KR102642516B1 - Composition for preventing, improving or treating respiratory diseases comprising mixture of Ecklonia cava extract etc as an active ingredient and method of manufacturing the same - Google Patents
Composition for preventing, improving or treating respiratory diseases comprising mixture of Ecklonia cava extract etc as an active ingredient and method of manufacturing the same Download PDFInfo
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- KR102642516B1 KR102642516B1 KR1020220187312A KR20220187312A KR102642516B1 KR 102642516 B1 KR102642516 B1 KR 102642516B1 KR 1020220187312 A KR1020220187312 A KR 1020220187312A KR 20220187312 A KR20220187312 A KR 20220187312A KR 102642516 B1 KR102642516 B1 KR 102642516B1
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- South Korea
- Prior art keywords
- ecklonia cava
- chrysanthemum
- extract
- cava extract
- preventing
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Classifications
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/105—Plant extracts, their artificial duplicates or their derivatives
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Abstract
본 발명은 감태추출물 및 감국추출물을 함유하는 감태추출물혼합물을 유효성분으로 포함하는 호흡기 질환 예방 또는 개선용 건강기능식품 조성물, 및 감태추출물 및 감국추출물을 함유하는 감태추출물혼합물을 유효성분으로 포함하는 호흡기 질환 예방 또는 치료용 약학적 조성물, 및 감태추출물혼합물의 제조방법에 관한 것으로, 뮤신 분비의 증가에 의해 발생 또는 악화되거나 뮤신 분비가 증가하는 증상을 동반하는 호흡기 질환의 예방, 개선 또는 치료에 우수한 효능을 나타낼 수 있다.The present invention provides a health functional food composition for preventing or improving respiratory diseases containing an Ecklonia cava extract mixture containing Ecklonia cava extract and Chrysanthemum chrysanthemum extract as an active ingredient, and a respiratory product containing an Ecklonia cava extract mixture containing Ecklonia chrysanths extract and Chrysanthemum chrysanthemum extract as an active ingredient. It relates to a pharmaceutical composition for preventing or treating diseases, and a manufacturing method of an Ecklonia cava extract mixture, which has excellent efficacy in preventing, improving, or treating respiratory diseases caused or worsened by increased mucin secretion or accompanied by symptoms of increased mucin secretion. can indicate.
Description
감태추출물혼합물을 유효성분으로 포함하는 호흡기 질환 예방, 개선 또는 치료용 조성물 및 제조방법에 관한 것으로, 보다 상세하게는 감태추출물 및 감국추출물을 함유하는 감태추출물혼합물을 유효성분으로 포함하는 호흡기 질환 예방 또는 개선용 건강기능식품 조성물, 및 감태추출물 및 감국추출물을 함유하는 감태추출물혼합물을 유효성분으로 포함하는 호흡기 질환 예방 또는 치료용 약학적 조성물, 및 감태추출물혼합물의 제조방법에 관한 것이다.It relates to a composition and manufacturing method for preventing, improving or treating respiratory diseases containing an Ecklonia cava extract mixture as an active ingredient, and more specifically, to a composition for preventing or manufacturing respiratory diseases containing an Ecklonia cava extract mixture containing Ecklonia cava extract and Chrysanthemum chrysanthemum extract as an active ingredient. It relates to a health functional food composition for improvement, a pharmaceutical composition for preventing or treating respiratory diseases containing an Ecklonia cava extract mixture containing Ecklonia cava extract and Chrysanthemum chrysanthemum extract as an active ingredient, and a method for producing the Ecklonia cava extract mixture.
기도에서의 점액 생성은 폐 건강을 유지하기 위한 중요한 방어기작이다. 그러나 지나친 점액분비는 기도폐쇄 내지 기도과민증을 유발할 수 있으며, 기관지염(bronchitis), 천식(asthma), 만성 폐색성 폐질환(Chronic obstructive pulmonary disease; COPD), 폐렴(pneumonia) 등에서 점액의 과다분비는 병리학적 특징이다.Mucus production in the airways is an important defense mechanism to maintain lung health. However, excessive mucus secretion can cause airway obstruction or airway hyperresponsiveness, and excessive mucus secretion in bronchitis, asthma, chronic obstructive pulmonary disease (COPD), pneumonia, etc. is a pathology. It is a characteristic feature.
뮤신(mucin)은 점액에 점탄성(viscoelasticity)을 부여하는 당단백질로서 뮤신 그룹 유전자(MUS)로 암호화된다. 지금까지 알려진 18가지의 인간 MUS 중, 기도(airway)에서 발견되는 것은 MUC2, MUC4, MUC5AC 및 MUC5B 등으로, 그 중 MUC5AC는 주로 기도상피세포 내지 점액분비세포(airway goblet cell)에서 발현된다. MUC5AC는 기관지 질환 환자의 증상이 악화될수록 분비가 증가하며, 비정상적인 점액 내지 점액세포 증식의 원인인 것으로 알려져 있다. 다만, 뮤신의 과잉생산과 점액세포 증식은 서로 관련이 있지만 동의어는 아니며, 서로 다른 신호 내지 유전자 조절 경로를 따를 수 있다.Mucin is a glycoprotein that provides viscoelasticity to mucus and is encoded by the mucin group gene (MUS). Among the 18 types of human MUS known so far, those found in the airway include MUC2, MUC4, MUC5AC, and MUC5B. Among them, MUC5AC is mainly expressed in airway epithelial cells and mucus secreting cells (airway goblet cells). MUC5AC secretion increases as the symptoms of patients with bronchial disease worsen, and it is known to be the cause of abnormal mucus or mucus cell proliferation. However, although mucin overproduction and mucous cell proliferation are related, they are not synonymous and may follow different signaling or gene regulation pathways.
흡연자 또는 노인층에서 많이 발병하는 기관지 염증은 삶의 질을 저하시킬 뿐만 아니라 이차적인 감염질환의 위험성을 증가시킨다. 또한 노인들은 체력이 떨어져 점액(객담)의 배출이 어려운 경우가 많아 만성 폐색성 폐질환으로까지 진행될 우려가 있어 반드시 초기에 개선시켜야 한다.Bronchial inflammation, which occurs frequently in smokers or the elderly, not only reduces quality of life but also increases the risk of secondary infectious diseases. In addition, elderly people often have difficulty expelling mucus (sputum) due to low physical strength, so there is a risk that it may progress to chronic obstructive pulmonary disease, so it must be improved at an early stage.
감태추출물에 관한 종래기술로는 체중 감소용 감태추출물 및 이의 제조방법(대한민국 공개특허공보 제10-2014-0136326호), 감태추출물을 유효성분으로 포함하는 통증의 완화, 예방 또는 치료용 조성물(대한민국 공개특허공보 제10-2015-0145313호), 감태추출물을 유효성분으로 함유하는 노인성 근간소증 예방 또는 치료용 조성물(대한민국 공개특허공보 제10-2022-0157231호), 갈조류로부터 플로로탄닌 정제물의 제조방법(대한민국 등록특허공보 제10-1592251호) 등이 개시되어 있다.Conventional technologies related to Ecklonia cava extract include Ecklonia cava extract for weight loss and its manufacturing method (Korea Patent Publication No. 10-2014-0136326), and a composition for alleviating, preventing or treating pain containing Ecklonia cava extract as an active ingredient (Republic of Korea) Publication of Patent No. 10-2015-0145313), Composition for preventing or treating senile myoclonus containing Ecklonia cava extract as an active ingredient (Korean Patent Publication No. 10-2022-0157231), Preparation of purified phlorotannins from brown algae A method (Korean Patent Publication No. 10-1592251) is disclosed.
또한, 감국추출물에 관한 종래기술로는 감국추출물을 함유하는 당뇨병, 당뇨합병증의 예방 및 치료용 약학조성물(대한민국 공개특허공보 제10-2009-0106700호), 감국추출물 또는 분획물을 유효성분으로 포함하는 비만, 비만 관련 질환 또는 합병증의 예방, 개선 또는 치료용 조성물(대한민국 공개특허공보 제10-2017-0088314호), 흰색 감국추출물을 포함하는 뇌하혈 질환 예방 또는 개선용 조성물(대한민국 공개특허공보 제10-2009-0076195호) 등이 개시되어 있다. In addition, the prior art regarding chrysanthemum extract includes a pharmaceutical composition for preventing and treating diabetes and diabetic complications containing chrysanthemum extract (Korean Patent Publication No. 10-2009-0106700), and a pharmaceutical composition containing chrysanthemum chrysanthemum extract or fractions as an active ingredient. Composition for preventing, improving or treating obesity, obesity-related diseases or complications (Korean Patent Publication No. 10-2017-0088314), Composition for preventing or improving subcerebrovascular disease containing white chrysanthemum extract (Korean Patent Publication No. 10) -2009-0076195) etc. are disclosed.
그러나, 감태추출물혼합물을 유효성분으로 포함하는 호흡기 질환 예방, 개선 또는 치료용 조성물에 대해서는 알려져 있지 아니한 실정이다. 이에 본 발명자들은 천연 자원인 식물 추출물의 복합물을 사용하여 과도한 점액 분비를 조절하고 염증성 호흡기 질환을 다스리기 위한 방안을 연구하여 본 발명을 완성하였다.However, there is no known composition for preventing, improving or treating respiratory diseases containing the Ecklonia cava extract mixture as an active ingredient. Accordingly, the present inventors studied a method for controlling excessive mucus secretion and treating inflammatory respiratory diseases using a complex of plant extracts, which are natural resources, and completed the present invention.
본 발명의 목적은 뮤신분비의 증가에 의해 발생 또는 악화되거나 뮤신분비가 증가하는 증상을 동반하는 호흡기질환에 유효한 감태추출물혼합물을 포함하는 호흡기 질환 예방, 개선 또는 치료용 조성물을 제공하는 것이다.The purpose of the present invention is to provide a composition for preventing, improving or treating respiratory diseases containing an Ecklonia cava extract mixture that is effective for respiratory diseases caused or worsened by increased mucin secretion or accompanied by symptoms of increased mucin secretion.
본 발명의 다른 목적은 뮤신분비의 증가에 의해 발생 또는 악화되거나 뮤신분비가 증가하는 증상을 동반하는 호흡기질환에 유효한 감태추출물혼합물의 제조방법을 제공하는 것이다.Another object of the present invention is to provide a method for producing an Ecklonia cava extract mixture that is effective for respiratory diseases caused or worsened by increased mucin secretion or accompanied by symptoms of increased mucin secretion.
본 발명은 상기와 같은 목적을 달성하기 위하여, 감태추출물 및 감국추출물을 함유하는 감태추출물혼합물을 유효성분으로 포함하는 호흡기 질환 예방 또는 개선용 건강기능식품 조성물을 제공한다.In order to achieve the above object, the present invention provides a health functional food composition for preventing or improving respiratory diseases containing an Ecklonia cava extract and an Ecklonia chrysanthemum extract mixture containing Chrysanthemum chrysanthemum extract as an active ingredient.
상기 호흡기 질환 예방 또는 개선용 건강기능식품 조성물에서, 상기 감태추출물혼합물은 상기 감태추출물 및 상기 감국추출물을 10~90 : 10~90의 중량비로 함유할 수 있다.In the health functional food composition for preventing or improving respiratory diseases, the Ecklonia cava extract mixture may contain the Ecklonia cava extract and the Chrysanthemum chrysanthemum extract at a weight ratio of 10 to 90:10 to 90.
상기 호흡기 질환 예방 또는 개선용 건강기능식품 조성물에서, 상기 감태추출물혼합물은 상기 감태추출물 및 상기 감국추출물을 50 : 50의 중량비로 함유할 수 있다.In the health functional food composition for preventing or improving respiratory diseases, the Ecklonia cava extract mixture may contain the Ecklonia cava extract and the Chrysanthemum chrysanthemum extract at a weight ratio of 50:50.
상기 호흡기 질환 예방 또는 개선용 건강기능식품 조성물에서, 상기 감태추출물혼합물의 농도는 90~110 ㎍/mL일 수 있다.In the health functional food composition for preventing or improving respiratory diseases, the concentration of the Ecklonia cava extract mixture may be 90 to 110 μg/mL.
상기 호흡기 질환 예방 또는 개선용 건강기능식품 조성물에서, 상기 호흡기 질환은 만성폐쇄성폐질환(COPD), 천식, 폐암, 하기도 및 호흡기계 증상 감기, 인플루엔자, 인두염, 기침, 가래, 기관지염, 폐기능 감소 및 폐렴으로 이루어진 군에서 선택되는 하나 이상일 수 있다.In the health functional food composition for preventing or improving respiratory diseases, the respiratory diseases include chronic obstructive pulmonary disease (COPD), asthma, lung cancer, lower respiratory tract and respiratory system symptoms cold, influenza, pharyngitis, cough, phlegm, bronchitis, decreased lung function, and It may be one or more selected from the group consisting of pneumonia.
상기 호흡기 질환 예방 또는 개선용 건강기능식품 조성물에서, 상기 감태추출물혼합물의 농도 100 ㎍/mL에서, 100 ㎍/mL 농도의 감태추출물에 비하여, 염증성 점액단백질 생성 억제능이 90~110% 증가하고, 100 ㎍/mL 농도의 감국추출물에 비하여, 염증성 점액단백질 생성 억제능이 45~55% 증가할 수 있다.In the health functional food composition for preventing or improving respiratory diseases, at a concentration of 100 μg/mL of the Ecklonia cava extract mixture, the ability to inhibit inflammatory mucus protein production increases by 90 to 110% compared to the Ecklonia cava extract at a concentration of 100 μg/mL, and 100 Compared to chrysanthemum extract at a ㎍/mL concentration, the ability to inhibit inflammatory mucus protein production can increase by 45-55%.
본 발명은 상기와 같은 목적을 달성하기 위하여, 감태추출물 및 감국추출물을 함유하는 감태추출물혼합물을 유효성분으로 포함하는 호흡기 질환 예방 또는 치료용 약학적 조성물을 제공한다.In order to achieve the above object, the present invention provides a pharmaceutical composition for preventing or treating respiratory diseases containing an Ecklonia cava extract and an Ecklonia cava extract mixture containing Chrysanthemum chrysanthemum extract as an active ingredient.
상기 호흡기 질환 예방 또는 치료용 약학적 조성물에서, 상기 감태추출물혼합물은 상기 감태추출물 및 상기 감국추출물을 10~90 : 10~90의 중량비로 함유할 수 있다.In the pharmaceutical composition for preventing or treating respiratory diseases, the Ecklonia cava extract mixture may contain the Ecklonia cava extract and the Chrysanthemum chrysanthemum extract at a weight ratio of 10 to 90:10 to 90.
상기 호흡기 질환 예방 또는 치료용 약학적 조성물에서, 상기 감태추출물혼합물은 상기 감태추출물 및 상기 감국추출물을 50 : 50의 중량비로 함유할 수 있다.In the pharmaceutical composition for preventing or treating respiratory diseases, the Ecklonia cava extract mixture may contain the Ecklonia cava extract and the Chrysanthemum chrysanthemum extract at a weight ratio of 50:50.
상기 호흡기 질환 예방 또는 치료용 약학적 조성물에서, 상기 감태추출물혼합물의 농도는 90~110 ㎍/mL일 수 있다.In the pharmaceutical composition for preventing or treating respiratory diseases, the concentration of the Ecklonia cava extract mixture may be 90 to 110 μg/mL.
상기 호흡기 질환 예방 또는 치료용 약학적 조성물에서, 상기 호흡기 질환은 만성폐쇄성폐질환(COPD), 천식, 폐암, 하기도 및 호흡기계 증상 감기, 인플루엔자, 인두염, 기침, 가래, 기관지염, 폐기능 감소 및 폐렴으로 이루어진 군에서 선택되는 하나 이상일 수 있다.In the pharmaceutical composition for preventing or treating respiratory diseases, the respiratory diseases include chronic obstructive pulmonary disease (COPD), asthma, lung cancer, lower respiratory tract and respiratory system symptoms, cold, influenza, pharyngitis, cough, phlegm, bronchitis, decreased lung function, and pneumonia. It may be one or more selected from the group consisting of.
상기 호흡기 질환 예방 또는 치료용 약학적 조성물에서, 상기 감태추출물혼합물의 농도 100 ㎍/mL에서, 100 ㎍/mL 농도의 감태추출물에 비하여, 염증성 점액단백질 생성 억제능이 90~110% 증가하고, 100 ㎍/mL 농도의 감국추출물에 비하여, 염증성 점액단백질 생성 억제능이 45~55% 증가할 수 있다.In the pharmaceutical composition for preventing or treating respiratory diseases, at a concentration of 100 μg/mL of the Ecklonia cava extract mixture, the ability to inhibit inflammatory mucus protein production increases by 90-110% compared to the Ecklonia cava extract at a concentration of 100 μg/mL, and 100 μg Compared to Chrysanthemum chrysanthemum extract at a /mL concentration, the ability to inhibit inflammatory mucus protein production can increase by 45-55%.
본 발명은 상기와 같은 목적을 달성하기 위하여, (a) 감태를 세척한 후 추출한 다음, 여과하여 농축시킨 후 정제한 다음, 농축시킨 후 건조하여 감태추출물을 제조하는 단계; (b) 감국을 세척한 후 추출한 다음, 여과하여 농축시킨 후 부형제를 혼합한 다음, 건조하여 감국추출물을 제조하는 단계; 및 (c) 상기 (a) 단계에서 제조한 감태추출물 및 상기 (b) 단계에서 제조한 감국추출물을 혼합하는 단계를 포함하는 감태추출물혼합물의 제조방법을 제공한다. In order to achieve the above object, the present invention includes the steps of (a) washing and extracting Ecklonia cava, filtering and concentrating, purifying, concentrating and drying to produce Ecklonia cava extract; (b) washing and extracting chrysanthemum chrysanthemum, filtering and concentrating, mixing excipients, and then drying to prepare chrysanthemum chrysanthemum extract; and (c) mixing the Ecklonia cava extract prepared in step (a) and the chrysanthemum chrysanthemum extract prepared in step (b).
본 발명에 따른 상기 호흡기 질환 예방 또는 개선용 건강기능식품 조성물 및 호흡기 질환 예방 또는 치료용 약학적 조성물은 뮤신 분비를 효과적으로 감소시키는 활성을 갖기 때문에, 뮤신 분비의 증가에 의해 발생 또는 악화되거나 뮤신 분비가 증가하는 증상을 동반하는 호흡기질환의 예방, 개선 또는 치료에 우수한 효능을 나타낼 수 있다.Since the health functional food composition for preventing or improving respiratory diseases and the pharmaceutical composition for preventing or treating respiratory diseases according to the present invention have the activity of effectively reducing mucin secretion, it may be caused or worsened by an increase in mucin secretion or mucin secretion. It can show excellent efficacy in preventing, improving, or treating respiratory diseases accompanied by increasing symptoms.
또한, 천연 자원에서 유래한 식물 추출물인 감태추출물 및 감국추출물을 사용하기 때문에 별다른 부작용 없이 호흡기질환의 환자에게 적용할 수 있는 이점이 있다.In addition, because it uses plant extracts derived from natural resources, such as Ecklonia cava extract and Chrysanthemum chrysanthemum extract, it has the advantage of being applicable to patients with respiratory diseases without any side effects.
도 1은 본 발명의 일 실시예에 따른 감태추출물혼합물의 제조공정도를 나타낸 것이다.
도 2는 본 발명의 일 실시예에 따른 실험예 1에서, 감태추출물혼합물의 세포생존율 측정 결과를 나타낸 것이다.
도 3은 본 발명의 일 실시예에 따른 실험예 1에서, 감태추출물혼합물의 NO 생성 저해 효과를 나타낸 것이다.
도 4는 본 발명의 일 실시예에 따른 실험예 2에서, 감태추출물혼합물의 세포생존율 측정 결과를 나타낸 것이다.
도 5는 본 발명의 일 실시예에 따른 실험예 3에서, 인간 유래 기관지 상피세포에서 감테감국 복합물의 염증성 점액단백질 생성 억제 효과를 나타낸 것이다. Figure 1 shows a manufacturing process diagram of an Ecklonia cava extract mixture according to an embodiment of the present invention.
Figure 2 shows the results of measuring cell viability of the Ecklonia cava extract mixture in Experimental Example 1 according to an embodiment of the present invention.
Figure 3 shows the NO production inhibition effect of the Ecklonia cava extract mixture in Experimental Example 1 according to an embodiment of the present invention.
Figure 4 shows the results of measuring cell viability of the Ecklonia cava extract mixture in Experimental Example 2 according to an embodiment of the present invention.
Figure 5 shows the inhibitory effect of the gamtegamguk complex on inflammatory mucus protein production in human-derived bronchial epithelial cells in Experimental Example 3 according to an embodiment of the present invention.
이하, 본 발명을 보다 상세히 설명한다. Hereinafter, the present invention will be described in more detail.
본 발명에서, "예방"이라 함은 질환 또는 질병을 보유하고 있다고 진단된 적은 없으나, 이러한 질환 또는 질병에 걸릴 수 있는 개체에서 질환 또는 질병의 발생을 억제하는 것을 의미한다.In the present invention, “prevention” means suppressing the occurrence of a disease or disease in an individual who has not been diagnosed as having the disease or disease but is susceptible to such disease or disease.
본 발명에서, "개선" 또는 "치료"라 함은 개체에서 질환 또는 질병 악화의 억제, 질환 또는 질병의 경감, 또는 질환 또는 질환의 제거를 의미한다.In the present invention, “improvement” or “treatment” means inhibiting disease or worsening of a disease, alleviating a disease or condition, or eliminating a disease or condition in an individual.
본 발명에서, "개체"는 본 발명의 조성물을 투여하여 증상이 호전될 수 있는 질환을 가진 인간을 포함한 동물을 의미한다.In the present invention, “individual” refers to animals, including humans, whose symptoms can be improved by administering the composition of the present invention.
본 발명의 제1 구현예는 감태추출물 및 감국추출물을 함유하는 감태추출물혼합물을 유효성분으로 포함하는 호흡기 질환 예방 또는 개선용 건강기능식품 조성물에 관한 것이다.The first embodiment of the present invention relates to a health functional food composition for preventing or improving respiratory diseases containing an Ecklonia cava extract and an Ecklonia chrysanthemum extract mixture containing Chrysanthemum chrysanthemum extract as an active ingredient.
본 발명의 제2 구현예는 감태추출물 및 감국추출물을 함유하는 감태추출물혼합물을 유효성분으로 포함하는 호흡기 질환 예방 또는 치료용 약학적 조성물에 관한 것이다.The second embodiment of the present invention relates to a pharmaceutical composition for preventing or treating respiratory diseases containing an Ecklonia cava extract and an Ecklonia cava extract mixture containing Chrysanthemum chrysanthemum extract as an active ingredient.
상기 "감태(Ecklonia cava)"는 갈조 식물 다시마목(Laminariales) 미역과의 식물로서 우리나라에는 주로 제주연안 수심 10 m 내외에서 서식하고, 일본에서도 서식하는 것으로 알려져 있고, 길이는 1~2 m이며 줄기는 원기둥 모양이며 밑동은 뿌리 모양을 하고 있다. 감태는 골다공증, 변비, 불면증, 피브 미용, 빈혈, 항암 등의 효과가 있다고 알려져 있다. The " Ecklonia cava " is a plant of the seaweed family of the brown seaweed order Laminariales. It mainly lives in Korea at a depth of about 10 m off the coast of Jeju, and is known to live in Japan as well. It is 1 to 2 m long and has a stem. It is cylindrical in shape and the base is root-shaped. Ecklonia cava is known to be effective in treating osteoporosis, constipation, insomnia, fib beautification, anemia, and anti-cancer.
상기 "감국(Chrysanthemum indicum)"은 한국, 중국, 일본의 산과 들·해안가에서 자라는 국화과의 여러해살이 식물로서, 특유한 향기가 있고 약성은 조금 달며 쓰고 차다. 눈과 머리를 시원하게 하고 눈물이 나는 것을 멎게 하며 열을 내려 주고, 가슴속에 열이 있어 답답한 증상, 폐렴, 기관지염, 두통, 어깨결림, 고혈압에 효과가 있다고 알려져 있으며, 간기능과 충혈, 생리불순과 여드름 등과 같은 피부 트러블 등에 사용한다.The " Chrysanthemum indicum " is a perennial plant of the Asteraceae family that grows in the mountains, fields, and coastal areas of Korea, China, and Japan. It has a unique scent and a slightly sweet, bitter, and cold taste. It is known to cool the eyes and head, stop tears, and reduce fever. It is also known to be effective in treating symptoms of stuffiness due to heat in the chest, pneumonia, bronchitis, headaches, stiff shoulders, and high blood pressure. It is also known to be effective in treating liver function, congestion, menstrual irregularities, and other symptoms. It is used for skin problems such as acne.
상기 감태 및 감국은 식물 전체 또는 지상부 또는 지하부일 수 있다.The Ecklonia cava and Chrysanthemum chrysanthemum may be the whole plant or the above-ground or underground part.
본 발명에서, 상기 감태추출물 및 감국추출물은 통상적으로 알려진 식물 추출물의 추출 방법을 제한없이 사용하여 제조될 수 있다. In the present invention, the Ecklonia cava extract and Chrysanthemum chrysanthemum extract can be prepared using commonly known plant extract extraction methods without limitation.
본 발명에서, 상기 감태추출물 및 감국추출물의 추출 방법으로는 여과법, 용매 추출, 침지 추출, 환류냉각 추출, 가압추출, 아임계추출, 초임계추출 및 초음파추출 등 당업계의 통상적인 방법을 이용할 수 있으나, 이에 제한되는 것은 아니다.In the present invention, conventional methods in the art such as filtration, solvent extraction, immersion extraction, reflux cooling extraction, pressure extraction, subcritical extraction, supercritical extraction and ultrasonic extraction can be used as extraction methods for the Ecklonia cava extract and chrysanthemum chrysanthemum extract. However, it is not limited to this.
상기 감태추출물 및 감국추출물의 추출 방법으로는 바람직하게는 용매 추출일 수 있다. 상기 용매 추출의 용매로는 탄소수 1-4의 저급 알코올(예: 메탄올, 에탄올, 프로판올, 부탄올), 주정, 헥산, 물 또는 이들의 혼합물일 수 있으나, 이에 제한되는 것은 아니다. The extraction method for the Ecklonia cava extract and Chrysanthemum chrysanthemum extract may preferably be solvent extraction. The solvent for the solvent extraction may be a lower alcohol having 1 to 4 carbon atoms (e.g., methanol, ethanol, propanol, butanol), alcohol, hexane, water, or a mixture thereof, but is not limited thereto.
주정을 추출 용매로 사용하는 경우, 주정의 농도는 20~80%(v/v)일 수 있으나, 이에 제한되는 것은 아니다.When alcohol is used as an extraction solvent, the concentration of alcohol may be 20 to 80% (v/v), but is not limited thereto.
상기 용매는 상기 감태 또는 감국의 식물체 중량에 대하여 1~15배, 바람직하게는 2~10배, 보다 바람직하게는 3~7배일 수 있으나, 이에 제한되는 것은 아니다.The solvent may be 1 to 15 times, preferably 2 to 10 times, and more preferably 3 to 7 times the weight of the Ecklonia cava or Chrysanthemum chrysanthemum plant, but is not limited thereto.
본 발명의 상기 건강기능식품 조성물 및 약학적 조성물은 뮤신분비의 증가에 의해 발생 또는 악화되거나 뮤신 분비가 증가하는 증상을 동반하는 호흡기질환의 예방, 개선 또는 치료를 위한 조성물로서, 뮤신 분비를 감소시키는 활성을 갖는다.The health functional food composition and pharmaceutical composition of the present invention are compositions for preventing, improving or treating respiratory diseases caused or worsened by increased mucin secretion or accompanied by symptoms of increased mucin secretion, and are used to reduce mucin secretion. It has activity.
상기 호흡기 질환은 만성폐쇄성폐질환(COPD), 천식, 폐암, 하기도 및 호흡기계 증상 감기, 인플루엔자, 인두염, 기침, 가래, 기관지염, 폐기능 감소 및 폐렴으로 이루어진 군에서 선택되는 하나 이상일 수 있으나, 이에 제한되는 것은 아니며 뮤신 또는 객담이 과다 분비되어 발생 또는 악화되거나 이러한 증상을 동반하는 질환이면 그 종류에 상관없이 적용될 수 있다.The respiratory disease may be one or more selected from the group consisting of chronic obstructive pulmonary disease (COPD), asthma, lung cancer, lower respiratory tract and respiratory system symptoms, cold, influenza, pharyngitis, cough, phlegm, bronchitis, decreased lung function, and pneumonia. It is not limited and can be applied regardless of the type of disease as long as it is caused or worsened by excessive secretion of mucin or sputum or is accompanied by these symptoms.
상기 뮤신은 MUC2, MUC4, MUC5AC 및 MUC5B 중 하나 이상일 수 있고, 구체적으로는 MUC5AC일 수 있으나, 이에 제한되는 것은 아니다.The mucin may be one or more of MUC2, MUC4, MUC5AC, and MUC5B, and may specifically be MUC5AC, but is not limited thereto.
본 발명의 상기 건강기능식품 조성물 및 약학적 조성물에서, 상기 감태추출물혼합물은 상기 감태추출물 및 상기 감국추출물을 10~90 : 10~90의 중량비로 함유할 수 있다. 예를 들면, 80:20의 중량비, 60:40의 중량비, 50:50의 중량비, 40:60의 중량비, 20:80의 중량비로 함유될 수 있으나, 이에 제한되는 것은 아니다.In the health functional food composition and pharmaceutical composition of the present invention, the Ecklonia cava extract mixture may contain the Ecklonia cava extract and the Chrysanthemum chrysanthemum extract at a weight ratio of 10 to 90:10 to 90. For example, it may be contained in a weight ratio of 80:20, 60:40, 50:50, 40:60, or 20:80, but is not limited thereto.
본 발명의 상기 건강기능식품 조성물 및 약학적 조성물에서, 상기 감태추출물혼합물의 농도는 90~110 ㎍/mL, 바람직하게는 95~105 ㎍/mL, 보다 바람직하게는 105 ㎍/mL일 수 있으나, 이에 제한되는 것은 아니다.In the health functional food composition and pharmaceutical composition of the present invention, the concentration of the Ecklonia cava extract mixture may be 90 to 110 μg/mL, preferably 95 to 105 μg/mL, and more preferably 105 μg/mL. It is not limited to this.
본 발명의 상기 건강기능식품 조성물 및 약학적 조성물에서, 상기 감태추출물혼합물의 농도 100 ㎍/mL에서, 100 ㎍/mL 농도의 감태추출물에 비하여 염증성 점액단백질 생성 억제능이 90~110% 증가하고, 100 ㎍/mL 농도의 감국추출물에 비하여 염증성 점액단백질 생성 억제능이 45~55% 증가할 수 있으나, 이에 제한되는 것은 아니다.In the health functional food composition and pharmaceutical composition of the present invention, at a concentration of 100 μg/mL of the Ecklonia cava extract mixture, the ability to inhibit inflammatory mucus protein production increases by 90 to 110% compared to the Ecklonia cava extract at a concentration of 100 μg/mL, and 100 μg/mL. Compared to chrysanthemum extract at a concentration of ㎍/mL, the ability to inhibit inflammatory mucus protein production may increase by 45-55%, but is not limited to this.
본 발명의 제1 구현예에 따른 상기 건강기능식품 조성물에서, 상기 "건강기능식품"은 인체에 유용한 기능성을 가진 원료나 성분을 사용하여 정제, 캅셀, 분말, 과립, 액상 및 환 등의 형태로 제조 및 가공한 식품을 말한다. 여기서 “기능성”이라 함은 인체의 구조 및 기능에 대하여 영양소를 조절하거나 생리학적 작용 등과 같은 보건용도에 유용한 효과를 얻는 것을 의미한다.In the health functional food composition according to the first embodiment of the present invention, the “health functional food” is in the form of tablets, capsules, powders, granules, liquids, and pills using raw materials or ingredients with functional properties useful to the human body. Refers to manufactured and processed food. Here, “functionality” means controlling nutrients for the structure and function of the human body or obtaining useful effects for health purposes, such as physiological effects.
본 발명의 상기 건강기능식품은 통상의 기술분야에서 통상적으로 사용되는 방법에 의해 제조 가능하며, 상기 제조시에는 통상의 기술분야에서 통상적으로 첨가하는 원료 및 성분을 첨가하여 제조할 수 있다. 또한, 상기 건강기능식품의 제형 또한 건강기능식품으로 인정되는 제형이면 제한없이 제조될 수 있다.The health functional food of the present invention can be manufactured by a method commonly used in the field of technology, and can be manufactured by adding raw materials and components commonly added in the field of technology. Additionally, the formulation of the health functional food can also be manufactured without limitation as long as it is a formulation recognized as a health functional food.
본 발명의 건강기능식품 조성물은 다양한 형태의 제형으로 제조될 수 있으며, 일반 약품과는 달리 식품을 원료로 하여 약품의 장기 복용시 발생할 수 있는 부작용 등이 없는 장점이 있고, 휴대성이 뛰어나, 본 발명의 건강기능식품은 항산화제의 효과를 증진시키기 위한 보조제로 섭취가 가능하다.The health functional food composition of the present invention can be manufactured in various forms, and unlike general drugs, it is made from food as a raw material and has the advantage of having no side effects that may occur when taking the drug for a long time, and is excellent in portability. The health functional food of the invention can be consumed as a supplement to enhance the effect of antioxidants.
본 발명의 상기 건강기능식품 조성물은 유효성분 이외에 식품학적으로 허용가능한 식품보조첨가제를 포함할 수 있으며, 유효성분의 혼합량은 사용 목적(예방, 건강 또는 치료적 처치)에 따라 적합하게 결정될 수 있다.The health functional food composition of the present invention may include food additives that are foodologically acceptable in addition to the active ingredients, and the mixing amount of the active ingredients may be appropriately determined depending on the purpose of use (prevention, health, or therapeutic treatment).
본 발명에서, 상기 "식품보조첨가제"라 함은 식품에 보조적으로 첨가될 수 있는 구성요소를 의미하며, 각 제형의 건강기능식품 또는 건강식품을 제조하는데 첨가되는 것으로서 당업자가 적절히 선택하여 사용할 수 있다. 식품보조첨가제의 예로는 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, 안정 화제, 방부제, 글리세린, 알콜, 탄산음료에 사용되는 탄산화제 등이 포함되지만, 상기 예들에 의해 본 발명의 식품보조첨가제의 종류가 제한되는 것은 아니다.In the present invention, the term "food supplement" refers to a component that can be added to food as an auxiliary agent, and can be appropriately selected and used by a person skilled in the art as it is added to manufacture each type of health functional food or health food. . Examples of food supplements include various nutrients, vitamins, minerals (electrolytes), flavoring agents such as synthetic and natural flavors, colorants and fillers, pectic acid and its salts, alginic acid and its salts, organic acids, and protective colloidal thickeners. , pH adjusters, stabilizers, preservatives, glycerin, alcohol, carbonating agents used in carbonated beverages, etc., but the types of food supplements of the present invention are not limited to the above examples.
본 발명의 제2 구현예에 따른 상기 약학적 조성물은 당업계에서 약학적 조성물의 제조에 통상적으로 사용하는 적절한 담체, 부형제 또는 희석제를 추가로 포함할 수 있다. 바람직하게 상기 약학적 조성물은 정제, 환제, 산제, 과립제, 캡슐제, 현탁제, 내용액제, 유제, 시럽제, 멸균된 수용액, 비수성용제, 현탁제, 유제, 동결건조제제, 경피흡수제, 겔제, 로션제, 연고제, 크림제, 첩부제, 카타플라스마제, 페이스트제, 스프레이, 피부 유화액, 피부 현탁액, 경피 전달성 패치, 약물 함유 붕대 및 좌제로 이루어진 군으로부터 선택되는 어느 하나의 제형을 가질 수 있으며, 경구 또는 비경구의 여러 가지 제형일 수 있으나, 이에 제한되는 것은 아니다.The pharmaceutical composition according to the second embodiment of the present invention may further include an appropriate carrier, excipient, or diluent commonly used in the preparation of pharmaceutical compositions in the art. Preferably, the pharmaceutical composition is tablets, pills, powders, granules, capsules, suspensions, oral solutions, emulsions, syrups, sterilized aqueous solutions, non-aqueous solvents, suspensions, emulsions, freeze-dried preparations, transdermal absorbents, gels, and lotions. It may have any one dosage form selected from the group consisting of ointments, creams, patches, cataplasms, pastes, sprays, dermal emulsions, dermal suspensions, transdermal delivery patches, drug-containing bandages, and suppositories, It may be in various oral or parenteral dosage forms, but is not limited thereto.
제제화할 경우에는 보통 사용하는 충진제, 증량제, 결합제, 습윤제, 붕해제, 계면활성제 등의 희석제 또는 부형제를 사용하여 조제될 수 있으나, 이에 제한되는 것은 아니다.When formulated, it may be prepared using diluents or excipients such as commonly used fillers, extenders, binders, wetting agents, disintegrants, and surfactants, but is not limited thereto.
경구 투여를 위한 고형 제제에는 정제, 환제, 산제, 과립제, 캡슐제 등이 포함되며, 이러한 고형 제제는 하나 이상의 화합물에 적어도 하나 이상의 부형제 예를 들면, 전분, 탄산칼슘, 수크로오스(sucrose) 또는 락토오스(lactose), 젤라틴 등을 섞어 조제된다. 또한, 단순한 부형제 이외에 스테아린산 마그네슘, 탈크 등과 같은 윤활제들도 사용될 수 있으나, 이에 제한되는 것은 아니다.Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc., and these solid preparations contain one or more compounds and at least one excipient, such as starch, calcium carbonate, sucrose, or lactose ( It is prepared by mixing lactose, gelatin, etc. Additionally, in addition to simple excipients, lubricants such as magnesium stearate, talc, etc. may be used, but are not limited thereto.
경구 투여를 위한 액상 제제로는 현탁제, 내용액제, 유제, 시럽제 등이 해당되는데 흔히 사용되는 단순 희석제인 물, 리퀴드 파라핀 이외에 여러 가지 부형제, 예를 들면 습윤제, 감미제, 방향제, 보존제 등이 포함될 수 있으나, 이에 제한되는 것은 아니다.Liquid preparations for oral administration include suspensions, oral solutions, emulsions, and syrups. In addition to the commonly used simple diluents such as water and liquid paraffin, various excipients such as wetting agents, sweeteners, fragrances, and preservatives may be included. However, it is not limited to this.
비경구 투여를 위한 제제에는 멸균된 수용액, 비수성용제, 현탁제, 유제, 동결건조 제제, 좌제가 포함될 수 있으나, 이에 제한되는 것은 아니다. 비수성 용제, 현탁 용제로는 프로필렌글리콜, 폴리에틸렌 글리콜, 올리브 오일과 같은 식물성 기름, 에틸올레이트와 같은 주사 가능한 에스테르 등이 사용될 수 있으나, 이에 한정되는 되는 것은 아니다. 좌제의 기제로는 위텝솔(witepsol), 마크로골, 트윈 61(tween 61), 카카오지, 라우린지, 글리세로젤라틴 등이 사용될 수 있으나, 이에 제한되는 되는 것은 아니다.Preparations for parenteral administration may include, but are not limited to, sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Non-aqueous solvents and suspension solvents include, but are not limited to, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, and injectable ester such as ethyl oleate. As a base for suppositories, witepsol, macrogol, tween 61, cacao, laurel, glycerogelatin, etc. may be used, but are not limited thereto.
본 발명의 상기 약학적 조성물의 바람직한 투여량은 환자의 상태 및 체중, 질병의 정도, 약물형태, 투여경로 및 기간에 따라 다르지만, 당업자에 의해 적절하게 선택될 수 있다. 그러나, 보다 효과적인 예방 또는 치료를 위하여, 본 발명의 상기 감태추출물혼합물은 1일 0.0001 내지 30 mg/kg으로, 바람직하게는 0.001 내지 10 mg/kg으로 투여 함이 바람직하고, 투여는 하루에 한번 투여할 수도 있고, 수회 나누어 투여할 수도 있다.The preferred dosage of the pharmaceutical composition of the present invention varies depending on the patient's condition and weight, degree of disease, drug form, administration route and period, but can be appropriately selected by a person skilled in the art. However, for more effective prevention or treatment, the Ecklonia cava extract mixture of the present invention is preferably administered at 0.0001 to 30 mg/kg per day, preferably 0.001 to 10 mg/kg, and administered once a day. It may be administered in multiple doses.
본 발명의 약학적 조성물은 쥐, 마우스, 가축, 인간 등의 포유동물에 다양한 경로로 투여될 수 있다. 투여의 모든 방식은 예상될 수 있는데, 예를 들면, 경구, 직장 또는 정맥, 근육, 피하, 자궁 내 경막 또는 뇌혈관 내(intracerebroventricular) 주사에 의해 투여될 수 있다.The pharmaceutical composition of the present invention can be administered to mammals such as rats, mice, livestock, and humans through various routes. All modes of administration are contemplated, for example, oral, rectal or by intravenous, intramuscular, subcutaneous, intrauterine dural or intracerebroventricular injection.
이하, 본 발명의 이해를 돕기 위하여 실시예 등을 들어 상세하게 설명하기로 한다. 그러나, 본 발명에 따른 실시예들은 여러 가지 다른 형태로 변형될 수 있으며, 본 발명의 범위가 하기 실시예들에 한정되는 것으로 해석되어서는 안된다. 본 발명의 실시예들은 당업계에서 평균적인 지식을 가진 자에게 본 발명을 보다 완전하게 설명 하기 위해 제공되는 것이다.Hereinafter, to aid understanding of the present invention, it will be described in detail through examples. However, the embodiments according to the present invention may be modified into various other forms, and the scope of the present invention should not be construed as being limited to the following embodiments. Embodiments of the present invention are provided to more completely explain the present invention to those skilled in the art.
<실시예 1> 감태추출물혼합물의 제조<Example 1> Preparation of Ecklonia cava extract mixture
감태추출물은 원료인 감태(Ecklonia cava)를 추출탱크에 투입하여 세척한 후 70%주정으로 추출하고 추출액을 여과 및 감압 농축하였다. 농축액을 정제한 다음 건조하여 감태추출물을 제조하였다.For the Ecklonia cava extract, the raw material, Ecklonia cava , was put into an extraction tank, washed, extracted with 70% alcohol, and the extract was filtered and concentrated under reduced pressure. The concentrate was purified and dried to prepare Ecklonia cava extract.
감국추출물은 원료인 감국(Chrysanthemum incidicum)을 추출 탱크에 투입하여 세척한 후 50% 주정으로 추출하고 추출액을 여과 및 감압 농축하여 부형제(덱스트린)를 배합후 감국추출물을 제조하였다. Chrysanthemum incidicum , the raw material, was put into an extraction tank, washed, extracted with 50% alcohol, and the extract was filtered and concentrated under reduced pressure to mix with an excipient (dextrin).
상기에서 제조한 감국추출물과 감태추출물을 50:50의 중량비로 혼합하여 감태추출물혼합물을 제조하였다.An Ecklonia cava extract mixture was prepared by mixing the Chrysanthemum chrysanthemum extract and Ecklonia cava extract prepared above in a weight ratio of 50:50.
도 1은 본 발명의 일 실시예에 따른 감태추출물혼합물의 제조공정도를 나타낸 것이다.Figure 1 shows a manufacturing process diagram of an Ecklonia cava extract mixture according to an embodiment of the present invention.
<실시예 2> 감태추출물혼합물의 제조<Example 2> Preparation of Ecklonia cava extract mixture
감국추출물과 감태추출물의 혼합비를 80 : 20의 중량비로 한 것 이외에는 상기 실시예 1과 동일하게 실시하여 감태추출물혼합물을 제조하였다. An Ecklonia cava extract mixture was prepared in the same manner as in Example 1 except that the mixing ratio of Chrysanthemum chrysanthemum extract and Ecklonia cava extract was set to a weight ratio of 80:20.
<실시예 3> 감태추출물혼합물의 제조<Example 3> Preparation of Ecklonia cava extract mixture
감국추출물과 감태추출물의 혼합비를 60 : 40의 중량비로 한 것 이외에는 상기 실시예 1과 동일하게 실시하여 감태추출물혼합물을 제조하였다. An Ecklonia cava extract mixture was prepared in the same manner as in Example 1 except that the mixing ratio of Chrysanthemum chrysanthemum extract and Ecklonia cava extract was set to a weight ratio of 60:40.
<실시예 4> 감태추출물혼합물의 제조<Example 4> Preparation of Ecklonia cava extract mixture
감국추출물과 감태추출물의 혼합비를 40 : 60의 중량비로 한 것 이외에는 상기 실시예 1과 동일하게 실시하여 감태추출물혼합물을 제조하였다. An Ecklonia cava extract mixture was prepared in the same manner as in Example 1 except that the mixing ratio of Chrysanthemum chrysanthemum extract and Ecklonia cava extract was set to a weight ratio of 40:60.
<실시예 5> 감태추출물혼합물의 제조<Example 5> Preparation of Ecklonia cava extract mixture
감국추출물과 감태추출물의 혼합비를 20 : 80의 중량비로 한 것 이외에는 상기 실시예 1과 동일하게 실시하여 감태추출물혼합물을 제조하였다. An Ecklonia cava extract mixture was prepared in the same manner as in Example 1 except that the mixing ratio of Chrysanthemum chrysanthemum extract and Ecklonia cava extract was set to a weight ratio of 20:80.
<실시예 6> 감태추출물혼합물의 제조<Example 6> Preparation of Ecklonia cava extract mixture
감태추출물의 추출 용매를 60% 주정으로 하고, 감국추출물의 추출 용매를 40% 주정으로 한 것 이외에는 상기 실시예 1과 동일하게 실시하여 감태추출물혼합물을 제조하였다. An Ecklonia cava extract mixture was prepared in the same manner as in Example 1 except that the extraction solvent for the Ecklonia cava extract was 60% alcohol and the extraction solvent for the Ecklonia chrysanthemum chrysanthemum extract was 40% alcohol.
<실시예 7> 감태추출물혼합물의 제조<Example 7> Preparation of Ecklonia cava extract mixture
감태추출물의 추출 용매를 50% 주정으로 하고, 감국추출물의 추출 용매를 30% 주정으로 한 것 이외에는 상기 실시예 1과 동일하게 실시하여 감태추출물혼합물을 제조하였다. An Ecklonia cava extract mixture was prepared in the same manner as in Example 1 except that the extraction solvent for the Ecklonia cava extract was 50% alcohol, and the extraction solvent for the Ecklonia chrysanthemum chrysanthemum extract was 30% alcohol.
<실시예 8> 감태추출물혼합물의 제조<Example 8> Preparation of Ecklonia cava extract mixture
감태추출물 및 감국추출물의 추출 용매를 헥산으로 한 것 이외에는 상기 실시예 1과 동일하게 실시하여 감태추출물혼합물을 제조하였다. An Ecklonia cava extract mixture was prepared in the same manner as in Example 1 except that hexane was used as the extraction solvent for the Ecklonia cava extract and Chrysanthemum chrysanthemum extract.
<실험예 1> 항염증 활성 스크리닝<Experimental Example 1> Anti-inflammatory activity screening
상기 실시예 1에서 제조한 감태추출물혼합물을 대상으로 하기와 같이 항염증 활성을 스크리닝하였다.The Ecklonia cava extract mixture prepared in Example 1 was screened for anti-inflammatory activity as follows.
1. 세포주 배양1. Cell line culture
본 실험에 사용된 RAW 264.7 (mouse macrophage cell line) 세포는 한국 세포주은행에서 분양받아 사용하였다. RAW264.7 세포는 각각 10% 소태아 혈청(fetal bovine serum; FBS), 1% 페니실린(P/S)을 첨가한 DMEM 배지에서 5% CO2가 존재하는 37℃ 배양기에서 2~3일에 한 번씩 계대 배양해 주었다.RAW 264.7 (mouse macrophage cell line) cells used in this experiment were purchased from the Korean Cell Line Bank. RAW264.7 cells were cultured in DMEM medium supplemented with 10% fetal bovine serum (FBS) and 1% penicillin (P/S), respectively, in an incubator at 37°C in the presence of 5% CO 2 for 2 to 3 days. They were subcultured every now and then.
2. 세포 생존율 측정2. Measurement of cell viability
RAW264.7 세포에서 감태감국복합물의 세포 생존율을 확인하기 위하여 3-(4,5-디메틸티아졸-2-일)-2,5-디페닐테트라졸리움 브로마이드(MTT)로 세포 생존율(cell viability)를 측정하였다.To confirm the cell viability of Ecklonia Ecklonia complex in RAW264.7 cells, cell viability was measured with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT). was measured.
세포(1x105/well)를 96-웰 플레이트에 200μL씩 분주하여 1시간 동안 CO2 배양기에서 배양 후, 무혈청 배지로 교체 후 시료를 각 농도별로 처리하여 24시간 뒤에 10% MTT를 100μL씩 분주하여 1시간 동안 반응시켜 MTT가 환원되도록 하였다. Cells (1x10 5 /well) were dispensed at 200 μL each into a 96-well plate and cultured in a CO 2 incubator for 1 hour. Then, replaced with serum-free medium, samples were treated at each concentration, and 24 hours later, 10% MTT was dispensed at 100 μL each. The reaction was performed for 1 hour to reduce MTT.
그 후 상등액을 제거하고 디메틸설폭사이드(DMSO)를 100μL씩 분주하여 포르마잔(formazan) 된 세포 결정을 용해시켜 마이크로플레이트 스펙트로포토미터를 사용하여 595nm에서 흡광도를 측정하였다. 세포 생존율은 대조군과 비교하여 백분율(%)로 나타내었다.Afterwards, the supernatant was removed, 100 μL of dimethyl sulfoxide (DMSO) was added to dissolve the formazan cell crystals, and the absorbance was measured at 595 nm using a microplate spectrophotometer. Cell viability was expressed as percentage (%) compared to the control group.
리포폴리사카라이드(LPS)는 그램 음성 세균의 세포벽 물질로서 면역 세포를 자극하여 NO의 생성을 증가시킨다고 보고되고 있다. LPS로 산화적 스트레스를 유발한 RAW 264.7 세포에서 감태추출물혼합물을 농도별로 처리하여 일정 시간 배양하였을 때, LPS 자극에 의한 세포독성은 없는 것으로 확인되었다.Lipopolysaccharide (LPS), a cell wall material of Gram-negative bacteria, has been reported to stimulate immune cells and increase NO production. When RAW 264.7 cells induced by oxidative stress with LPS were treated with Ecklonia cava extract mixture at different concentrations and cultured for a certain period of time, it was confirmed that there was no cytotoxicity due to LPS stimulation.
도 2에서 보는 바와 같이, 감태추출물혼합물의 경우 10~1000μg/mL 100% 이상 세포 생존율을 보였다. As shown in Figure 2, the Ecklonia cava extract mixture showed a cell viability of more than 100% at a range of 10 to 1000 μg/mL.
3. NO 생성 저해3. Inhibition of NO production
NO 함량은 Green 등의 방법으로 측정하였다. RAW 264.7 세포를 DMEM 배지를 이용하여 1×105 cells/mL 농도로 96 웰 플레이트에 분주한 후, 시험 시료와 LPS (100 ng/mL, Sigma Co.)를 함유한 새로운 배지를 동시에 처리하여 24시간 배양하였다.NO content was measured by the method of Green et al. RAW 264.7 cells were distributed in a 96-well plate at a concentration of 1 × 10 5 cells/mL using DMEM medium, and then simultaneously treated with new medium containing the test sample and LPS (100 ng/mL, Sigma Co.) for 24 hours. cultured for some time.
세포배양 상등액 100 μL와 Griess 시약 (1% 설파닐아미드, 0.1% 나프틸에틸렌디아미네인, 2.5% 인산) 100 μL를 혼합한 후 ELISA 리더를 이용하여 520nm에서 흡광도를 측정하였으며, 소디움나이트레이트(Sigma Co.)로 표준 곡선을 작성하여 NO의 함량을 산출하였다.After mixing 100 μL of cell culture supernatant and 100 μL of Griess reagent (1% sulfanilamide, 0.1% naphthylethylenediaminene, 2.5% phosphoric acid), absorbance was measured at 520 nm using an ELISA reader, and sodium nitrate ( The content of NO was calculated by creating a standard curve using Sigma Co.).
LPS 자극에 의해 생성된 NO는 정상적인 생리 상태에서 혈관의 항상성, 세포사멸(apoptosis) 유도 작용 등 중요한 생리적인 기능을 매개하지만, 다량의 NO는 정상세포를 죽이고 염증을 유도하여 급성 또는 만성 염증질환의 원인이 되는 물질로 작용하게 된다. 따라서, 효과적인 NO 분비조절은 급성 또는 만성 염증질환의 치료방법으로 알려져 있으며, 이에 대한 연구가 활발히 진행되고 있다.NO produced by LPS stimulation mediates important physiological functions such as vascular homeostasis and apoptosis induction under normal physiological conditions, but large amounts of NO kill normal cells and induce inflammation, causing acute or chronic inflammatory diseases. It acts as a causative agent. Therefore, effective NO secretion control is known as a treatment method for acute or chronic inflammatory diseases, and research on this is actively underway.
LPS(1μg/mL)d에 의해서 유도된 대식세포를 100%로 기준을 잡고 감태추출물혼합물을 10, 30, 100μg/mL의 농도로 처리한 결과, 도 3에서 보는 바와 같이 감태추출물혼합물에 의하여 NO 생성이 농도 의존적으로 감소하는 것을 확인하였다.As a result of treating macrophages induced by LPS (1 μg/mL) at 100% and treating the Ecklonia cava extract mixture at concentrations of 10, 30, and 100 μg/mL, as shown in Figure 3, NO was produced by the Ecklonia cava extract mixture. It was confirmed that production decreased in a concentration-dependent manner.
<실험예 2> 감태추출물혼합물의 뮤신 과분비 억제 활성 평가<Experimental Example 2> Evaluation of mucin hypersecretion inhibition activity of Ecklonia cava extract mixture
상기 실시예 1에서 제조한 감태추출물혼합물을 대상으로 하기와 같이 뮤신 과분비 억제 활성을 평가하였다.The Ecklonia cava extract mixture prepared in Example 1 was evaluated for its mucin hypersecretion inhibitory activity as follows.
1. 세포주 배양1. Cell line culture
본 실험에 사용된 NCI-H292(human pulmonary mucoepidermoid carcinoma cell line) 세포는 한국 세포주 은행에서 분양받아 사용하였다. NCI-H292 세포는 10% FBS, 2mM L-글루타민, 1% 페니실린(P/S), 100μg/mL 스트렙토마이신이 포함된 RPMI 1640 배지를 이용하여 5% CO2가 존재하는 37℃ 배양기에서 2~3일에 한 번씩 계대 배양해 주었다.NCI-H292 (human pulmonary mucoepidermoid carcinoma cell line) cells used in this experiment were purchased from the Korean Cell Line Bank. NCI-H292 cells were grown in RPMI 1640 medium containing 10% FBS, 2mM L-glutamine, 1% penicillin (P/S), and 100 μg/mL streptomycin for 2~2 days in an incubator at 37°C with 5% CO 2 . Subculture was performed once every three days.
2. 세포 생존율 측정2. Measurement of cell viability
NCI-H292 세포에서 감태추출물혼합물의 세포 생존율을 확인하기 위하여, 3-(4,5-디메틸티아졸-2-일)-2,5-디페닐테트라졸리움 브로마이드(MTT)로 세포 생존율을를 측정하였다.To confirm the cell viability of the Ecklonia cava extract mixture in NCI-H292 cells, the cell viability was measured with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT). .
세포 (4x104/well)를 96-웰 플레이트에 200μL씩 분주하여 1시간 동안 CO2 배양기에서 배양 후, 무혈청 배지로 교체 후 시료를 각 농도별로 처리하여 24시간 뒤에 10% MTT를 100μL씩 분주하여 MTT가 환원되도록 하였다.Cells (4x10 4 /well) were distributed at 200 μL each in a 96-well plate and cultured in a CO 2 incubator for 1 hour. Then, the medium was replaced with serum-free medium, and the samples were treated at each concentration. 24 hours later, 10% MTT was dispensed at 100 μL each. This allowed MTT to be reduced.
그 후 상등액을 제거하고 디메틸설폭사이드(DMSO)를 100μL씩 분주하여 포르마잔(formazan) 된 세포 결정을 용해 시켜 마이크로플레이트 스펙트로포토미터를 사용하여 595nm에서 흡광도를 측정하였다. 세포 생존율은 대조군과 비교하여 백분율(%)로 나타내었다.Afterwards, the supernatant was removed, 100 μL of dimethyl sulfoxide (DMSO) was added to dissolve the formazan cell crystals, and the absorbance was measured at 595 nm using a microplate spectrophotometer. Cell viability was expressed as percentage (%) compared to the control group.
phorbol 12-myristate 13-acetate(PMA)로 세포 분화를 유발한 NCI-H292 세포에서 감태추출물혼합물을 농도별로 처리하여 일정 시간 배양하였을 때, 도 4에서 보는 바와 같이 phorbol 12-myristate 13-acetate(PMA) 자극에 의한 세포독성은 없는 것으로 확인되었다.When NCI-H292 cells, which had induced cell differentiation with phorbol 12-myristate 13-acetate (PMA), were treated with Ecklonia cava extract mixtures at different concentrations and cultured for a certain period of time, as shown in Figure 4, phorbol 12-myristate 13-acetate (PMA) ) It was confirmed that there was no cytotoxicity due to stimulation.
<실험예 3> 인간 유래 기관지 상피세포에서 감국추출물의 염증성 점액단백질 생성 억제 확인<Experimental Example 3> Confirmation of inhibition of inflammatory mucus protein production by chrysanthemum extract in human-derived bronchial epithelial cells
상기 실시예 1에서 제조한 감태추출물혼합물을 대상으로 하기와 같이 인간 유래 기관지 상피세포에서 감국추출물의 염증성 점액단백질 생성 억제 확인하였다.The inhibition of inflammatory mucus protein production by the Ecklonia chrysanthemum chrysanthemum extract in human-derived bronchial epithelial cells was confirmed as follows using the Ecklonia cava extract mixture prepared in Example 1 above.
1. 세포주 배양1. Cell line culture
본 실험에 사용된 NCI-H292(human pulmonary mucoepidermoid carcinoma cell line) 세포는 한국 세포주은행에서 분양 받아 사용하였다. NCI-H292 세포는 10% FBS, 2 mM L-글루타민, 1% 페니실린(P/S), 100μg/mL 스트렙토마이신이 포함된 RPMI 1640 배지를 이용하여 5% CO2가 존재하는 37℃ 배양기에서 2~3일에 한 번씩 계대 배양해 주었다.NCI-H292 (human pulmonary mucoepidermoid carcinoma cell line) cells used in this experiment were purchased from the Korean Cell Line Bank. NCI-H292 cells were grown in an incubator at 37°C in the presence of 5% CO 2 using RPMI 1640 medium containing 10% FBS, 2 mM L-glutamine, 1% penicillin (P/S), and 100 μg/mL streptomycin. Subcultures were performed every ~3 days.
2. 세포 생존율 측정2. Measurement of cell viability
NCI-H292 세포에서 감태추출물혼합물의 MUC5AC 억제 활성을 알아보기 위하여, 인간 유래 기관지 상피세포주 NCI-H292 세포를 4×10⁴cell/mL로 24-웰 플레이트에 500μL씩 분주하고 37℃, 5% CO2 배양기에서 48시간 동안 배양 배지로 배양하여 세포를 안정화하였다.To examine the MUC5AC inhibitory activity of the Ecklonia cava extract mixture in NCI-H292 cells, 500 μL of human-derived bronchial epithelial cell line NCI-H292 cells were dispensed at 4 The cells were stabilized by culturing with culture medium for 48 hours.
48시간 뒤 배양 배지를 모두 제거하고 0.2% FBS/DMEM 배지로 교체한 후 24시간 동안 배양하였다. 그 다음, PBS로 2번 세척하고, 무혈청 배지 기관지 상피세포에 감태, 감국, 감태추출물혼합물을 각각 50μL씩 처리하고 1시간 동안 배양하였다.After 48 hours, all culture medium was removed, replaced with 0.2% FBS/DMEM medium, and cultured for 24 hours. Next, the cells were washed twice with PBS, and the bronchial epithelial cells in serum-free medium were treated with 50 μL each of Ecklonia cava, Chrysanthemum chrysanthemum, and Ecklonia cava extract mixtures and cultured for 1 hour.
그 후 100ng/mL 농도의 PMA를 처리하고 24시간 동안 재배양하여 MUC5AC의 생성을 유도하였다. 그 다음, 배양 상등액만을 취해 MUC5AC 분비량을 측정 진행하였다.Afterwards, the cells were treated with PMA at a concentration of 100 ng/mL and re-cultured for 24 hours to induce the production of MUC5AC. Next, only the culture supernatant was taken and the secretion amount of MUC5AC was measured.
도 5에서 보는 바와 같이, PMA에 의해 유도된 MUC5AC의 분비가 감태추출물혼합물에 의해 농도 의존적으로 억제됨을 확인하였다. 또한 100μg/mL의 동일한 농도에서 감태, 감국추출물과 각각 비교하였을 때, 감태추출물에 비하여 염증성 점액단백질 생성 억제능이 약 100% 증가하고, 100 ㎍/mL 농도의 감국추출물에 비하여, 염증성 점액단백질 생성 억제능이 50% 증가함으로써, 태감국 복합물의 MUC5AC의 분비 억제능이 현저하게 향상되는 것으로 확인되었다.As shown in Figure 5, it was confirmed that the secretion of MUC5AC induced by PMA was suppressed in a concentration-dependent manner by the Ecklonia cava extract mixture. Additionally, when compared with Ecklonia cava and chrysanthemum chrysanthemum extracts at the same concentration of 100 μg/mL, the ability to inhibit the production of inflammatory mucus proteins increases by about 100% compared to the Ecklonia chrysanthemum extract, and compared to the Chrysanthemum chrysanthemum extract at a concentration of 100 μg/mL, the ability to inhibit the production of inflammatory mucus proteins increases by about 100%. By increasing this by 50%, it was confirmed that the MUC5AC secretion inhibitory ability of the Taegam Guk complex was significantly improved.
해당 결과를 통하여 감국추출물은 과도한 가래 분비를 억제할 수 있음을 확인하였다.The results confirmed that Chrysanthemum chrysanthemum extract can suppress excessive phlegm secretion.
이상으로 본 발명의 내용의 특정한 부분을 상세히 기술하였는 바, 당업계의 통상의 지식을 가진 자에게 있어서, 이러한 구체적 기술은 단지 바람직한 실시예일 뿐이며, 이에 의해 본 발명의 범위가 제한되는 것이 아닌 점은 명백할 것이다.As the specific parts of the present invention have been described in detail above, those skilled in the art will understand that these specific techniques are merely preferred embodiments and do not limit the scope of the present invention. It will be obvious.
따라서, 본 발명의 실질적인 범위는 첨부된 청구항들과 그것들의 등가물에 의하여 정의된다고 할 것이다. 본 발명의 단순한 변형 내지 변경은 이 분야의 통상의 지식을 가진 자에 의하여 용이하게 이용될 수 있으며, 이러한 변형이나 변경은 모두 본 발명의 영역에 포함되는 것으로 볼 수 있다. Accordingly, the substantial scope of the present invention will be defined by the appended claims and their equivalents. Simple modifications or changes of the present invention can be easily used by those skilled in the art, and all such modifications or changes can be considered to be included in the scope of the present invention.
Claims (13)
상기 감태추출물혼합물의 농도 100 ㎍/mL에서,
100 ㎍/mL 농도의 감태추출물에 비하여, 염증성 점액단백질 생성 억제능이 90~110% 증가하고,
100 ㎍/mL 농도의 감국추출물에 비하여, 염증성 점액단백질 생성 억제능이 45~55% 증가하는 것을 특징으로 하는 염증성 호흡기 질환 예방 또는 개선용 건강기능식품 조성물.
A health functional food composition for preventing or improving inflammatory respiratory diseases comprising an Ecklonia cava extract mixture containing Ecklonia cava extract and Chrysanthemum chrysanthemum extract as an active ingredient,
At a concentration of 100 μg/mL of the Ecklonia cava extract mixture,
Compared to Ecklonia cava extract at a concentration of 100 ㎍/mL, the ability to inhibit inflammatory mucus protein production increases by 90-110%,
A health functional food composition for preventing or improving inflammatory respiratory diseases, characterized in that the ability to inhibit inflammatory mucus protein production increases by 45 to 55% compared to Chrysanthemum chrysanthemum extract at a concentration of 100 ㎍/mL.
The health functional food composition for preventing or improving inflammatory respiratory disease according to claim 1, wherein the Ecklonia cava extract mixture contains the Ecklonia cava extract and the Chrysanthemum chrysanthemum extract in a weight ratio of 10 to 90:10 to 90.
The health functional food composition for preventing or improving inflammatory respiratory disease according to claim 1, wherein the Ecklonia cava extract mixture contains the Ecklonia cava extract and the Chrysanthemum chrysanthemum extract in a weight ratio of 50:50.
The health functional food composition for preventing or improving inflammatory respiratory disease according to claim 1, wherein the concentration of the Ecklonia cava extract mixture is 90 to 110 μg/mL.
The health function for preventing or improving inflammatory respiratory diseases according to claim 1, wherein the respiratory disease is one or more selected from the group consisting of chronic obstructive pulmonary disease (COPD), asthma, pharyngitis, cough, phlegm, bronchitis, and pneumonia. Food composition.
상기 감태추출물혼합물의 농도 100 ㎍/mL에서,
100 ㎍/mL 농도의 감태추출물에 비하여, 염증성 점액단백질 생성 억제능이90~110% 증가하고,
100 ㎍/mL 농도의 감국추출물에 비하여, 염증성 점액단백질 생성 억제능이 45~55% 증가하는 것을 특징으로 하는 염증성 호흡기 질환 예방 또는 치료용 약학적 조성물.
A pharmaceutical composition for preventing or treating inflammatory respiratory diseases comprising an Ecklonia cava extract mixture containing Ecklonia cava extract and Chrysanthemum chrysanthemum extract as an active ingredient,
At a concentration of 100 μg/mL of the Ecklonia cava extract mixture,
Compared to Ecklonia cava extract at a concentration of 100 ㎍/mL, the ability to inhibit inflammatory mucus protein production increases by 90-110%,
A pharmaceutical composition for preventing or treating inflammatory respiratory diseases, characterized in that the ability to inhibit inflammatory mucus protein production is increased by 45-55% compared to the Chrysanthemum chrysanthemum extract at a concentration of 100 ㎍/mL.
The pharmaceutical composition for preventing or treating inflammatory respiratory disease according to claim 7, wherein the Ecklonia cava extract mixture contains the Ecklonia cava extract and the Chrysanthemum chrysanthemum extract in a weight ratio of 10 to 90:10 to 90.
The pharmaceutical composition for preventing or treating inflammatory respiratory diseases according to claim 7, wherein the Ecklonia cava extract mixture contains the Ecklonia cava extract and the Chrysanthemum chrysanthemum extract in a weight ratio of 50:50.
The pharmaceutical composition for preventing or treating inflammatory respiratory diseases according to claim 7, wherein the concentration of the Ecklonia cava extract mixture is 90 to 110 μg/mL.
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