KR102487701B1 - Centella asiatica extract for bronchitis or respiratory disease and composition comprising the same as an active ingredient and use thereof - Google Patents
Centella asiatica extract for bronchitis or respiratory disease and composition comprising the same as an active ingredient and use thereof Download PDFInfo
- Publication number
- KR102487701B1 KR102487701B1 KR1020220057064A KR20220057064A KR102487701B1 KR 102487701 B1 KR102487701 B1 KR 102487701B1 KR 1020220057064 A KR1020220057064 A KR 1020220057064A KR 20220057064 A KR20220057064 A KR 20220057064A KR 102487701 B1 KR102487701 B1 KR 102487701B1
- Authority
- KR
- South Korea
- Prior art keywords
- bronchitis
- centella asiatica
- asthma
- extract
- ethyl acetate
- Prior art date
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Abstract
본 발명은 기관지염 또는 호흡기 질환 개선효능을 가지는 병풀 추출물에 관한것으로서, 보다 자세하게는, 병풀을 수집한 후, 병풀 건초를 열수, 에탄올 추출한 추출물을 이용하여, 기관지염 또는 호흡기 질환의 지표인, 점액질 및 β-hexosaminidase활성에 대한 억제를 확인하고, 그 중 가장 효과가 우수한 병풀 추출물을 기관지염 또는 호흡기 질환 개선, 예방 및 치료용 조성물에 유효성분으로서 포함될 수 있는 천연물에 관한 것이다.The present invention relates to a centella asiatica extract having an effect of improving bronchitis or respiratory diseases, and more particularly, after collecting centella asiatica, using an extract obtained by extracting centella hay with hot water and ethanol, mucus and β, indicators of bronchitis or respiratory diseases, are obtained. - Confirmation of inhibition of hexosaminidase activity, and relates to a natural product that can be included as an active ingredient in a composition for improving, preventing, and treating bronchitis or respiratory disease with centella asiatica extract, which is the most effective among them.
Description
본 발명은 기관지염 또는 호흡기 질환 개선효능을 갖는 병풀 추출물에 관한 것으로서, 더욱 상세하게는 병풀 추출물을 확보하여, MUC5AC와 히스타민 분비와 연관된 β-hexosaminidaes 억제활성을 확인함으로써, 천식, 만성폐쇄성폐질환, 급만성기관지염, 알레르기비염, 기침, 가래, 인후염, 편도염, 후두염과 같은 급성 상기도 감염증 등의 호흡기 질환의 예방 또는 치료에 유용한 조성물을 제공한다.The present invention relates to a centella asiatica extract having bronchitis or respiratory disease improving efficacy, and more particularly, by securing the centella asiatica extract and confirming the β-hexosaminidaes inhibitory activity associated with MUC5AC and histamine secretion, thereby treating asthma, chronic obstructive pulmonary disease, acute Provided is a composition useful for preventing or treating respiratory diseases such as chronic bronchitis, allergic rhinitis, cough, sputum, sore throat, tonsillitis, and acute upper respiratory infections such as laryngitis.
천식, 만성폐쇄성폐질환, 알레르기비염, 기침, 가래, 급만성기관지염, 세기관지염, 인후염, 편도염, 후두염 등은 호흡기질환의 대표적인 염증성 질환이다.Asthma, chronic obstructive pulmonary disease, allergic rhinitis, cough, sputum, acute and chronic bronchitis, bronchiolitis, sore throat, tonsillitis, laryngitis, etc. are representative inflammatory diseases of respiratory diseases.
대표적인 호흡기 질환인 천식(Asthma)은 만성적인 기도 염증에 의한 비특이적 기도과민성 및 가역적 기도 폐쇄를 특징으로 하는 질환이다. 호흡 곤란, 기침, 색색거리는 거친 숨소리(천명) 등의 증상이 반복적으로 발작적으로 나타나는 병증으로, 천식 환자의 약 30%에서 생후 1년 이내에 약 80%에서는 4~5세에 처음 천식 증상을 나타내며 우리나라에서도 약 10% 내외의 발병률을 나타내고 있다. 천식을 병태/생리학적으로 보면, 기도 염증, 기도 과민성, 점막단백 과다분비가 일어나는 질환이며, 면역학적으로는 호산구의 침윤, Th2 세포수의 증가, 활성화 비만 세포 수가 증가되는 것을 특징으로 하는 만성기도 염증 질환이다.Asthma, a representative respiratory disease, is a disease characterized by non-specific airway hyperresponsiveness and reversible airway obstruction due to chronic airway inflammation. It is a disease in which symptoms such as dyspnoea, coughing, and wheezing are repeated and paroxysmal. About 30% of asthma patients and about 80% within 1 year of age have asthma symptoms for the first time at 4 to 5 years of age, and in Korea It also shows an incidence rate of about 10%. From a pathological/physiological point of view, asthma is a disease in which airway inflammation, airway hyperresponsiveness, and mucosal protein hypersecretion occur. Immunologically, it is a chronic airway characterized by eosinophil infiltration, an increase in the number of Th2 cells, and an increase in the number of activated mast cells. It is an inflammatory disease.
천식은 네 가지 병적 증상으로 요약될 수 있는데, 기도 내의 호산구의 유입이 현저하게 증가하고, 점액이 과다하게 분비되며, 부종이 관찰되기도 하고, 무엇보다 기도가 좁아지는 것을 특징적인 소견으로 하고 있다.Asthma can be summarized into four pathological symptoms, which include markedly increased influx of eosinophils into the airways, excessive secretion of mucus, edema, and, above all, narrowing of the airways.
호흡기도는 크게 점막과 기관지 평활근이라는 근육으로 이루어져 있고 점막에는 많은 분비샘들이 있어 필요한 분비물을 계속 분비하고 있으며 기관지 평활근이 수축하면 호흡기도가 좁아지게 된다. 매연, 알레르기성 항원, 찬바람, 운동, 호흡기 감염 등 매우 다양한 소인에 의하여 염증반응이 일어나면 분비샘에서 나오는 분비물이 더욱 증가하게 되고 이 분비물이 기도를 막아 점막이 기도 안쪽으로 부어오르게 되어 기도를 더욱 좁게 만든다. 이로 인해, 천명을 동반한 발작적인 기침과 호흡곤란이 심하게 나타나며, 발작 시에는 마른기침이 발생되고 흉부압박감을 느끼게 된다. 천명이 없이 만성적 기침과 흉부압박감이 있는 원인을 알 수 없는 호흡곤란 증상만 있는 천식도 많은데 이 증상들은 일상생활 중에서 갑자기 발작적으로 나타나는 경향이 있다.The respiratory tract is largely composed of a mucous membrane and a muscle called bronchial smooth muscle, and there are many secretory glands in the mucous membrane to continuously secrete necessary secretions. When the bronchial smooth muscle contracts, the respiratory airway narrows. When an inflammatory reaction occurs due to a wide variety of factors such as smoke, allergens, cold wind, exercise, and respiratory infections, the secretion from the glands further increases, and this secretion blocks the airway, causing the mucous membrane to swell into the airway, further narrowing the airway. . As a result, paroxysmal coughing accompanied by wheezing and difficulty in breathing appear severely, and during seizures, dry cough occurs and chest pressure is felt. There are many asthmatic patients who have only symptoms of chronic cough and chest tightness without wheezing and unexplained dyspnea, but these symptoms tend to appear suddenly and paroxysmal during daily life.
현재, 천식은 기관지 협착증에서 만성적인 기관지 염증질환으로 개념이 새로 정립되었으며, 증상이 있을 때 이를 완화시키는 것도 중요하지만 장기적으로 염증을 관리하는 근본적인 치료방법이 중요하다.Currently, the concept of asthma has been newly established as a chronic bronchial inflammatory disease in bronchial stenosis, and it is important to alleviate symptoms when there are symptoms, but a fundamental treatment method to manage inflammation in the long term is important.
천식을 병태/생리학적 원인으로 보면, 기도염증(airway inflammation), 기도과민성(airway-hyper responsiveness, AHR), 점막단백 과다분비(mucin hyper secretion)가 일어나는 질환이며, 면역학적으로는 호산구(eosinophils)의 침윤, Th1 세포수에 비해 Th2 세포의 수가 증가하거나, 활성화 비만 세포(mast cell)수가 증가되는 것을 특징으로 하는 만성기도 염증질환이다. 천식의 증상으로는 기도에 백혈구 중 과립구(granulocyte) 계통인 호산구가 침윤되는 것이 대표적인 특징으로, 호산구는 기도염증과 기관지 수축을 촉진하는 여러 유발물질을 생성하여 천식의 병태/생리에 매우 중요한 역할을 수행한다. 천식을 일으키는 항원은 T 세포를 Th2 세포로의 분화를 일으키며, Th2 세포는 IL(interleukin)-5, GM-CSF(granulocyte-macrophage stimulating factor), IL-3, IL-13, IL-4 등의 사이토카인(cytokine)을 분비하고, IL-4는 B 세포에 작용하여 IgE의 생성을 촉진시키며, 비만세포를 활성화 시킨다. 활성화된 비만세포는, 탈과립 되어 히스타민이 분비되어지며, 이때 β-hexosamindase도 함께 분비되어 진다. 이런 효소적 활성은 비만세포 탈과립의 척도로 사용될 수 있어 실험에 사용할 수 있다. 비만세포가 활성화 되어 탈과립화 되면, 다양한 염증 매개인자를 유리하며, 기도에서 기관지수축, 혈관확장, 감각신경 감작화, 콜린성 기관지 수축 증의 급성 염증반응을 일으키게 된다. 대부분의 천식은 가역적이지만 일부 환자의 경우 천식이 진행됨에 따라 상치하세포의 섬유화, 혈관수 및 점액분비 세포수의 증가 및 기도 평활근의 비후에 의한 기도 구조변화로 리모델링이 일어나며, 만성폐쇄성질환(chronic obstructive pulmonary disease, COPD)에서 나타나는 기도폐쇄도 일어날 수 있다.Looking at the pathological/physiological causes of asthma, it is a disease in which airway inflammation, airway-hyper responsiveness (AHR), and mucin hyper secretion occur. Immunologically, eosinophils It is a chronic airway inflammatory disease characterized by infiltration, an increase in the number of Th2 cells compared to the number of Th1 cells, or an increase in the number of activated mast cells. A typical symptom of asthma is the infiltration of eosinophils, a type of granulocyte among white blood cells, into the airways. Eosinophils play a very important role in the pathophysiology/physiology of asthma by producing various triggers that promote airway inflammation and bronchial constriction. carry out Asthma-inducing antigen causes differentiation of T cells into Th2 cells, and Th2 cells produce IL-5, GM-CSF (granulocyte-macrophage stimulating factor), IL-3, IL-13, IL-4, etc. It secretes cytokines, and IL-4 acts on B cells to promote the production of IgE and activate mast cells. Activated mast cells degranulate and secrete histamine, at which time β-hexosamindase is also secreted. This enzymatic activity can be used as a measure of mast cell degranulation and can be used in experiments. When mast cells are activated and degranulated, they release various inflammatory mediators and cause an acute inflammatory response in the airway, such as bronchoconstriction, vasodilation, sensory nerve sensitization, and cholinergic bronchoconstriction. Most asthma is reversible, but in some patients, remodeling occurs due to structural changes in the airway due to fibrosis of subdental cells, increase in the number of blood vessels and mucus-secreting cells, and thickening of airway smooth muscle as asthma progresses. Airway obstruction as seen in obstructive pulmonary disease (COPD) may also occur.
만성폐쇄성질환은 원인이 되는 폐질환이나 심장질환이 없이 기도폐쇄가 발생하여 기류의 속도가 감소하는 질환군을 일컫는다. 임상적으로는 만성적으로 객담을 동반하는 기침을 하는 만성기관지염과 종말세기관지 이하의 폐포들이 비정상적으로 늘어나고 폐포격벽이 파괴되는 폐기종이 혼합되어 구분이 힘든 경우에, 이들을 총칭하여 만성폐쇄성폐질환이라고 한다. 만성폐쇄성폐질환은 천식과 비슷하게 호흡곤란, 기침, 가래 등의 기도 질환증상을 나타내다가 폐 기능을 악화시켜 결국에는 사망에 이르게 된다. 발병의 주요 원인은 흡연으로, 이외에도 공해와 선천적 질환, 호흡기 감염증 등이 원인이 되며, 호중성구(neutrophils) 및 대식세포(macrophage)에 의해 유도된다고 알려져 있다. 호중성구는 대표적인 염증세포로서 여러 종류의 단백분해효소를 분비하여 폐실질의 파과와 만성적인 점액분비를 일으키며, 대식세포는 최근 IFN-γ(interferon-γ), IL-13 과발현의 중요한 염증세포로 대두되고 있고, 이는 조직손상을 일으키는 매개체(reactive oxygen species, ROS 대사산물 등)를 분비할 뿐만 아니라 상처치유에 관여하는 매개체(transforming growth factor, TGF-β; fibroblast growth factor, FGF2; endothelial growth factor, VEGF 등)를 분비하는 만성염증을 일으키는 주요원인이 되는 세포이다. 비가역적 기도폐색은 폐포의 파괴로 인해 주변 기도가 막혀 발생하는 폐기종과, 소기도인 세기관지의 염증과 이에 따른 반복적인 손상에 의해 섬유화를 특징으로 발생하는 만성폐쇄성세기관지염(small airway fibrosis, obstructive bronchiolitis)이 있다. 이러한 질환의 환자들에게는 대부분 폐기종과 함께 소기도의 염증과 섬유화가 동반하여 나타난다.Chronic obstructive disease refers to a group of diseases in which airflow velocity decreases due to airway obstruction without a causative lung disease or heart disease. Clinically, when chronic bronchitis with chronic cough accompanied by sputum and emphysema in which alveoli below the terminal bronchioles are abnormally enlarged and the alveolar septum is destroyed, it is difficult to distinguish them, collectively referred to as chronic obstructive pulmonary disease. Chronic obstructive pulmonary disease (COPD) presents airway disease symptoms such as dyspnea, cough, and phlegm, similar to asthma, but worsens lung function and eventually leads to death. The main cause of the outbreak is smoking, in addition to pollution, congenital diseases, respiratory infections, etc., and is known to be induced by neutrophils and macrophages. Neutrophils are representative inflammatory cells that secrete various types of proteolytic enzymes to cause breakthrough of the lung parenchyma and chronic secretion of mucus. emerging, which not only secretes mediators that cause tissue damage (reactive oxygen species, ROS metabolites, etc.) but also mediators involved in wound healing (transforming growth factor, TGF-β; fibroblast growth factor, FGF2; endothelial growth factor, It is a cell that secretes VEGF, etc.) and is the main cause of chronic inflammation. Irreversible airway obstruction is characterized by emphysema, which occurs when the surrounding airways are blocked due to destruction of alveoli, and chronic obstructive bronchiolitis, which is characterized by fibrosis caused by inflammation of the bronchioles, which are small airways, and consequent repeated damage. there is Most patients with these diseases present with inflammation and fibrosis of the small airways along with emphysema.
알레르기 비염(allergic rhinitis)은 알레르기 염증 반응으로 인하여 콧물, 재채기, 코 및 눈 가려움증 등의 증상이 나타나는 질환으로 삶의 질을 악화시키는 질환이다. 대부분의 산업국가에서는 인구의 8~24%가 이러한 질환을 나타낼 정도로 성인과 소아에서 매우 흔한 만성 질환 중의 하나이다. 비염은 알레르기가 아닌 다른 기전에 의해서도 나타날 수 있으나 알레르기 기전이 전체 비염의 약 2/3를 차지할 정도로 알레르기가 비염 발생의 중요한 요인이다. 알레르기 비염은 천식과 흔히 동반되며 유전적인 경향과 병태생리를 많이 공유한다. Allergic rhinitis is a disease in which symptoms such as runny nose, sneezing, itching in the nose and eyes appear due to an allergic inflammatory reaction, and it is a disease that deteriorates the quality of life. In most industrialized countries, it is one of the most common chronic diseases in adults and children, with 8-24% of the population exhibiting this disease. Rhinitis can also be caused by mechanisms other than allergy, but allergy is an important factor in the occurrence of rhinitis to the extent that allergic mechanisms account for about 2/3 of all rhinitis. Allergic rhinitis often accompanies asthma and shares many genetic predispositions and pathophysiology.
진해는 기침을 그치게 하는 것을 말하며, 거담은 가래를 없애는 것을 말한다. 찬 공기, 병원성 미생물을 포함한 외부 이물질, 대기 오염 물질, 알레르기 유발 물질 등과 같은 물리 화학적 요인 등에 의해 기침, 가래가 발생될 수 있다. 기침은 기도 점막 자극에 의해 반사적으로 일어나는 방어기전으로, 지나친 자극에 의한 지속적인 기침이 유발될 경우, 환자의 삶의 질을 경감시키고 악화시키게 된다. 또한 기침과 같은 원리로 외부에서 먼지나 자극 물질 등이 유입되면 우리 몸의 기관지에서는 타액과 함께 근육운동을 해서 외부로 밀어내게 되는데, 이것이 객담(가래) 형성의 원인이며 폐 등 기관지 염증에 의해서 짙은 화농성 객담이 생기게 된다.Jinhae means to stop coughing, and expectoration means to get rid of phlegm. Cough and phlegm may be caused by physical and chemical factors such as cold air, external foreign substances including pathogenic microorganisms, air pollutants, and allergens. Coughing is a defense mechanism that occurs reflexively by irritation of the airway mucosa, and when persistent coughing is induced by excessive stimulation, the patient's quality of life is reduced or deteriorated. In addition, when dust or irritants are introduced from the outside on the same principle as coughing, the bronchi of our body exert muscle movement along with saliva to push them out. Purulent sputum is produced.
급성기관지염(acute bronchitis)은 기관과 기관지의 급성 염증으로 대개는 호흡계의 감염과 관련이 있다. 급성 기관지염은 원인에 따라 급성 감염성 기관지염과 급성 자극성 기관지염으로 분류할 수 있는데 감염성 기관지염의 원인은 대부분 바이러스로 상기도 감염을 동반한다. 급성 자극성 기관지염은 다양한 금속성 물질이나 휘발성 용매, 이산화질소나 담배 등의 물질에 의한 자극, 혹은 알레르겐을 흡입함으로써 나타나는 알레르기 반응 등에 의한 것일 수 있다. 만성기관지염(chronic bronchitis)은 일반적으로 1년에 3개월 이상 2년 연속적으로 거의 매일 기침 및 과도한 가래가 나오는 만성 기관지점막의 염증으로 정의하며, 만성 기관지염 환자는 기침의 증가, 가래의 양과 화농성의 증가, 호흡 곤란 등 빈번하게 급성 악화를 경험하게 된다. 여러 국가에서 만성 기관지염과 그 악화로 인한 사망률이 증가하고 있다.Acute bronchitis is an acute inflammation of the trachea and bronchi, usually associated with infection of the respiratory system. Acute bronchitis can be classified into acute infectious bronchitis and acute irritant bronchitis depending on the cause. Most of the causes of infectious bronchitis are viruses, which are accompanied by upper respiratory infections. Acute irritant bronchitis may be caused by irritation by substances such as various metal substances, volatile solvents, nitrogen dioxide or tobacco, or allergic reactions caused by inhaling allergens. Chronic bronchitis is generally defined as inflammation of the bronchial mucosa with coughing and excessive sputum production almost every day for at least 3 months per year for 2 consecutive years. , shortness of breath, and frequent acute exacerbations. Death rates from chronic bronchitis and its exacerbation are increasing in many countries.
현재 이 같은 호흡기 질환에 있어서의 근본적인 치료법은 없으며 발작을 예방하고 합병증을 방지하기 위한 여러 가지 방법과 약물이 쓰이나 만족할 만한 효과를 얻지 못하는 실정이다. 주로 흡입용 기관지 확장제, 경구용 또는 주사용 기관지 확장제(교감신경 자극제 및 테오필린계 약제들), 스테로이드 제제(흡입용, 경구용, 주사용 등), 류코트리엔 길항제(몬테루카스트, 트란루카스트, 질루론 등), 항알러지 약제(디소디움 클로로클리케이트, 크로몰린 소디움, 케토티펜)등이 치료제로 활용되고 있다. 항콜린제, 베타2 수용체 항진제 등의 기도 확장제는 질병을 악화시키는 염증에는 효과가 없어 단순한 증상만 완화하기 때문에 장기간 사용 시 약제 내성 발생 및 병증 악화의 우려가 있다. 염증에 효과가 있다고 알려진 스테로이드제는 심각한 부작용으로 인한 장기간 사용 시 문제가 되고 있다. 따라서, 이 둘을 병용 처방하는 경우가 많은데 스테로이드류의 부작용으로 인해 경구제보다 흡입제 형태로 개발되어 복용이 어렵기 때문에 복용 순응도가 떨어지는 문제점이 있다.Currently, there is no fundamental treatment for such respiratory diseases, and various methods and drugs are used to prevent seizures and complications, but satisfactory effects are not obtained. Mainly inhaled bronchodilators, oral or injectable bronchodilators (sympathetic nerve stimulants and theophylline drugs), steroids (inhalation, oral, injection, etc.), leukotriene antagonists (Montelukast, Tranlukast, Zyluron, etc.) ), anti-allergy drugs (disodium chloroclicate, cromolyn sodium, ketotifen) are being used as treatments. Airway dilators, such as anticholinergics and beta2 receptor agonists, are ineffective against inflammation that exacerbates the disease and only relieve simple symptoms, so there is concern about drug resistance and worsening of symptoms when used for a long time. Steroids, which are known to be effective for inflammation, are problematic when used for a long time due to serious side effects. Therefore, in many cases, the two are prescribed together, but due to the side effects of steroids, it is developed in the form of an inhalant rather than an oral agent, so it is difficult to take it, so there is a problem with poor dosage compliance.
따라서, 이러한 현 약물 치료제의 한계점을 극복하여, 원인을 근본적으로 치료하고 증상을 효과적으로 개선할 수 있는 새로운 치료제의 개발이 필요하다.Therefore, it is necessary to develop a new therapeutic agent capable of fundamentally treating the cause and effectively improving symptoms by overcoming the limitations of these current drug therapeutic agents.
따라서, 상기와 같은 문제점을 해결하기 위하여 안출된 본 발명의 목적은, 병풀 추출물을 유효성분으로 포함하는 기관지염 또는 호흡기 질환의 예방 또는 치료용 약학적 조성물을 제공하는 것이다.Therefore, an object of the present invention, which was made to solve the above problems, is to provide a pharmaceutical composition for preventing or treating bronchitis or respiratory diseases containing centella asiatica extract as an active ingredient.
본 발명의 다른 목적은, 병풀 추출물을 유효성분으로 포함하는 기관지염 또는 호흡기 질환의 예방 또는 개선용 식품 조성물을 제공하는 것이다.Another object of the present invention is to provide a food composition for preventing or improving bronchitis or respiratory diseases, containing centella asiatica extract as an active ingredient.
본 발명의 또 다른 목적은, 병풀(Centella asiatica)을 물, C1 내지 C4의 저급 알코올 또는 이들의 혼합 용매로 추출하는 단계; 및 상기 추출물을 헥산, 부탄올, 에틸아세테이트 및 증류수 중 선택된 하나의 용매로 분획하는 단계；를 포함하는 기관지염 또는 호흡기 질환의 예방, 개선 또는 치료용 조성물의 제조 방법을 제공하는 것이다.Another object of the present invention is to extract centella asiatica with water, C1 to C4 lower alcohol or a mixed solvent thereof; And fractionating the extract with one solvent selected from hexane, butanol, ethyl acetate and distilled water; to provide a method for preparing a composition for preventing, improving or treating bronchitis or respiratory diseases, including a.
전술한 목적을 달성하기 위해 본 발명에서는, 병풀 추출물을 유효성분으로 포함하는 기관지염 또는 호흡기 질환의 예방 또는 치료용 약학적 조성물을 제공한다.In order to achieve the above object, the present invention provides a pharmaceutical composition for the prevention or treatment of bronchitis or respiratory diseases, containing centella asiatica extract as an active ingredient.
상기의 병풀 추출물은 기관지염 또는 호흡기 질환인 천식, 만성폐쇄성폐질환, 알레르기성 비염, 기관지염, 감기 또는 기침인 것을 특징으로 하는 기관지염 또는 호흡기 질환 예방 또는 치료용 약학적 조성물을 제공한다.The Centella Asiatica extract provides a pharmaceutical composition for preventing or treating bronchitis or respiratory disease, characterized in that bronchitis or respiratory disease asthma, chronic obstructive pulmonary disease, allergic rhinitis, bronchitis, cold or cough.
상기 천식은 알레르기성 천식, 비알레르기성 천식, 아토피성 천식 및 아토피성 기관지 IgE 매개 천식으로 이루어진 군에서 1이상 선택되는 것을 특징으로 한다.The asthma is characterized in that at least one selected from the group consisting of allergic asthma, non-allergic asthma, atopic asthma and atopic bronchial IgE-mediated asthma.
상기 기관지염은, 만성 기관지염, 급성 기관지염, 카타르성 기관지염, 세기관지염 또는 폐쇄성 및 염증성 기관지 질병으로 이루어진 군에서 1이상 선택되는 것을 특징으로 한다.The bronchitis is characterized in that at least one selected from the group consisting of chronic bronchitis, acute bronchitis, catarrhal bronchitis, bronchiolitis or obstructive and inflammatory bronchial diseases.
본 발명에서의 상기 병풀 추출물은, 열수, C1 내지 C4의 저급 알콜 및 10~95 부피%의 저급 알콜 수용액 추출물인 것을 특징으로 한다.The Centella asiatica extract in the present invention is characterized in that it is an extract of hot water, C1 to C4 lower alcohol and 10 to 95% by volume of lower alcohol aqueous solution.
상기 마다가스카르산 병풀 추출물은 70 부피% 에탄올 수용액 추출물인 것을 특징으로 한다.The Madagascar Centella asiatica extract is characterized in that it is an extract of 70% by volume ethanol aqueous solution.
상기 병풀 추출물은 70 부피% 에탄올 수용액 추출물에서, 헥산, 에틸 아세테이트, 부탄올 및 증류수로 용매 분획된 것을 더 포함할 수 있다.The Centella Asiatica extract may further include solvent fractionation with hexane, ethyl acetate, butanol, and distilled water in a 70% ethanol aqueous solution extract.
또한, 본 발명에서는 상기 병풀 추출물을 유효성분으로 포함하는 기관지염 또는 호흡기 질환 예방 또는 개선용 식품 조성물을 제공한다.In addition, the present invention provides a food composition for preventing or improving bronchitis or respiratory diseases comprising the centella asiatica extract as an active ingredient.
상기의 병풀 추출물은 기관지염 또는 호흡기 질환인 천식, 만성폐쇄성폐질환, 알레르기성 비염, 기관지염, 감기 또는 기침인 것을 특징으로 하는 기관지염 또는 호흡기 질환 예방 또는 개선용 식품 조성물을 제공한다.The Centella Asiatica extract provides a food composition for preventing or improving bronchitis or respiratory diseases, characterized in that bronchitis or respiratory diseases such as asthma, chronic obstructive pulmonary disease, allergic rhinitis, bronchitis, cold or cough.
상기 천식은 알레르기성 천식, 비알레르기성 천식, 아토피성 천식 및 아토피성 기관지 IgE 매개 천식으로 이루어진 군에서 1이상 선택되는 것을 특징으로 한다.The asthma is characterized in that at least one selected from the group consisting of allergic asthma, non-allergic asthma, atopic asthma and atopic bronchial IgE-mediated asthma.
상기 기관지염은, 만성 기관지염, 급성 기관지염, 카타르성 기관지염, 세기관지염 또는 폐쇄성 및 염증성 기관지 질병으로 이루어진 군에서 1이상 선택되는 것을 특징으로 한다.The bronchitis is characterized in that at least one selected from the group consisting of chronic bronchitis, acute bronchitis, catarrhal bronchitis, bronchiolitis or obstructive and inflammatory bronchial diseases.
본 발명에서의 상기 병풀 추출물은, 열수, C1 내지 C4의 저급 알콜 및 10~95 부피%의 저급 알콜 수용액 추출물인 것을 특징으로 한다.The Centella asiatica extract in the present invention is characterized in that it is an extract of hot water, C1 to C4 lower alcohol and 10 to 95% by volume of lower alcohol aqueous solution.
상기 병풀 추출물은 70 부피% 에탄올 수용액 추출물인 것을 특징으로 한다.The Centella asiatica extract is characterized in that it is an extract of 70% by volume ethanol aqueous solution.
상기 병풀 추출물은 70 부피% 에탄올 수용액 추출물에서, 헥산, 에틸 아세테이트, 부탄올 및 증류수로 용매 분획된 것을 더 포함할 수 있다.The Centella Asiatica extract may further include solvent fractionation with hexane, ethyl acetate, butanol, and distilled water in a 70% ethanol aqueous solution extract.
또한, 본 발명에서는 병풀(Centella asiatica)을 물, C1 내지 C4의 저급 알코올 또는 이들의 혼합 용매로 추출하는 단계; 및 상기 추출물을 헥산, 부탄올, 에틸아세테이트 및 증류수 중 선택된 하나의 용매로 분획하는 단계；를 포함하는 기관지염 또는 호흡기 질환의 예방, 개선 또는 치료용 조성물의 제조 방법을 제공한다.In the present invention, extracting centella asiatica with water, C1 to C4 lower alcohol or a mixed solvent thereof; And fractionating the extract with one solvent selected from hexane, butanol, ethyl acetate and distilled water; It provides a method for preparing a composition for preventing, improving or treating bronchitis or respiratory diseases, including a.
전술한 바와 같은 본 발명의 병풀 추출물은, 점액질 및 베타-헥소사미니다제(β-hexosaminidase)와 같은 알러지 유발인자를 억제하는 활성을 가지고 있어, 이를 이용하여 기관지염 또는 호흡기 질환의 개선을 위한 다양한 조성물의 유효성분으로 사용할 수 있으며, 부작용을 가지는 합성의약품을 대체하여, 기관지염 또는 호흡기 질환의 예방용 식품조성물, 치료용 약학적 조성물로 사용할 수 있다.As described above, the centella asiatica extract of the present invention has an activity to inhibit mucus and allergens such as beta-hexosaminidase, and various compositions for improving bronchitis or respiratory diseases using the same It can be used as an active ingredient, and can be used as a food composition for preventing bronchitis or respiratory diseases, and a pharmaceutical composition for treatment, replacing synthetic drugs having side effects.
도 1은 본 발명의 병풀 추출물을 추출하는 방법을 모식화한 도이다. 건조된 병풀은 열수, 70% 에탄올, 95% 에탄올로 추출되었으며, 70% 에탄올 추출물은 추가적으로 헥산, 에틸 아세테이트, 부탄올, 증류수로 추가 분획하였다.
도 2는 본 발명의 용매에 따라 추출된 병풀 추출물의 NCI-H292세포에 대한, 추출 용매별 세포독성을 확인한 결과를 나타낸 도이다.
도 3은 본 발명의 NCI-H292세포를 PMA로 자극하여 분비된 점액질(MUC5AC)에 대하여, 용매별 병풀 추출물의 점액 분비 억제 활성을 평가한 도이다.
도 4는 본 발명의 NCI-H292세포를 PMA로 자극하여 분비된 점액질(MUC5AC)에 대하여, 용매별 병풀 추출물의 점액 생성 억제 활성을 평가한 도이다.
도 5는 본 발명의 병풀을 70%에탄올로 추출한 후 추가로 용매 분획한 분획물을 이용하여, NCI-H292세포에 대한 세포독성을 측정한 결과를 나타낸 도이다.
도 6은 본 발명의 NCI-H292세포에서, PMA로 자극하여 병풀 추출물 및 추가 용매 분획물을 이용한 점액질(MUC5AC)의 분비 억제 활성을 평가한 도이다.
도 7은 본 발명의 병풀 추출물 및 추가 용매 분획물을, IgE로 자극된 RBL-2H3세포에 처리하여, β-hexosaminidase의 억제 활성을 평가한 도이다.1 is a diagram schematically illustrating a method for extracting the centella asiatica extract of the present invention. The dried centella asiatica was extracted with hot water, 70% ethanol, and 95% ethanol, and the 70% ethanol extract was additionally fractionated with hexane, ethyl acetate, butanol, and distilled water.
Figure 2 is a diagram showing the results of confirming the cytotoxicity of each extraction solvent for NCI-H292 cells of the centella asiatica extract extracted according to the solvent of the present invention.
Figure 3 is a view showing the mucus secretion inhibitory activity of Centella Asiatica extracts for each solvent with respect to mucin (MUC5AC) secreted by stimulating NCI-H292 cells with PMA of the present invention.
Figure 4 is a view showing the mucin production inhibitory activity of centella asiatica extracts for each solvent with respect to mucin (MUC5AC) secreted by stimulating NCI-H292 cells with PMA of the present invention.
Figure 5 is a diagram showing the results of measuring cytotoxicity to NCI-H292 cells using a fraction obtained by extracting centella asiatica of the present invention with 70% ethanol and further solvent-fractionating it.
Figure 6 is a diagram showing the secretion inhibitory activity of mucilage (MUC5AC) using centella asiatica extract and additional solvent fractions stimulated with PMA in NCI-H292 cells of the present invention.
7 is a diagram illustrating the evaluation of inhibitory activity of β-hexosaminidase by treating IgE-stimulated RBL-2H3 cells with centella asiatica extract and additional solvent fractions of the present invention.
이하, 본 발명에서 사용한 용어를 설명한다.Hereinafter, terms used in the present invention will be described.
본 발명에서 사용되는 용어 “추출물”이란 약제 또는 식물체내의 광합성 또는 대사산물을 공지의 용매를 이용하여 수득한 산물을 의미하며, 복합물을 의미한다.The term "extract" used in the present invention refers to a product obtained from photosynthesis or metabolites in a drug or plant using a known solvent, and refers to a complex.
본 발명에서 사용되는 용어 “분획물”이란, 혼합물 혹은 복합물에 있어서, 복합물의 극성 또는 비극성, 수용성 또는 불용성 등과 같은 친화도에 따라, 공지의 용매를 사용하여 성상별로 분류하여 수득한 것을 의미한다. The term "fraction" used in the present invention means obtained by classifying a mixture or composite according to its properties using a known solvent according to affinity such as polarity or nonpolarity, water solubility or insolubility of the composite.
본 발명에서 “약제학적으로 허용가능한”이란 생물체를 상당히 자극하지 않고 투여 활성 물질의 생물학적 활성 및 특성을 저해하지 않는 것을 의미한다.In the present invention, "pharmaceutically acceptable" means that it does not significantly stimulate living organisms and does not inhibit the biological activity and properties of the active substance administered.
본 발명에서 사용되는 용어 “예방”은 본 발명의 조성물의 투여로 특정 질환의 증상을 억제하거나 진행을 지연시키는 모든 행위를 의미한다.As used herein, the term "prevention" refers to any action that suppresses symptoms or delays the progression of a specific disease by administering the composition of the present invention.
본 발명에서 사용되는 용어 “치료”는 본 발명의 조성물의 투여로 특정 질환의 증상을 호전 또는 이롭게 변경시키는 모든 행위를 의미한다.The term "treatment" used in the present invention refers to all activities that improve or beneficially change the symptoms of a specific disease by administering the composition of the present invention.
본 발명에서 사용되는 용어 “개선”은 치료되는 상태와 관련된 파라미터, 예를 들면 증상의 정도를 적어도 감소시키는 모든 행위를 의미한다.As used herein, the term "improvement" refers to any action that at least reduces the severity of a parameter related to the condition being treated, for example, a symptom.
이하, 본 발명을 상세히 설명한다.Hereinafter, the present invention will be described in detail.
본 발명은 기관지염 또는 호흡기 질환 개선효능을 갖는 병풀 추출물을 제공한다.The present invention provides a centella asiatica extract having bronchitis or respiratory disease improving efficacy.
상기 본 발명의 병풀(Centella asiatica)은 인도, 남아프리카, 마다가스카르, 한국, 일본 및 중국등의 열대 및 온대 지역에서 자생하는 식물로서, 전초를 약용하는 식물이며, 다양한 종류의 플라보노이드(flavonoid) 배당체가 함유되어 있다. 한방에서는 청열, 이습, 소종, 해독에 효과가 있다고 알려져 있으며, 상업적으로는 피부손상 및 피부질환의 개선뿐만 아니라, 염증 및 궤양억제, 항산화, 항균을 위한 성분으로 사용되어 지고 있다.Centella asiatica of the present invention is a plant that grows wild in tropical and temperate regions such as India, South Africa, Madagascar, Korea, Japan and China, and is a medicinal herb, and contains various types of flavonoid glycosides has been In oriental medicine, it is known to be effective in clearing heat, dampness, sore throat, and detoxification, and commercially, it is used as a component for not only improving skin damage and skin diseases, but also inhibiting inflammation and ulcers, antioxidant, and antibacterial.
본 발명에서는 상기의 병풀을 산지별로 수집하였으며, 건조된 병풀을 열수, 70% 에탄올, 95% 에탄올로 추출하여 수득된 추출물을 수득하였으며, 추출물 중 기관지염 또는 호흡기 질환에 가장 효과적인 70%에탄올 추출물을 헥산, 에틸 아세테이트, 부탄올 및 증류수로 분획하여, 세포독성 및 기관지염 또는 호흡기 질환에 대한 개선효과를 평가하였고, 그 결과 마다가스카르산 병풀 추출물 또는 분획물이 효과가 가장 우수함을 확인하였다.In the present invention, the centella asiatica was collected by production area, and an extract obtained by extracting the dried centella asiatica with hot water, 70% ethanol, and 95% ethanol was obtained, and among the extracts, the most effective 70% ethanol extract for bronchitis or respiratory diseases was hexane , fractionated with ethyl acetate, butanol and distilled water to evaluate the improvement effect on cytotoxicity and bronchitis or respiratory diseases, and as a result, it was confirmed that the Madagascar Centella asiatica extract or fraction had the most excellent effect.
또한, 본 발명은 상기의 병풀 추출물 또는 용매 분획물을 유효성분으로 함유하는 조성물을 제공한다.In addition, the present invention provides a composition containing the Centella asiatica extract or solvent fraction as an active ingredient.
본 발명의 조성물은 통상적인 약제 추출물의 제조방법에 따라 제조될 수 있으며, 일반적으로, 액상이나 건조분말 형태일 수 있다. 또한 주 성분인 상기 병풀 추출물 또는 용매 분획물의 유효량에 1종 또는 2종 이상의 약제학적으로 허용 가능한 통상적인 담체 또는 1종 또는 2종 이상의 첨가제를 선택하여 통상적인 제형의 조성물로 제조할 수 있다.The composition of the present invention may be prepared according to a conventional method for preparing a drug extract, and may generally be in the form of a liquid or dry powder. In addition, one or two or more pharmaceutically acceptable conventional carriers or one or two or more additives may be selected in an effective amount of the Centella asiatica extract or solvent fraction as a main component to prepare a composition in a conventional dosage form.
담체는 희석제, 활택제, 결합제, 붕해제, 감미제, 안정제, 방부제 중에서 1종 또는 2종 이상을 선택하여 사용할 수 있으며, 첨가제로는 향료, 비타민류, 항산화제 중에서 1종 또는 2종 이상을 선택하여 사용할 수 있다.One or two or more carriers may be selected from diluents, lubricants, binders, disintegrants, sweeteners, stabilizers, and preservatives, and one or two or more additives may be selected from among flavoring agents, vitamins, and antioxidants. and can be used.
본 발명에 있어서, 담체 및 첨가제는 그 종류에 제한되는 것은 아니며, 구체적으로는 희석제로 유당(lactose monohydrate), 트레할로스(Trehalose), 옥수수 전분(corn starch), 콩기름(soybean oil), 미결정 셀룰로오스(microcrystalline cellulose) 또는 만니톨(D-mannitorl), 활택제로는 스테아린산 마그네슘(magnesiumstearate) 또는 탈크(talc), 결합제로는 폴리비닐피롤리돈(PVP:polyvinyipyrolidone) 또는 하이드록시프로필셀룰로오스(HPC: hydroxypropylcellulose) 중에서 선택할 수 있다. 또한, 붕해제로는 카르복시메틸셀룰로오스칼슘(Ca-CMC: carboxymethylcellulose calcium), 전분글리콜산나트륨(sodium starchglycolate), 폴라크릴린칼륨(polacrylin potassium) 또는 크로스포비돈(cross-linkedpolyvinylpyrrolidone), 감미제로는 백당, 과당, 소르비톨(sorbitol) 또는 아스파탐(aspartame), 안정제로는 카르복시메틸셀룰로오스나트륨(Na-CMC: carboxymethylcellulosesodium), 베타-싸이크로덱스트린(β-cyclodextrin), 백납(white bee's wax) 또는 잔탄검(xanthan gum) 중에서 선택할 수 있으며, 방부제로는 파라옥시안식향산메칠(methyl p-hydroxy benzoate, methlparaben), 파라옥시안식향산프로필(propyl p-hydroxybenzoate, propylparaben), 또는 소르빈산칼륨(potassium sorbate) 중에서 선택할 수 있다.In the present invention, carriers and additives are not limited to their types, and specifically, lactose monohydrate, trehalose, corn starch, soybean oil, and microcrystalline cellulose as diluents cellulose) or mannitol (D-mannitorl), magnesium stearate or talc as a lubricant, and polyvinylpyrrolidone (PVP: polyvinyipyrolidone) or hydroxypropylcellulose (HPC) as a binder. there is. In addition, disintegrants include carboxymethylcellulose calcium (Ca-CMC), sodium starch glycolate, polacrylin potassium or cross-linked polyvinylpyrrolidone, and sweeteners include sucrose, Fructose, sorbitol or aspartame, carboxymethylcellulosesodium (Na-CMC) as a stabilizer, β-cyclodextrin, white bee's wax or xanthan gum ), and the preservative may be selected from methyl p-hydroxy benzoate (methlparaben), propyl p-hydroxybenzoate (propylparaben), or potassium sorbate.
또한, 본 발명은 상기 병풀 추출물 또는 용매 분획물을 유효성분으로 함유하는 약학적 조성물, 건강 기능식품 조성물 또는 일반 식품 조성물을 제공한다.In addition, the present invention provides a pharmaceutical composition, health functional food composition or general food composition containing the centella asiatica extract or solvent fraction as an active ingredient.
상기 병풀 추출물에는 식물체가 생산하는 염증 활성 억제 물질이 포함되어 있다. The Centella Asiatica extract contains substances that inhibit inflammation activity produced by plants.
또한, 상기 본 발명에 따른 조성물은 본 발명의 병풀 추출물과 함께 섭취하기 적합하고, 섭취 시 기관지염 또는 호흡기 질환을 개선시키는 다른 종류의 공지된 추출물 및 약제를 추가로 포함할 수 있다.In addition, the composition according to the present invention is suitable for ingestion with the Centella Asiatica extract of the present invention, and may further include other types of known extracts and drugs that improve bronchitis or respiratory diseases when ingested.
본 발명의 상기 약학적 조성물은 기관지염 또는 호흡기 질환에 대한 예방 또는 치료 효과를 포함한다. 본 발명의 병풀 추출물 또는 용매 분획물은, 앞서 언급한 바, 점액 분비 및 염증 인자의 활성을 억제 효과를 가지므로, 이러한 효과를 통하여, 본 발명의 마다가스카르산 병풀 추출물 또는 용매 분획물은 기관지염 또는 호흡기 질환에 대한 예방 또는 치료 효과를 가질 수 있다. 상기 기관지염 또는 호흡기 질환은 이에 제한되는 것은 아니나, 천식, 알레르기성 비염, 기관지염, 만성 폐쇄성 질환 감기 또는 기침 중에서 선택되는 1 이상의 질환을 의미한다. 본 발명의 상기 기관지염 또는 호흡기 질환의 유발인자인 점액질의 분비 억제, β-hexosaminidase 활성 억제를 통하여 기관지염 또는 호흡기 질환의 개선 효과를 확인하였다. The pharmaceutical composition of the present invention includes a preventive or therapeutic effect on bronchitis or respiratory diseases. As mentioned above, the Centella asiatica extract or solvent fraction of the present invention has an effect of inhibiting mucus secretion and the activity of inflammatory factors, so through these effects, the Centella asiatica extract or solvent fraction of the present invention is effective in treating bronchitis or respiratory diseases. may have a preventive or therapeutic effect on The bronchitis or respiratory disease refers to one or more diseases selected from asthma, allergic rhinitis, bronchitis, chronic obstructive disease, cold, or cough, but is not limited thereto. The improvement effect of bronchitis or respiratory diseases was confirmed through inhibition of mucus secretion and inhibition of β-hexosaminidase activity, which are inducing factors of bronchitis or respiratory diseases of the present invention.
본 발명의 건강기능식품은 병풀 추출물 또는 용매 분획물을 유효성분으로 함유하는 차, 젤리, 즙, 엑기스, 음료 등을 포함할 수 있으며, 상기 검토한 기능을 포함하고 있는 것이라면 가공 형태에 제한이 있는 것은 아니다.The health functional food of the present invention may include tea, jelly, juice, extract, beverage, etc. containing centella asiatica extract or solvent fraction as an active ingredient, and if it contains the functions discussed above, there are restrictions on the processing form not.
또한, 본 발명의 상기 식품은 건강기능식품과 구별되는 일반 식품으로서, 아이스크림류, 우유, 두유, 치즈를 포함하는 유제품, 및 두유제품, 음료, 육류, 소세지, 빵, 초콜릿 캔디류, 스낵류, 과자류, 피자, 라면, 기타 면류, 껌류, 각종 수프, 음료수, 차, 드링크제, 알코올 음료, 캡슐 또는 스틱 포장의 유산균 제제로 구성된 군으로부터 선택할 수 있다.In addition, the food of the present invention is a general food that is distinguished from health functional food, and includes ice cream, milk, soy milk, dairy products including cheese, and soy milk products, beverages, meat, sausages, bread, chocolate candies, snacks, confectionery, It may be selected from the group consisting of pizza, ramen, other noodles, chewing gum, various soups, beverages, tea, drinks, alcoholic beverages, and lactic acid bacteria preparations in capsule or stick packaging.
본 발명의 병풀 추출물 또는 용매 분획물은, 점액질 분비 및 활성 억제능이 우수하고, 염증 유발 인자의 생성 억제 능력이 뛰어나므로, 상기 마다가스카르산 병풀 추출물 또는 용매 분획물은 기관지염 또는 호흡기 질환 개선을 위한 약학적 조성물, 건강기능 식품 조성물 또는 일반 식품 조성물 등 다양한 형태로 활용될 수 있다. 또한, 상기 기능을 활용할 수 있는 비말제, 유제품 및 식품 등의 첨가제로서 제한 없이 활용될 수 있다.Since the Centella asiatica extract or solvent fraction of the present invention has excellent mucus secretion and activity inhibition ability and excellent ability to inhibit the production of inflammatory factors, the Madagascar Centella asiatica extract or solvent fraction is a pharmaceutical composition for improving bronchitis or respiratory disease, It can be used in various forms such as health functional food compositions or general food compositions. In addition, it can be used without limitation as an additive for splash agents, dairy products, and foods that can utilize the above function.
상기 식품에 대한 일 례로서, 본 발명의 병풀 추출물 또는 용매 분획물을 포함하는 음료 조성물의 경우 통상의 음료와 같이 여러 가지 향미제 또는 천연 탄수화물 등을 추가 성분으로서 함유할 수 있다. 상술한 천연 탄수화물은 포도당, 과당과 같은 모노사카라이드, 말토오스 슈크로오스와 같은 디사카라이드, 및 덱스트린, 사이클로덱스트린과 같은 폴리사카라이드, 자일리톨, 소르비톨, 에리트리톨 등의 당알코올이다. 감미제로서는 타우마틴, 스테비아 추출물과 같은 천연 감미제나 사카린, 아스파르탐과 같은 합성 감미제 등을 사용할 수 있다.As an example for the food, in the case of a beverage composition containing the centella asiatica extract or solvent fraction of the present invention, various flavoring agents or natural carbohydrates may be included as additional components, like conventional beverages. The aforementioned natural carbohydrates include monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As the sweetener, natural sweeteners such as thaumatin and stevia extract or synthetic sweeteners such as saccharin and aspartame may be used.
상기 외의 본 발명의 병풀 추출물 또는 용매 분획물을 포함하는 조성물은 여러 가지 영양제, 비타민, 전해질, 풍미제, 착색제, 펙트산 및 그의 염, 알긴산 및 그의 염, 유기산, 보호성 콜로이드 증점제, pH 조절제, 안정화제, 방부제, 글리세린, 알코올, 탄산음료에 사용되는 탄산화제 등을 함유할 수 있다. 이러한 성분은 독립적으로 또는 조합하여 사용할 수 있다.In addition to the above, the composition containing the centella asiatica extract or solvent fraction of the present invention contains various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH regulators, and stability agents. It may contain topical agents, preservatives, glycerin, alcohol, carbonation agents used in carbonated beverages, and the like. These components may be used independently or in combination.
아울러 상기 본 발명의 병풀 추출물 또는 이의 용매 분획물을 포함하는 조성물은 성분의 함량에 있어 크게 제한되는 것은 아니며, 본 발명에서 목적하는 기능을 타나낼 수 있는 양으로 포함할 수 있다. 즉, 유효성분의 혼합양은 사용 목적(예방, 건강 또는 치료적 처치)에 따라 적합하게 결정될 수 있다.In addition, the composition containing the centella asiatica extract or a solvent fraction thereof of the present invention is not significantly limited in the content of the component, and may be included in an amount capable of exhibiting the desired function in the present invention. That is, the mixing amount of the active ingredient may be appropriately determined according to the purpose of use (prevention, health or therapeutic treatment).
또한 본 발명은, 병풀(Centella asiatica)을 물, C1 내지 C4의 저급 알코올 또는 이들의 혼합 용매로 추출하는 단계; 및 상기 추출물을 헥산, 부탄올, 에틸아세테이트 및 증류수 중 선택된 하나의 용매로 분획하는 단계；를 포함하는 기관지염 또는 호흡기 질환의 예방, 개선 또는 치료용 조성물의 제조 방법을 제공한다.In addition, the present invention, extracting Centella asiatica with water, C1 to C4 lower alcohol or a mixed solvent thereof; And fractionating the extract with one solvent selected from hexane, butanol, ethyl acetate and distilled water; It provides a method for preparing a composition for preventing, improving or treating bronchitis or respiratory diseases, including a.
이하, 본 발명의 실시예를 첨부된 도면을 참고하여 보다 상세하게 설명하도록 한다. 그러나, 하기의 실시예는 본 발명의 내용을 구체화하기 위한 것일 뿐, 이에 의해 본 발명이 한정되는 것은 아닐 것이다.Hereinafter, embodiments of the present invention will be described in more detail with reference to the accompanying drawings. However, the following examples are only for specifying the contents of the present invention, and the present invention will not be limited thereby.
<실시예 1> 마다가스카르산 병풀 추출물 제조방법<Example 1> Manufacturing method of centella asiatica extract from Madagascar
병풀에는 피부질환 개선 효능 및 염증, 궤양억제, 항암, 면역 증진 등 다양한 생리활성이 존재하는 것으로 확인되어 있으나, 기관지 및 호흡기 질환에 대한 개선 효과는 보고된 바가 없다. 따라서, 본 발명에서는 다양한 산지별 마다가스카르산 병풀 추출물을 확보하여 호흡기 건강용 기능성 소재로서의 활용 가능성을 타진하고자 하였다. 먼저 마다가스카르산 병풀을 건초형태로 수집하였으며, 수집된 건초는 열수, 70% 에탄올 및 95% 에탄올로 추출되었다(도 1). 열수 추출물은, 증류수의 20 중량%로 병풀 건초를 첨가한 후, 90 ℃에서 2시간동안 중탕하여, 추출물을 수득하였다. 70% 에탄올 추출물은 70% 에탄올의 10 중량%로 병풀 건초를 첨가한 후 80℃에서 2시간 동안 환류 추출 하였으며, 95% 에탄올 추출 또한 상기 70% 에탄올 추출과 같은 방법으로 추출하였다. 추출된 각각의 용매 추출물은 여과한 후 건조시켜, 동결건조를 통해 분말화 하였다.Centella asiatica has been confirmed to have various physiological activities such as skin disease improvement efficacy, inflammation, ulcer inhibition, anticancer, and immunity enhancement, but the improvement effect on bronchial and respiratory diseases has not been reported. Therefore, in the present invention, the possibility of utilization as a functional material for respiratory health was sought by securing extracts of Centella Asiatica from Madagascar for various production regions. First, Centella Asiatica from Madagascar was collected in the form of hay, and the collected hay was extracted with hot water, 70% ethanol and 95% ethanol (FIG. 1). For the hot water extract, centella hay was added in 20% by weight of distilled water, and then heated in hot water at 90 ° C. for 2 hours to obtain an extract. The 70% ethanol extract was refluxed for 2 hours at 80 ° C. after adding centella hay at 10% by weight of 70% ethanol, and 95% ethanol extraction was also extracted in the same way as the 70% ethanol extraction. Each extracted solvent extract was filtered, dried, and powdered through lyophilization.
<실시예 2> 마다가스카르산 병풀 추출물 활성 평가<Example 2> Evaluation of activity of Centella Asiatica extract from Madagascar
2-1. 기관지 세포에 대한 세포독성 평가2-1. Evaluation of cytotoxicity to bronchial cells
본 발명에서 추출된 용매별 마다가스카르산 병풀 추출물은, 96 well plate에 배양된 기관지 세포 NCI-H292에 농도별로 처리하여 세포독성을 평가하였다. 처리된 농도는 62.5 μg/ml, 125 μg/ml, 250 μg/ml, 500 μg/ml, 1000 μg/ml 농도로 처리하였으며, 대조군으로는 PMA를 단독 처리하여 비교하였다. 그 결과, 70% 에탄올 추출물인 CA-70E, 열수추출물(CA-95HW) 및 95% 에탄올 추출물(CA-95E)은 유의적인 독성을 나타내지 않았다(도 2).Centella asiatica extract from Madagascar for each solvent extracted in the present invention was treated with bronchial cells NCI-H292 cultured in a 96 well plate for each concentration to evaluate cytotoxicity. The treated concentrations were treated at concentrations of 62.5 μg/ml, 125 μg/ml, 250 μg/ml, 500 μg/ml, and 1000 μg/ml, and PMA was treated alone as a control and compared. As a result, 70% ethanol extract CA-70E, hot water extract (CA-95HW) and 95% ethanol extract (CA-95E) did not show significant toxicity (FIG. 2).
2-2. 점액 분비 및 생성 억제활성 평가2-2. Evaluation of mucus secretion and production inhibitory activity
기관지염 또는 호흡기질환의 주요 증상인 점액질의 분비 및 생성억제활성을 평가하기 위하여, NCI-H292 기관지 상피세포에서 포르볼 12-미리스테이트 13-아세테이트(phorbol 12-myristate 13-acetate, PMA)를 처리하여, MUC5AC 분비(secretion) 및 생성(production)을 유도한 후, 실험예 2-1에서 사용한 병풀 추출물의 처리농도와 동일하게 처리하여, 점액질 분비 및 생성 억제를 평가하였다.In order to evaluate the secretion and production inhibitory activity of mucus, which is a major symptom of bronchitis or respiratory disease, NCI-H292 bronchial epithelial cells were treated with phorbol 12-myristate 13-acetate (PMA) , MUC5AC secretion and production were induced, and then mucus secretion and production inhibition were evaluated by treating with the same treatment concentration as the centella asiatica extract used in Experimental Example 2-1.
점액질 분비 억제는, 기관지 상피세포에서, CA-70E가 62.5~1000 μg/ml의 농도범위에서 15.7~100.2%의 억제 활성을 나타내어, CA-HW(0.7~10.5% 억제) 및 CA-95E(2.3~26.4% 억제)에 비해 우수한 활성을 보여주었다(도 3).In bronchial epithelial cells, CA-70E exhibited 15.7-100.2% inhibitory activity in the concentration range of 62.5-1000 μg/ml, and CA-HW (0.7-10.5% inhibition) and CA-95E (2.3%) inhibited mucous secretion. ~26.4% inhibition) showed superior activity (Fig. 3).
점액질 생성 억제 또한 점액질 분비 억제와 유사한 결과를 보여주었으며, CA-70E가 6.8~79.8%, CA-95E 가 8.5~27.2%, CA-HW가 2.9~7.8% 순으로 점액질 생성을 억제하였다(도 4). Inhibition of mucus production also showed similar results to inhibition of mucus secretion, CA-70E inhibited mucus production in the order of 6.8 to 79.8%, CA-95E by 8.5 to 27.2%, and CA-HW by 2.9 to 7.8% (FIG. 4). ).
<실험예 3> 마다가스카르산 병풀 분획물 및 활성 평가<Experimental Example 3> Madagascar centella asiatica fraction and activity evaluation
3-1. 마다가스카르산 병풀 분획물 제조 및 세포독성 평가3-1. Preparation of centella asiatica fraction from Madagascar and evaluation of cytotoxicity
상기 실시예 2에서 확인된 병풀 추출물 CA-70E에 대한 각 분획물의 세포 독성을 평가하였다. The cytotoxicity of each fraction against the centella asiatica extract CA-70E identified in Example 2 was evaluated.
구체적으로, CA-70E 80 부피%를 분획 용매로서 각각 헥산, 에틸아세테이트, 부탄올 또는 증류수 20 부피%와 혼합한 후 강하게 볼텍싱하여 층 분리 하였으며, 분리된 층을 취해 건조시켜 각각 CA-70E-Hex, CA-70E-EtOAc, CA-70E-BuOH 및 CA-70E-Water로 분말화 하여 제조하였다(도 1).Specifically, 80% by volume of CA-70E was mixed with 20% by volume of hexane, ethyl acetate, butanol or distilled water as fractionation solvents, and then the layers were separated by strong vortexing, and the separated layers were taken and dried to obtain CA-70E-Hex , prepared by powdering with CA-70E-EtOAc, CA-70E-BuOH and CA-70E-Water (FIG. 1).
추가로 분획된 병풀 분획물은 마다가스카르산 병풀 추출물의 처리농도(실시예 2-1)와 동일한 농도로 NCI-H292세포에 처리되어, 세포독성을 평가하였다. 대조군으로, PMA 처리군과 CA-70E를 처리한 군을 함께 배양하여 비교하였다. 그 결과를 도 5에 나타내었다. 각 분획물은 세포 독성이 확인되지 않음을 확인하였다. The additionally fractionated Centella asiatica fraction was treated with NCI-H292 cells at the same concentration as the treatment concentration of the Centella asiatica extract from Madagascar (Example 2-1), and cytotoxicity was evaluated. As a control group, the PMA-treated group and the CA-70E-treated group were cultured together and compared. The results are shown in FIG. 5 . It was confirmed that each fraction had no cytotoxicity.
3-2. 병풀 분획물에 대한 점액질 분비 억제 및 알러지 인자 억제 활성 평가3-2. Evaluation of mucus secretion inhibition and allergy factor inhibition activity for centella asiatica fractions
CA-70E를 추가로 분획한 병풀 분획물의 활성을 평가하였으며, 점액질에 대한 억제 활성을 도 6에 나타내었다. 100 μg/ml의 농도로 병풀 분획물을 PMA로 자극된 NCI-H292 세포에 처리하여, MUC5AC의 분비 억제를 평가한 결과, CA-70E(49% 억제)에 비해 용매분획 된 CA-70E-EtOAc는 MUC5AC를 81.5% 억제한 것을 확인 할 수 있고, CA-70E-BuOH 또한 용매분획을 통해 점액질 분비억제가 증진되었음을 확인하였다.The activity of the centella asiatica fraction, in which CA-70E was additionally fractionated, was evaluated, and the inhibitory activity against mucilage was shown in FIG. 6 . The centella asiatica fraction was treated with PMA-stimulated NCI-H292 cells at a concentration of 100 μg/ml to evaluate the inhibition of MUC5AC secretion. It was confirmed that MUC5AC was inhibited by 81.5%, and CA-70E-BuOH was also confirmed to enhance mucous secretion inhibition through solvent fractionation.
또한, IgE로 자극된 기관지 세포인 RBL-2H3을 이용하여, 알러지 유발인자인 β-hexosaminidase의 억제활성을 평가하였다. 점액질 분비 억제시 처리했던 동일 농도로, 용매 분획물을 처리하여, 확인한 결과 CA-70E는 43.9%의 β-hexosaminidase 억제능을 보였으며, CA-70E-EtOAc는 69.7%억제하였으며, CA-70E-BuOH 또한 용매분획을 통해 β-hexosaminidase의 활성도 억제된 것을 확인하였다.In addition, the inhibitory activity of β-hexosaminidase, an allergen, was evaluated using IgE-stimulated bronchial cells, RBL-2H3. As a result of processing the solvent fraction at the same concentration used when inhibiting mucous secretion, it was confirmed that CA-70E inhibited β-hexosaminidase by 43.9%, CA-70E-EtOAc by 69.7%, and CA-70E-BuOH also inhibited β-hexosaminidase by 43.9%. It was confirmed that the activity of β-hexosaminidase was also inhibited through solvent fractionation.
Claims (6)
상기 기관지염 또는 호흡기 질환은 천식, 만성폐쇄성폐질환, 알레르기성 비염, 기관지염, 감기 또는 기침인 것을 특징으로 하는, 기관지염 또는 호흡기 질환 예방 또는 치료용 약학적 조성물. Centella asiatica 70% by volume ethanol aqueous solution extract or its ethyl acetate fraction containing as an active ingredient, mucus secretion and production inhibition and beta-hexosaminidase (β-hexosaminidase) through inhibition of activity bronchitis or As a pharmaceutical composition for the prevention or treatment of respiratory diseases,
The bronchitis or respiratory disease is characterized in that asthma, chronic obstructive pulmonary disease, allergic rhinitis, bronchitis, cold or cough, bronchitis or respiratory disease prevention or treatment pharmaceutical composition.
상기 기관지염 또는 호흡기 질환은 천식, 만성폐쇄성폐질환, 알레르기성 비염, 기관지염, 감기 또는 기침인 것을 특징으로 하는, 기관지염 또는 호흡기 질환 예방 또는 개선용 식품 조성물. Centella asiatica 70% by volume ethanol aqueous solution extract or its ethyl acetate fraction containing as an active ingredient, mucus secretion and production inhibition and beta-hexosaminidase (β-hexosaminidase) through inhibition of activity bronchitis or As a food composition for preventing or improving respiratory diseases,
The bronchitis or respiratory disease is characterized in that asthma, chronic obstructive pulmonary disease, allergic rhinitis, bronchitis, cold or cough, bronchitis or respiratory disease prevention or improvement food composition.
상기 추출물을 헥산, 부탄올, 에틸아세테이트 및 증류수로 분획하여 에틸아세테이트 분획물을 얻는 단계；를 포함하는,
제1항의 기관지염 또는 호흡기 질환의 예방, 개선 또는 치료용 조성물의 제조 방법.
Extracting Centella asiatica with a 70% by volume ethanol aqueous solution; and
Fractionating the extract with hexane, butanol, ethyl acetate and distilled water to obtain an ethyl acetate fraction;
A method for preparing a composition for preventing, improving or treating bronchitis or respiratory diseases of claim 1.
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