1, the colon tissue of the mouse induced inflammatory bowel disease was fixed with paraformaldehyde, and then the amount of ZO-1 expressed in colon tissue was measured using a ZO-1 primary antibody. The protein was extracted from and the expression amounts of ZO-1 and ocludine were confirmed by Western blot.
밀착 연접 단백질인 ZO-1 및 오클루딘의 발현양을 확인한 결과, 도 6과 같이 5% DSS 투여 마우스에서 ZO-1 및 오클루딘의 발현양의 감소로 밀착 연접이 손상되어 있는 것을 확인하였다. 반면, 5% DSS를 투여하며 KIOM-2015E 100mg/kg 및 5-ASA 100mg/kg을 경구 투여한 마우스에서는 ZO-1 및 오클루딘의 발현양의 증가로 밀착 연접이 회복되어 있는 것을 확인하였다.As a result of confirming the expression levels of the tight junction protein ZO-1 and occlusin, as shown in Figure 6 it was confirmed that in the 5% DSS-administered mice, the tight junction is damaged by the decrease in the expression amount of ZO-1 and occlusin. On the other hand, mice that received oral administration of KIOM-
이는, 단풍나무 추출물이 염증성 장 질환에 의해 유발되는 증상 중 하나인 밀착 연접의 기능 장애를 회복하는 효과를 가져 장 질환에 대한 치료 효과가 있음을 시사한다.This suggests that the maple extract has a therapeutic effect on intestinal diseases because it has an effect of restoring dysfunction of the close junction, which is one of the symptoms caused by inflammatory bowel disease.
이상의 설명으로부터, 본 발명이 속하는 기술분야의 당업자는 본 발명이 그 기술적 사상이나 필수적 특징을 변경하지 않고서 다른 구체적인 형태로 실시될 수 있다는 것을 이해할 수 있을 것이다. 이와 관련하여, 이상에서 기술한 실시 예들은 모든 면에서 예시적인 것이며 한정적인 것이 아닌 것으로서 이해해야만 한다. 본 발명의 범위는 상기 상세한 설명보다는 후술하는 특허 청구범위의 의미 및 범위 그리고 그 등가 개념으로부터 도출되는 모든 변경 또는 변형된 형태가 본 발명의 범위에 포함되는 것으로 해석되어야 한다.From the above description, those skilled in the art will appreciate that the present invention can be implemented in other specific forms without changing the technical spirit or essential features. In this regard, the embodiments described above are to be understood in all respects as illustrative and not restrictive. The scope of the present invention should be construed that all changes or modifications derived from the meaning and scope of the following claims and equivalent concepts rather than the detailed description are included in the scope of the present invention.
Claims (7)
Pharmaceutical composition for preventing or treating intestinal diseases, including maple leaf extract.
상기 추출물은 물, 탄소수 1내지 4의 알코올 또는 이들의 혼합용매로 추출한 것인, 약학 조성물.
The method of claim 2,
The extract is extracted with water, alcohol having 1 to 4 carbon or a mixed solvent thereof, pharmaceutical composition.
상기 장 질환은 과민성 장 증후군 또는 염증성 장 질환인 것인, 약학 조성물.
The method of claim 2,
The bowel disease is irritable bowel syndrome or inflammatory bowel disease, pharmaceutical composition.
A method of preventing or treating intestinal diseases comprising administering maple leaf extract to a subject other than a human.
Health functional food for the prevention or improvement of intestinal diseases comprising maple leaf extract.
Feed composition for preventing or improving intestinal diseases, including maple leaf extract.
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