KR20220033826A - Composition for preventing or treating chronic rhinosinusitis comprising Ginsenoside F1 - Google Patents
Composition for preventing or treating chronic rhinosinusitis comprising Ginsenoside F1 Download PDFInfo
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- KR20220033826A KR20220033826A KR1020200116158A KR20200116158A KR20220033826A KR 20220033826 A KR20220033826 A KR 20220033826A KR 1020200116158 A KR1020200116158 A KR 1020200116158A KR 20200116158 A KR20200116158 A KR 20200116158A KR 20220033826 A KR20220033826 A KR 20220033826A
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- KR
- South Korea
- Prior art keywords
- ginsenoside
- chronic sinusitis
- sinusitis
- composition
- chronic
- Prior art date
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- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
Description
본 발명은 진세노사이드 F1을 포함하는 만성 부비동염 예방 또는 치료용 조성물 및 만성 부비동염 예방 또는 치료 방법에 관한 것이다.The present invention relates to a composition for preventing or treating chronic sinusitis comprising ginsenoside F1 and a method for preventing or treating chronic sinusitis.
만성 부비동염은 부비동 점막 비후를 초래하는 질병으로서 도시 인구 중 5~15%의 발생 빈도를 보일만큼 가장 흔한 질환 중 하나이다. 만성 부비동염은 과거에 단순 감염 질환으로 취급되었으나, 다양한 연구에 의하여 단순히 세균, 바이러스, 진균, 등에 의한 감염 질환이 아닌 여러 인자에 의한 만성 질환임이 밝혀졌다. Chronic sinusitis is a disease that causes sinus mucosal thickening and is one of the most common diseases with an incidence of 5-15% of the urban population. Chronic sinusitis was treated as a simple infectious disease in the past, but various studies have revealed that it is a chronic disease caused by various factors, not simply an infectious disease caused by bacteria, viruses, fungi, and the like.
이렇듯 만성 부비동염은 발병기전이 복잡하며 모호할 뿐만 아니라, 상당한 보건학 및 사회학적 비용을 유발함에도 불구하고 아직까지 항생제 및 스테로이드제 등 대중적인 치료법 외에 병의 재발을 막는 궁극적인 치료법이 개발되지 않은 실정이다.As such, the pathogenesis of chronic sinusitis is complex and ambiguous, and despite the fact that it incurs significant health and social costs, an ultimate treatment for preventing recurrence of the disease has not yet been developed other than popular treatments such as antibiotics and steroids. .
한편, 진세노사이드는 인삼에 있는 사포닌으로, 30종 이상의 다양한 종류가 존재하는데, 구조에 따라 PPD(protopanaxadiol) 계열과 PPT(protopanaxatriol) 계열로 나뉘며, 화학적 구조에 따라 각기 다른 약리활성을 나타내는 것으로 알려져 있다(Curr Vasc Pharmacol. 2009 July;7(3):293-302). 그러나, 상기 진세노사이드 중 분리가 어려운 진세노사이드의 만성 부비동염 치료 효과에 대해서는 보고된 바 없다.On the other hand, ginsenoside is a saponin in ginseng, and there are more than 30 different types. According to the structure, it is divided into a PPD (protopanaxadiol) series and a PPT (protopanaxatriol) series, and is known to exhibit different pharmacological activities depending on the chemical structure. ( Curr Vasc Pharmacol. 2009 July;7(3):293-302). However, there has been no report on the therapeutic effect of chronic sinusitis of ginsenoside, which is difficult to separate among the ginsenosides.
이러한 배경하에, 본 발명자들은 일반적인 만성 부비동염을 근본적으로 치료하기 위해 예의 노력한 결과, 진세노사이드 F1이 만성 부비동염에서 더 나아가 호산구성 만성 부비동염, 및 난치성 부비동염까지 근본적으로 치료할 수 있음을 확인함으로써 본 발명을 완성하였다.Under this background, the present inventors have made intensive efforts to fundamentally treat common chronic sinusitis, and as a result, ginsenoside F1 can fundamentally treat chronic sinusitis, eosinophilic sinusitis, and refractory sinusitis. completed.
본 발명의 하나의 목적은 진세노사이드 F1을 유효성분으로 포함하는 만성 부비동염 예방 또는 치료용 약학적 조성물을 제공하는 것이다.One object of the present invention is to provide a pharmaceutical composition for preventing or treating chronic sinusitis comprising ginsenoside F1 as an active ingredient.
본 발명의 다른 하나의 목적은 진세노사이드 F1을 유효성분으로 포함하는 만성 부비동염 예방 또는 개선용 식품 조성물을 제공하는 것이다.Another object of the present invention is to provide a food composition for preventing or improving chronic sinusitis comprising ginsenoside F1 as an active ingredient.
본 발명의 또 다른 하나의 목적은 진세노사이드 F1을 유효성분으로 포함하는 만성 부비동염 예방 또는 개선용 사료 조성물을 제공하는 것이다.Another object of the present invention is to provide a feed composition for preventing or improving chronic sinusitis comprising ginsenoside F1 as an active ingredient.
본 발명의 또 다른 하나의 목적은 만성 부비동염 예방 또는 치료 방법을 제공하는 것이다.Another object of the present invention is to provide a method for preventing or treating chronic sinusitis.
이하, 본 발명 내용에 대하여 구체적으로 설명하면 다음과 같다. 한편, 본 발명에서 개시한 일 양태의 설명 및 실시형태는 공통된 사항에 대하여 다른 양태의 설명 및 실시 형태에도 적용될 수 있다. 또한, 본 발명에서 개시된 다양한 요소들의 모든 조합이 본 발명의 범주에 속한다. 더불어, 하기 기술된 구체적인 서술에 의하여 본 발명의 범주가 제한된다고 볼 수 없다.Hereinafter, the content of the present invention will be described in detail as follows. On the other hand, descriptions and embodiments of one aspect disclosed in the present invention may be applied to descriptions and embodiments of other aspects with respect to common matters. Also, all combinations of the various elements disclosed herein are within the scope of the present invention. In addition, it cannot be seen that the scope of the present invention is limited by the specific descriptions described below.
상술한 목적을 달성하기 위한 본 발명의 일 양태는 진세노사이드 F1을 유효성분으로 포함하는 만성 부비동염(chronic rhinosinusitis; CRS) 예방 또는 치료용 약학적 조성물을 제공한다. 구체적으로, 상기 만성 부비동염은 호산구성 만성 부비동염 또는 난치성 부비동염일 수 있으나, 이에 제한되지 않는다.One aspect of the present invention for achieving the above object provides a pharmaceutical composition for preventing or treating chronic sinusitis (CRS) comprising ginsenoside F1 as an active ingredient. Specifically, the chronic sinusitis may be eosinophilic chronic sinusitis or intractable sinusitis, but is not limited thereto.
본 발명의 용어 "만성 부비동염"(chronic rhinosinusitis; CRS)은 부비동염의 일종으로서, 부비동 점막 비후나 부비동 혼탁이 초래된 만성 염증성 병변을 의미하며, 세균, 바이러스, 진균 등에 의한 감염 질환이 아니라 여러 인자에 의한 만성 염증 질환이다. 부비동염은 만성 부비동염과 급성 부비동염으로 구분되며, 부비동염은 비염을 동반하기도 하므로 비부비동염과 혼용될 수 있다. 구체적으로, 상기 만성 부비동염은 호산구성 만성 부비동염, 난치성 부비동염일 수 있으나, 이에 제한되지 않는다. As used herein, the term "chronic rhinosinusitis" (CRS) is a type of sinusitis, and refers to a chronic inflammatory lesion that results in sinus mucosal thickening or sinus clouding, and is not caused by bacterial, viral, fungal infection, but by several factors. It is a chronic inflammatory disease caused by Sinusitis is divided into chronic sinusitis and acute sinusitis, and sinusitis can be mixed with rhinosinusitis because it also accompanies rhinitis. Specifically, the chronic sinusitis may be eosinophilic chronic sinusitis or intractable sinusitis, but is not limited thereto.
본 발명의 용어 "호산구성 만성 부비동염"은 호산구 증가의 염증반응이 깊이 관여하는 만성 부비동염을 의미한다. 만성 부비동염은 호산구성 및 비호산구성 만성 부비동염으로 구분되며, 호산구 관여 정도, 말초혈액 내의 호산구 수, 및 증상 등에 의해 구분될 수 있다. 예를 들어, 호산구성 만성 부비동염의 경우 혈중 호산구가 비강 점막에 침윤하여 사골동(ethmoid sinus) 및 후각열(olfactory cleft) 점막을 붓게 하는 경향이 있다. 또한, 알레르기성 뮤신과 같이 샛노란 젤리같은 분비물을 보이는 것이 특징이며, 후각장애가 먼저 동반된다. 반면, 비호산구성 만성 부비동염의 경우 염증의 시작은 국소 반응으로서, 자극받은 곳이 먼저 염증이 생기고 비강 내 공기의 흐름이 개구비도 단위(Ostiomeatal unit)를 통하게 되므로 이 부위의 점막이 붓게 되어 상악동의 입구를 막으면서 상악동에 농이 고이는 형태로 진행되므로, 개구비도 단위의 질환이라고도 할 수 있다. 상악동에 편재되어 병변을 보이게 되며, 점액농이 분비되는 것이 특징이다. 또한, 호산구성 및 비호산구성 만성 부비동염 각각의 용종의 경우에도 색, 투명도, 질감, 형상, 및 조직학적으로 차이가 있어 구분되고 있다. As used herein, the term “chronic eosinophilic sinusitis” refers to chronic sinusitis in which the inflammatory response of increased eosinophils is deeply involved. Chronic sinusitis is divided into eosinophilic and non-eosinophilic chronic sinusitis, and can be classified by the degree of involvement of eosinophils, the number of eosinophils in the peripheral blood, and symptoms. For example, in the case of chronic eosinophilic sinusitis, blood eosinophils tend to infiltrate the nasal mucosa, causing swelling of the ethmoid sinus and olfactory cleft mucosa. In addition, it is characterized by the appearance of yellow jelly-like secretions like allergic mucin, accompanied by olfactory disorders first. On the other hand, in the case of non-eosinophilic chronic sinusitis, the onset of inflammation is a local reaction. The irritated area becomes inflamed first, and the air flow in the nasal cavity passes through the Ostioeatal unit, so the mucous membrane of this area swells and the maxillary sinus becomes inflamed. As it progresses in the form of accumulation of pus in the maxillary sinus while blocking the entrance of It is ubiquitous in the maxillary sinus and causes lesions, and is characterized by secretion of mucus. In addition, in the case of polyps of each of eosinophilic and non-eosinophilic chronic sinusitis, there are differences in color, transparency, texture, shape, and histology, and thus, they are distinguished.
본 발명의 용어 "난치성 부비동염"은 일반적인 만성 부비동염 치료제인 항생제, 염증 억제제, 또는 스테로이드제 등에 의해서도 치료가 불가능하거나 재발되는 부비동염을 의미한다. 부비동염 치료를 위해 1차적으로 염증 억제제 등이 사용되나 이는 근본적인 치료법이라고 할 수 없으며 재발하는 경우가 다수 있으므로, 본 발명은 일반적인 치료제와는 다른 경로의 유효성분을 개발하였다.As used herein, the term “refractory sinusitis” refers to sinusitis that cannot be treated or that recurs even with antibiotics, inflammation inhibitors, or steroids, which are general therapeutic agents for chronic sinusitis. Inflammatory inhibitors are primarily used for the treatment of sinusitis, but this cannot be called a fundamental treatment and there are many cases of recurrence. Therefore, the present invention has developed an active ingredient that is different from general therapeutic agents.
본 발명의 용어 "진세노사이드 F1"는 인삼 또는 홍삼 등의 삼의 주요 활성성분인 사포닌의 한 종류로서, PPT(protopanaxatriol) 계열에 속하는 진세노사이드를 의미한다. As used herein, the term “ginsenoside F1” is a kind of saponin, which is the main active ingredient of ginseng, such as ginseng or red ginseng, and refers to ginsenoside belonging to the PPT (protopanaxatriol) family.
구체적으로, 본 발명에서는 진세노사이드 F1을 유효성분으로 포함하는 조성물이 만성 부비동염을 예방 및 치료할 수 있음을 확인하였다(실시예 2-1 내지 2-4). 또한, 본 발명에서는 진세노사이드 F1을 유효성분으로 포함하는 조성물이 만성 부비동염 동물모델에서 호산구 농도를 크게 감소시켜 호산구성 만성 부비동염을 예방 및 치료할 수 있음을 확인하였다(실시예 2-2 , 2-3, 및 2-5, 및 도 2, 3, 및 8). 이는 진세노사이드 F1을 유효성분으로 포함하는 조성물이 만성 부비동염을 예방 및 치료할 수 있음을 시사하는 것이다. Specifically, in the present invention, it was confirmed that the composition comprising ginsenoside F1 as an active ingredient can prevent and treat chronic sinusitis (Examples 2-1 to 2-4). In addition, in the present invention, it was confirmed that the composition comprising ginsenoside F1 as an active ingredient can prevent and treat eosinophilic chronic sinusitis by significantly reducing the concentration of eosinophils in an animal model of chronic sinusitis (Examples 2-2, 2- 3, and 2-5, and FIGS. 2, 3, and 8). This suggests that the composition comprising ginsenoside F1 as an active ingredient can prevent and treat chronic sinusitis.
본 발명의 목적상 진세노사이드 F1은 자연살해세포(Natural killer cell; NK cell)의 활성을 촉진시키는 것일 수 있으며, 사이토카인 IFN-γ의 발현을 증가시키는 것일 수 있다.For the purpose of the present invention, ginsenoside F1 may promote the activity of natural killer cells (NK cells), and may increase the expression of cytokine IFN-γ.
본 발명의 용어 "자연살해세포"(Natural killer cell; NK cell)는 선천면역을 담당하는 세포 중 하나이며, 바이러스 감염 세포나 암 세포를 공격하는 기능이 있다고 알려져 있다. 상기 자연살해세포의 활성 촉진은 자연살해세포의 탈과립 활성, 세포살해 활성, 또는 세포살해인자의 발현이 증가되 세포용해능력이 증가일 수 있으며, 구체적으로 IFN-γ와 같은 사이토카인의 발현이 증가되는 것일 수 있으나, 이에 제한되지 않는다.As used herein, the term "natural killer cell" (Natural killer cell; NK cell) is one of the cells responsible for innate immunity and is known to have a function of attacking virus-infected cells or cancer cells. The promotion of the activity of the natural killer cells may be an increase in the cytolytic ability while the degranulation activity of the natural killer cells, the cell killing activity, or the expression of the cell killing factor is increased, specifically, the expression of cytokines such as IFN-γ is increased may be, but is not limited thereto.
구체적으로, 본 발명에서 만성 부비동염 동물모델에 일반적인 염증 억제제인 덱사메타손을 투여하였을 때 IFN-γ의 발현에 유의한 차이가 없음을 확인하였으나, 만성 부비동염 동물모델에 진세노사이드 F1을 투여한 결과, 알러지성 염증과 관련된 사이토카인뿐만 아니라, 자연살해세포에서 많이 발현하는 것으로 알려진 IFN-γ의 발현이 현저하게 증가하는 것을 확인하였다(실시예 2-4). 또한, 일반적인 염증 억제제인 덱사메타손에 비해 만성 부비동염 동물모델이 자연살해세포와 높은 연관성이 있음을 확인하였다(실시예 2-5). 이러한 결과는, 본 발명의 진세노사이드 F1이 일반적인 염증 억제제와는 전혀 다른 경로로 만성 부비동염을 예방 또는 치료함을 확인하여, 일반적인 만성 부비동염 뿐만 아니라, 호산구성 만성 부비동염, 난치성 부비동염, 특히, 염증 억제제 또는 항생제에 의해 치료가 불가능하거나 재발하는 난치성 부비동염까지 근본적으로 치료할 수 있는 우수한 효과가 있음을 시사하는 것이다.Specifically, it was confirmed that there was no significant difference in the expression of IFN-γ when dexamethasone, a general inhibitor of inflammation, was administered to an animal model of chronic sinusitis in the present invention, but as a result of administering ginsenoside F1 to an animal model of chronic sinusitis, allergy It was confirmed that the expression of IFN-γ, which is known to be widely expressed in natural killer cells, as well as cytokines related to sexual inflammation, significantly increased (Example 2-4). In addition, it was confirmed that the chronic sinusitis animal model had a higher correlation with natural killer cells compared to dexamethasone, a general inhibitor of inflammation (Example 2-5). These results confirm that the ginsenoside F1 of the present invention prevents or treats chronic sinusitis by a completely different route from that of a general anti-inflammatory agent, so that not only general chronic sinusitis, but also eosinophilic chronic sinusitis, intractable sinusitis, in particular, an inhibitor of inflammation Or, it suggests that there is an excellent effect that can fundamentally treat even intractable sinusitis that cannot be treated with antibiotics or that recurs.
본 발명에서 용어, "예방"은 본 발명의 조성물의 투여로 만성 부비동염의 발병 또는 진행을 억제 또는 지연시키는 모든 행위를 의미한다.As used herein, the term “prevention” refers to any action that inhibits or delays the onset or progression of chronic sinusitis by administration of the composition of the present invention.
본 발명에서 용어, "치료"는 치료하고자 하는 대상체 또는 세포의 천연 과정을 변경시키기 위해 임상적으로 개입하는 것을 지칭하고, 이는 임상 병리 상태가 진행되는 동안 또는 이를 예방하기 위해 수행할 수 있다. 목적하는 치료 효과에는 질병의 발생 또는 재발을 예방하고, 증상을 완화시키며, 질병에 따른 모든 직접 또는 간접적인 병리학적 결과를 저하시키며, 질병 진행 속도를 감소시키며, 질병 상태를 경감 또는 일시적 완화시키며, 차도시키거나 예후를 개선시키는 것이 포함된다. 구체적으로 본 발명의 조성물의 투여로 만성 부비동염을 호전시키는 모든 행위를 포함한다. As used herein, the term "treatment" refers to clinical intervention to alter the natural process of a subject or cell to be treated, which can be performed during or to prevent clinical pathology. The desired therapeutic effect includes preventing the occurrence or recurrence of a disease, alleviating symptoms, reducing any direct or indirect pathological consequences of the disease, reducing the rate of disease progression, alleviating or temporarily ameliorating the disease state; remission or improving the prognosis. Specifically, it includes all actions for alleviating chronic sinusitis by administering the composition of the present invention.
상기 약학 조성물은 약학적으로 허용 가능한 담체를 포함할 수 있다. 상기 "약학적으로 허용 가능한 담체"는 생물체를 자극하지 않으면서, 주입되는 화합물의 생물학적 활성 및 특성을 저해하지 않는 담체, 부형제 또는 희석제를 의미할 수 있으며, 구체적으로, 비자연적 담체(non-naturally occuring carrier)일 수 있다. 본 발명에 사용 가능한 상기 담체의 종류는 특별히 제한되지 아니하며 당해 기술 분야에서 통상적으로 사용되고 약학적으로 허용되는 담체라면 어느 것이든 사용할 수 있다. 상기 담체의 비제한적인 예로는, 식염수, 멸균수, 링거액, 완충 식염수, 알부민 주사 용액, 덱스트로스 용액, 말토 덱스트린 용액, 글리세롤, 에탄올 등을 들 수 있다. 이들은 단독으로 사용되거나 2 종 이상을 혼합하여 사용될 수 있다.The pharmaceutical composition may include a pharmaceutically acceptable carrier. The "pharmaceutically acceptable carrier" may refer to a carrier, excipient or diluent that does not inhibit the biological activity and properties of the injected compound without stimulating the organism, and specifically, non-naturally occurring carrier). The type of carrier usable in the present invention is not particularly limited, and any carrier commonly used in the art and pharmaceutically acceptable may be used. Non-limiting examples of the carrier include saline, sterile water, Ringer's solution, buffered saline, albumin injection solution, dextrose solution, maltodextrin solution, glycerol, ethanol, and the like. These may be used alone or in mixture of two or more.
약학적으로 허용 가능한 담체를 포함하는 상기 조성물은 경구 또는 비경구의 여러 가지 제형일 수 있다. 제제화할 경우에는 보통 사용하는 충진제, 증량제, 결합제, 습윤제, 붕해제, 계면활성제 등의 희석제 또는 부형제를 사용하여 조제된다.The composition comprising a pharmaceutically acceptable carrier may be in various oral or parenteral formulations. In the case of formulation, it is prepared using commonly used diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrants, and surfactants.
상세하게는, 경구투여를 위한 고형제제에는 정제, 환제, 산제, 과립제, 캡슐제 등이 포함되며, 이러한 고형제제는 상기 화합물에 적어도 하나 이상의 부형제, 예를 들면, 전분, 칼슘카보네이트, 수크로오스, 락토오스, 젤라틴 등을 섞어 조제될 수 있다. 또한, 단순한 부형제 이외에 마그네슘 스테아레이트, 탈크 같은 윤활제들도 사용될 수 있다. 경구를 위한 액상 제제로는 현탁제, 내용액제, 유제, 시럽제 등이 해당되는데, 흔히 사용되는 단순 희석제인 물, 액체 파라핀 이외에 여러 가지 부형제, 예를 들면 습윤제, 감미제, 방향제, 보존제 등이 포함될 수 있다. 비경구 투여를 위한 제제에는 멸균된 수용액, 비수성용제, 현탁제, 유제, 동결건조 제제 및 좌제가 포함된다. 비수성용제, 현탁제로는 프로필렌글리콜, 폴리에틸렌 글리콜, 올리브 오일과 같은 식물성 오일, 에틸올레이트와 같은 주사 가능한 에스테르 등이 사용될 수 있다. 좌제의 기제로는 위텝솔, 마크로골, 트윈 61, 카카오지, 라우린지, 글리세로젤라틴 등이 사용될 수 있다.Specifically, solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc., and these solid preparations include at least one excipient to the compound, for example, starch, calcium carbonate, sucrose, lactose. , gelatin, etc. may be mixed and prepared. In addition to simple excipients, lubricants such as magnesium stearate and talc may also be used. Liquid formulations for oral use include suspensions, solutions, emulsions, and syrups. In addition to water and liquid paraffin, which are commonly used simple diluents, various excipients such as wetting agents, sweeteners, fragrances, and preservatives may be included. there is. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, lyophilized formulations and suppositories. Non-aqueous solvents and suspending agents include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. As the base of the suppository, Witepsol, Macrogol, Tween 61, cacao butter, laurin fat, glycerogelatin, etc. can be used.
또한, 본 발명의 약학 조성물은 약학적으로 유효한 양의 진세노사이드 F1을 포함할 수 있다. 본 발명에서 용어, "약학적으로 유효한 양"은 의학적 치료에 적용 가능한 합리적인 수혜/위험 비율로 질환을 치료하기에 충분한 양을 의미하며, 일반적으로 0.001 내지 1000 mg/kg의 양, 구체적으로는 0.05 내지 200 mg/kg, 보다 구체적으로는 0.1 내지 100 mg/kg의 양, 더욱 구체적으로는 0.1 내지 50 mg/kg의 양을 일일 1회 내지 수회로 나누어 투여할 수 있다. 그러나 본 발명의 목적상, 특정 환자에 대한 구체적인 치료적 유효량은 달성하고자 하는 반응의 종류와 정도, 경우에 따라 다른 제제가 사용되는지의 여부를 비롯한 구체적 조성물, 환자의 연령, 체중, 일반 건강 상태, 성별 및 식이, 투여 시간, 투여 경로 및 조성물의 분비율, 치료기간, 구체적 조성물과 함께 사용되거나 동시 사용되는 약물을 비롯한 다양한 인자와 의약 분야에 잘 알려진 유사 인자에 따라 다르게 적용하는 것이 바람직하다.In addition, the pharmaceutical composition of the present invention may contain a pharmaceutically effective amount of ginsenoside F1. As used herein, the term "pharmaceutically effective amount" means an amount sufficient to treat a disease at a reasonable benefit/risk ratio applicable to medical treatment, and is generally in an amount of 0.001 to 1000 mg/kg, specifically 0.05 to 200 mg/kg, more specifically, in an amount of 0.1 to 100 mg/kg, more specifically, in an amount of 0.1 to 50 mg/kg, may be administered once to several times a day. However, for the purposes of the present invention, a specific therapeutically effective amount for a particular patient depends on the type and extent of the response to be achieved, the specific composition, including whether other agents are used, if necessary, the specific composition, the patient's age, weight, general health, It is preferable to apply differently depending on various factors including sex and diet, administration time, administration route and secretion rate of the composition, treatment period, drugs used together or concurrently with a specific composition, and similar factors well known in the pharmaceutical field.
본 발명의 약학 조성물은 매일 투여 또는 간헐적으로 투여할 수 있고, 1일당 투여 횟수는 1회 또는 2~3회로 나누어 투여하는 것이 가능하다. 또한, 본 발명의 약학 조성물은 단독으로, 또는 다른 약물 치료와 병용하여 사용할 수 있다. 상기 요소를 모두 고려하여 부작용 없이 최소한의 양으로 최대 효과를 얻을 수 있는 양을 투여하는 것이 중요하며, 당업자에 의해 용이하게 결정될 수 있다The pharmaceutical composition of the present invention may be administered daily or intermittently, and the number of administrations per day may be administered once or divided into two to three times. In addition, the pharmaceutical composition of the present invention may be used alone or in combination with other drug treatments. Taking all of the above factors into consideration, it is important to administer an amount that can obtain the maximum effect with a minimum amount without side effects, and can be easily determined by those skilled in the art.
본 발명의 목적을 달성하기 위한 다른 하나의 양태는 진세노사이드 F1을 유효성분으로 포함하는 만성 부비동염 예방 또는 개선용 식품 조성물을 제공한다. 본 발명의 용어 "진세노사이드 F1", "만성 부비동염", 및 "예방"은 전술한 바와 같다.Another aspect for achieving the object of the present invention provides a food composition for preventing or improving chronic sinusitis comprising ginsenoside F1 as an active ingredient. The terms "ginsenoside F1", "chronic sinusitis", and "prevention" of the present invention are as described above.
본 발명에서 용어 “개선”은 본 발명의 진세노사이드 F1을 유효성분으로 포함하는 조성물의 투여로 만성 부비동염이 호전 또는 이롭게 변경되는 모든 행위를 의미한다.In the present invention, the term “improvement” refers to any action in which chronic sinusitis is improved or changed to advantage by administration of a composition comprising the ginsenoside F1 of the present invention as an active ingredient.
본 발명의 용어 "식품"은 육류, 소시지, 빵, 초콜릿, 캔디류, 스넥류, 과자류, 피자, 라면, 기타 면류, 껌류, 아이스크림류를 포함한 낙농제품, 각종 스프, 음료수, 차, 드링크제, 알코올음료, 비타민 복합체, 건강 기능 식품 및 건강 식품 등이 있으며, 통상적인 의미의 식품을 모두 포함한다.As used herein, the term “food” refers to meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, dairy products including ice cream, various soups, beverages, tea, drinks, alcoholic beverages, There are vitamin complexes, health functional foods, and health foods, and includes all foods in the ordinary sense.
상기 건강기능(성) 식품(functional food)이란, 특정보건용 식품(food for special health use, FoSHU)와 동일한 용어로, 영양 공급 외에도 생체조절기능이 효율적으로 나타나도록 가공된 의학, 의료효과가 높은 식품을 의미한다. 여기서 "기능(성)"이라 함은 인체의 구조 및 기능에 대하여 영양소를 조절하거나 생리학적 작용 등과 같은 보건용도에 유용한 효과를 얻는 것을 의미한다. 본 발명의 식품은 당 업계에서 통상적으로 사용되는 방법에 의하여 제조가능하며, 상기 제조시에는 당 업계에서 통상적으로 첨가하는 원료 및 성분을 첨가하여 제조할 수 있다. 또한 상기 식품의 제형 또한 식품으로 인정되는 제형이면 제한 없이 제조될 수 있다. 본 발명의 식품용 조성물은 다양한 형태의 제형으로 제조될 수 있으며, 일반 약품과는 달리 식품을 원료로 하여 약품의 장기 복용 시 발생할 수 있는 부작용 등이 없는 장점이 있고, 휴대성이 뛰어나, 본 발명의 식품은 면역증진 효과를 증진시키기 위한 보조제로 섭취가 가능하다.The functional food (functional food) is the same term as food for special health use (FoSHU), and in addition to supplying nutrients, it is processed to efficiently exhibit bioregulatory functions and has high medical effects. means food. Here, "function (sex)" means to obtain a useful effect for health purposes such as regulating nutrients or physiological action with respect to the structure and function of the human body. The food of the present invention can be prepared by a method commonly used in the art, and at the time of manufacture, it can be prepared by adding raw materials and components commonly added in the art. In addition, the formulation of the food may be prepared without limitation as long as it is a formulation recognized as a food. The composition for food of the present invention can be prepared in various forms, and unlike general drugs, it has the advantage that there are no side effects that may occur during long-term administration of the drug using food as a raw material, and it is excellent in portability, and the present invention Foods of can be ingested as an adjuvant to enhance the immune-boosting effect.
상기 건강 식품(health food)은 일반식품에 비해 적극적인 건강유지나 증진 효과를 가지는 식품을 의미하고, 건강보조식품(health supplement food)는 건강보조 목적의 식품을 의미한다. 경우에 따라, 건강 기능 식품, 건강식품, 건강보조식품의 용어는 호용된다.The health food means a food having an active health maintenance or promotion effect compared to general food, and the health supplement food means a food for the purpose of health supplementation. In some cases, the terms health functional food, health food, and dietary supplement are preferred.
구체적으로, 상기 건강 기능 식품은 진세노사이드 F1을 음료, 차류, 향신료, 껌, 과자류 등의 식품소재에 첨가하거나, 캡슐화, 분말화, 현탁액 등으로 제조한 식품으로, 이를 섭취할 경우 건강상 특정한 효과를 가져오는 것을 의미하나, 일반 약품과는 달리 식품을 원료로 하여 약품의 장기 복용 시 발생할 수 있는 부작용이 없는 장점이 있다.Specifically, the health functional food is a food prepared by adding ginsenoside F1 to food materials such as beverages, teas, spices, gum, and confectionery, or encapsulating, powdering, suspension, etc., and when ingested, It means to bring an effect, but unlike general drugs, it has the advantage that there are no side effects that may occur when taking the drug for a long time using food as a raw material.
본 발명의 식품 조성물은, 일상적으로 섭취하는 것이 가능하기 때문에 높은 면역증진 효과를 기대할 수 있어 매우 유용하다.The food composition of the present invention is very useful because it can be ingested on a daily basis, and thus a high immune enhancing effect can be expected.
상기 조성물은 생리학적으로 허용 가능한 담체를 추가로 포함할 수 있는데, 담체의 종류는 특별히 제한되지 아니하며 당해 기술 분야에서 통상적으로 사용되는 담체라면 어느 것이든 사용할 수 있다.The composition may further include a physiologically acceptable carrier, the type of carrier is not particularly limited, and any carrier commonly used in the art may be used.
또한, 상기 조성물은 식품 조성물에 통상 사용되어 냄새, 맛, 시각 등을 향상시킬 수 있는 추가 성분을 포함할 수 있다. 예들 들어, 비타민 A, C, D, E, B1, B2, B6, B12, 니아신(niacin), 비오틴(biotin), 폴레이트(folate), 판토텐산(panthotenic acid) 등을 포함할 수 있다. 또한, 아연(Zn), 철(Fe), 칼슘(Ca), 크롬(Cr), 마그네슘(Mg), 망간(Mn), 구리(Cu), 크륨(Cr) 등의 미네랄을 포함할 수 있다. 또한, 라이신, 트립토판, 시스테인, 발린 등의 아미노산을 포함할 수 있다. In addition, the composition may include additional ingredients that are commonly used in food compositions to improve odor, taste, vision, and the like. For example, vitamins A, C, D, E, B1, B2, B6, B12, niacin, biotin, folate, pantothenic acid, and the like may be included. Also, it may include minerals such as zinc (Zn), iron (Fe), calcium (Ca), chromium (Cr), magnesium (Mg), manganese (Mn), copper (Cu), and chromium (Cr). In addition, it may include amino acids such as lysine, tryptophan, cysteine, and valine.
또한, 상기 조성물은 방부제(소르빈산 칼륨, 벤조산나트륨, 살리실산, 데히드로초산나트륨 등), 살균제(표백분과 고도 표백분, 차아염소산나트륨 등), 산화방지제(부틸히드록시아니졸(BHA), 부틸히드록시톨류엔(BHT) 등), 착색제(타르색소 등), 발색제(아질산 나트륨, 아초산 나트륨 등), 표백제(아황산나트륨), 조미료(MSG 글루타민산나트륨 등), 감미료(둘신, 사이클레메이트, 사카린, 나트륨 등), 향료(바닐린, 락톤류 등), 팽창제(명반, D-주석산수소칼륨 등), 강화제, 유화제, 증점제(호료), 피막제, 검기초제, 거품억제제, 용제, 개량제 등의 식품 첨가물(food additives)을 포함할 수 있다. 상기 첨가물은 식품의 종류에 따라 선별되고 적절한 양으로 사용될 수 있다.In addition, the composition includes a preservative (potassium sorbate, sodium benzoate, salicylic acid, sodium dehydroacetate, etc.), a disinfectant (bleaching powder and high bleaching powder, sodium hypochlorite, etc.), an antioxidant (butylhydroxyanisole (BHA), butylhydroxy toluene (BHT), etc.), coloring agents (tar pigments, etc.), coloring agents (sodium nitrite, sodium nitrite, etc.), bleach (sodium sulfite), seasonings (MSG sodium glutamate, etc.), sweeteners (dulcin, cyclamate, saccharin, Sodium, etc.), flavorings (vanillin, lactones, etc.), swelling agents (alum, D-potassium hydrogen tartrate, etc.), strengthening agents, emulsifiers, thickeners (flavors), film agents, gum base agents, foam inhibitors, solvents, food additives such as improving agents (food additives). The additive may be selected according to the type of food and used in an appropriate amount.
상기 진세노사이드 F1을 그대로 첨가하거나 다른 식품 또는 식품 성분과 함께 사용될 수 있고, 통상적인 방법에 따라 적절하게 사용될 수 있다. 유효성분의 혼합양은 그의 사용 목적(예방, 건강 또는 치료적 처치)에 따라 적합하게 결정될 수 있다. 일반적으로, 식품 또는 음료의 제조시에 본 발명의 식품 조성물은 식품 또는 음료에 대하여 50 중량부 이하, 구체적으로 20 중량부 이하의 양으로 첨가될 수 있다. 그러나 건강 및 위생을 목적으로 장기간 섭취할 경우에는 상기 범위 이하의 함량을 포함할 수 있으며, 안전성 면에서 아무런 문제가 없기 때문에 유효성분은 상기 범위 이상의 양으로도 사용될 수 있다.The ginsenoside F1 may be added as it is or may be used together with other foods or food ingredients, and may be appropriately used according to a conventional method. The mixed amount of the active ingredient may be suitably determined according to the purpose of its use (prevention, health or therapeutic treatment). In general, when preparing food or beverage, the food composition of the present invention may be added in an amount of 50 parts by weight or less, specifically 20 parts by weight or less with respect to the food or beverage. However, when ingested for a long period of time for health and hygiene purposes, it may contain an amount below the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount above the above range.
본 발명의 식품 조성물의 일 예로 건강음료 조성물으로 사용될 수 있으며, 이 경우 통상의 음료와 같이 여러 가지 향미제 또는 천연 탄수화물 등을 추가 성분으로 함유할 수 있다. 상술한 천연 탄수화물은 포도당, 과당과 같은 모노사카라이드; 말토스, 슈크로스와 같은 디사카라이드; 덱스트린, 사이클로덱스트린과 같은 폴리사카라이드; 자일리톨, 소르비톨, 에리트리톨 등의 당알콜일 수 있다. 감미제는 타우마틴, 스테비아 추출물과 같은 천연 감미제; 사카린, 아스파르탐과 같은 합성 감미제 등을 사용할 수 있다. 상기 천연 탄수화물의 비율은 본 발명의 조성물 100 mL 당 일반적으로 약 0.01 ~ 0.04 g, 구체적으로는 약 0.02 ~ 0.03 g 이다.As an example of the food composition of the present invention, it may be used as a health drink composition, and in this case, it may contain various flavoring agents or natural carbohydrates as an additional component like a conventional drink. The above-mentioned natural carbohydrates include monosaccharides such as glucose and fructose; disaccharides such as maltose and sucrose; polysaccharides such as dextrin and cyclodextrin; It may be a sugar alcohol such as xylitol, sorbitol, or erythritol. Sweeteners include natural sweeteners such as taumatine, stevia extract; A synthetic sweetener such as saccharin or aspartame may be used. The proportion of the natural carbohydrate is generally about 0.01 to 0.04 g, specifically, about 0.02 to 0.03 g per 100 mL of the composition of the present invention.
상기 외에 건강음료 조성물은 여러 가지 영양제, 비타민, 전해질, 풍미제, 착색제, 펙트산, 펙트산의 염, 알긴산, 알긴산의 염, 유기산, 보호성 콜로이드 증점제, pH 조절제, 안정화제, 방부제, 글리세린, 알코올 또는 탄산화제 등을 함유할 수 있다. 그 밖에 천연 과일주스, 과일주스 음료, 또는 야채 음료의 제조를 위한 과육을 함유할 수 있다. 이러한 성분은 독립적으로 또는 혼합하여 사용할 수 있다. 이러한 첨가제의 비율은 크게 중요하진 않지만 본 발명의 조성물 100 중량부당 0.01 ~ 0.1 중량부의 범위에서 선택되는 것이 일반적이다.In addition to the above, the health drink composition includes various nutrients, vitamins, electrolytes, flavoring agents, colorants, pectic acid, pectic acid salts, alginic acid, alginic acid salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, It may contain alcohol, a carbonation agent, or the like. In addition, it may contain the pulp for the production of natural fruit juice, fruit juice beverage, or vegetable beverage. These components may be used independently or in combination. Although the proportion of these additives is not critical, it is generally selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the composition of the present invention.
본 발명의 식품 조성물은 면역증진 효과를 나타낼 수 있다면 다양한 중량%로 포함할 수 있으나, 구체적으로 진세노사이드 F1을 식품 조성물의 총중량 대비 0.00001 내지 100 중량% 또는 0.01 내지 80 중량%로 포함할 수 있다.The food composition of the present invention may include various weight% as long as it can exhibit an immune enhancing effect, but specifically ginsenoside F1 may be included in an amount of 0.00001 to 100% by weight or 0.01 to 80% by weight relative to the total weight of the food composition. .
본 발명의 목적을 달성하기 위한 또 다른 하나의 양태는 진세노사이드 F1을 유효성분으로 포함하는 만성 부비동염 예방 또는 개선용 사료 조성물을 제공한다. 본 발명의 용어 "진세노사이드 F1", "만성 부비동염", "예방", 및 "개선"은 전술한 바와 같다.Another aspect for achieving the object of the present invention provides a feed composition for preventing or improving chronic sinusitis comprising ginsenoside F1 as an active ingredient. The terms "ginsenoside F1", "chronic sinusitis", "prevention", and "amelioration" of the present invention are as described above.
상기 사료 조성물에는 상기 진세노사이드 F1 외에도 약학적, 식품학적 또는 사료용으로 허용되는 공지의 담체, 안정화제, 또는 첨가제가 포함될 수 있다. 예를 들어, 품질 저하를 방지하기 위하여 첨가하는 결착제, 유화제, 보존제 등이 있고, 효용 증대를 위하여 사료에 첨가하는 아미노산제, 비타민제, 효소제, 향미제, 비단백질태질소화합물, 규산염제, 완충제, 추출제, 올리고당 등이 있다. 그 외에도 사료 혼합제 등을 추가로 포함할 수 있으며 이에 한정된 것은 아니다. 상기 사료 조성물은 필요에 따라 비타민, 아미노산류, 미네랄 등의 각종 양분, 항산화제 및 기타의 첨가제 등을 포함할 수도 있으며, 그 형상으로서는 분체, 과립, 펠릿, 현탁액 등의 적당한 상태일 수 있다. 본 발명의 사료 조성물은 단위 동물에 대하여 단독으로 또는 사료에 혼합하여 공급할 수 있다. 본 발명의 사료는 특별히 한정되는 것은 아니고, 분말 사료, 고형 사료, 건사료, 습식사료, 모이스트 펠릿 사료, 드라이 펠릿 사료, EP(Extruder Pellet) 사료, 날먹이 등 어떠한 사료라도 무방하다.In addition to the ginsenoside F1, the feed composition may include a known carrier, stabilizer, or additive acceptable for pharmaceutical, food, or feed. For example, there are binders, emulsifiers, and preservatives added to prevent quality deterioration, and amino acids, vitamins, enzymes, flavoring agents, non-protein nitrogenous compounds, silicate agents, buffers added to feed to increase efficacy. , extractants, and oligosaccharides. In addition, it may further include a feed mixture, and the like, but is not limited thereto. The feed composition may contain various nutrients such as vitamins, amino acids, minerals, antioxidants, and other additives as necessary, and may be in a suitable state such as powder, granule, pellet, suspension, etc. in its shape. The feed composition of the present invention may be supplied alone or mixed with feed for a unit animal. The feed of the present invention is not particularly limited, and any feed such as powder feed, solid feed, dry feed, wet feed, moist pellet feed, dry pellet feed, EP (Extruder Pellet) feed, raw feed, etc. may be used.
본 발명의 목적을 달성하기 위한 또 다른 하나의 양태는 상기 약학적 조성물을 인간을 제외한 개체에 투여하는 단계를 포함하는, 만성 부비동염 예방 또는 치료 방법을 제공한다. 본 발명의 용어 "약학적 조성물", "만성 부비동염", "예방", 및 "치료"는 전술한 바와 같다.Another aspect for achieving the object of the present invention provides a method for preventing or treating chronic sinusitis, comprising administering the pharmaceutical composition to an individual other than humans. The terms "pharmaceutical composition", "chronic sinusitis", "prevention", and "treatment" of the present invention are as described above.
본 발명의 용어 "투여"는 어떠한 적절한 방법으로 개체에게 본 발명의 조성물을 도입하는 것을 의미하며, 조성물의 투여 경로는 경구 또는 비경구의 다양한 경로를 통하여 투여될 수 있다. 구체적으로, 비경구 투여 시 비강 투여, 피부 외용 또는 복강 내 주사, 직장 내 주사, 피하주사, 정맥 내 주사, 근육 내 주사 또는 흉부 내 주사 등의 주입방식을 선택할 수 있다. 상기 비강 투여는 비강 내 흡입 및 비강 내 주사 등의 방식을 선택할 수 있으나, 이에 제한되지 않는다. 또한, 상기 조성물은 약학적으로 유효한 양으로 투여할 수 있다. The term "administration" of the present invention means introducing the composition of the present invention to an individual by any suitable method, and the administration route of the composition may be administered through various routes, either oral or parenteral. Specifically, for parenteral administration, an injection method such as nasal administration, external skin application or intraperitoneal injection, rectal injection, subcutaneous injection, intravenous injection, intramuscular injection, or intrathoracic injection may be selected. For the nasal administration, methods such as intranasal inhalation and intranasal injection may be selected, but the present invention is not limited thereto. In addition, the composition may be administered in a pharmaceutically effective amount.
본 발명의 용어 "개체"는 면역증진이 필요하거나 필요할 가능성이 있는 인간을 포함한 쥐, 생쥐, 가축 등의 모든 동물을 의미한다. 구체적으로, 인간을 포함한 포유동물일 수 있다.As used herein, the term “subject” refers to all animals, such as mice, mice, and livestock, including humans, in need or likely to require immune enhancement. Specifically, it may be a mammal including a human.
진세노사이드 F1에 의한 자연살해세포기반 면역활성 증진이 난치성 만성부비동염의 새로운 치료방안으로 활용될 수 있다. 이를 통해, 일반적인 만성 부비동염에서 더 나아가, 호산구성 만성 부비동염 및 난치성 부비동염까지 질병 치료에서의 산업적 활용을 기대할 수 있다.Natural killer cell-based immune activity enhancement by ginsenoside F1 can be used as a new treatment method for intractable chronic sinusitis. Through this, further from general chronic sinusitis, industrial application in disease treatment can be expected from eosinophilic chronic sinusitis and intractable sinusitis.
도 1은 만성 부비동염 동물모델의 제조 방법을 나타낸 모식도이다.
도 2는 진세노사이드 F1의 만성 부비동염 치료 효과를 나타낸 도이다.
도 3은 진세노사이드 F1과 일반적인 염증 억제제의 만성 부비동염 치료 효과를 비교하여 나타낸 도이다.
도 4는 진세노사이드 F1과 일반적인 염증 억제제의 만성 부비동염 치료 효과를 비교하여 나타낸 도이다.
도 5는 진세노사이드 F1이 염증 사이토카인 발현 변화에 미치는 영향을 나타낸 도이다.
도 6은 자연살해세포가 결핍된 만성 부비동염 동물모델 제조 방법을 나타낸 모식도이다.
도 7은 만성 부비동염 동물모델에서 자연살해세포가 결핍시킨 결과를 나타낸 도이다.
도 8은 진세노사이드 F1 및 일반적인 염증 억제제를 대상으로 자연살해세포가 결핍된 만성 부비동염 치료 효과를 비교하여 나타낸 도이다.1 is a schematic diagram showing a method of manufacturing an animal model of chronic sinusitis.
2 is a diagram showing the treatment effect of chronic sinusitis of ginsenoside F1.
3 is a view showing a comparison of the treatment effect of chronic sinusitis of ginsenoside F1 and a general inhibitor of inflammation.
Figure 4 is a diagram showing a comparison of the treatment effect of chronic sinusitis of ginsenoside F1 and a general inhibitor of inflammation.
5 is a diagram showing the effect of ginsenoside F1 on the expression of inflammatory cytokines.
6 is a schematic diagram showing a method for preparing an animal model of chronic sinusitis lacking natural killer cells.
7 is a diagram showing the results of the lack of natural killer cells in the chronic sinusitis animal model.
8 is a diagram showing the comparison of the treatment effects of ginsenoside F1 and general inflammation inhibitors for chronic sinusitis deficient in natural killer cells.
이하 본 발명을 실시예를 통하여 보다 상세하게 설명한다. 그러나 이들 실시예는 본 발명을 예시적으로 설명하기 위한 것으로 본 발명의 범위가 이들 실시예에 국한되는 것은 아니다.Hereinafter, the present invention will be described in more detail through examples. However, these examples are for illustrative purposes only, and the scope of the present invention is not limited to these examples.
실시예 1: 만성 부비동염 동물모델 제작Example 1: Production of an animal model of chronic sinusitis
호산성 만성 부비동염을 반영하는 적합한 동물모델을 확립하기 위하여 다음과 같은 실험을 수행하였다.The following experiments were performed to establish a suitable animal model reflecting chronic eosinophilic sinusitis.
구체적으로, 호흡기 염증질환 동물모델인 Balb/c 마우스에 Aspergillus oryzae 유래의 단백질분해효소(Aspergillus oryzae protease)와 오브알부민(ovalbumin; OVA)을 다양한 용량으로 주 3회 5주간 비강 내 점적하였으며, 대조군 마우스에 PBS를 동일한 조건으로 비강 내 점적하였다.Specifically, aspergillus oryzae -derived protease ( Aspergillus oryzae protease) and ovalbumin (OVA) were intranasally instilled in
그 결과, 최종적으로 Aspergillus oryzae 유래의 단백질분해효소 0.7U와 오브알부민 75㎍을 동시에 비강 내로 점적하였을 때 치명적이지 않고 재현성 있는 만성 부비동염 마우스 모델이 구축됨을 확인하였으며(도 1), 이 경우 조직 내 염증소견과 면역세포인 호산구 및 비만세포(mast cell)의 침윤이 증가됨을 관찰하였다. 이에, 상기 결과로부터 상기 마우스 모델은 난치·재발성의 호산성 만성 부비동염을 반영하는 적합한 동물모델이 됨을 확인하였다.As a result, it was finally confirmed that a non-fatal and reproducible chronic sinusitis mouse model was constructed when 0.7U of Aspergillus oryzae -derived protease and 75 μg of ovalbumin were simultaneously instilled into the nasal cavity (Fig. 1), in which case tissue inflammation It was observed that the infiltration of immune cells, eosinophils and mast cells, was increased. Accordingly, from the above results, it was confirmed that the mouse model was a suitable animal model reflecting refractory/recurrent chronic eosinophilic sinusitis.
실시예 2: 진세노사이드 F1의 만성 부비동염 치료 유효성 평가Example 2: Evaluation of the treatment efficacy of ginsenoside F1 for chronic sinusitis
실시예 2-1: 만성 부비동염 치료 유효성 평가 방법Example 2-1: Method for evaluating the treatment efficacy of chronic sinusitis
만성 부비동염 치료 유효성을 평가하기 위해, 만성 부비동염 유무 및 병증을 면역세포이자 염증세포인 호산구 및 비만세포의 침윤 감소 여부와 조직 내 염증소견으로 검증하였으며, 이를 위하여 상피층 비후 정도, 점막 두께, 및 염증세포의 유입 정도를 측정하였다.To evaluate the treatment effectiveness of chronic sinusitis, the presence and condition of chronic sinusitis was verified by whether the infiltration of eosinophils and mast cells, which are immune cells and inflammatory cells, decreased, and the inflammatory findings in the tissue. The degree of inflow was measured.
구체적으로, 부비동 조직 슬라이드를 제작한 후 H&E 염색을 시행하여 상피층 비후 및 염증세포의 유입 등 부비동염 정도를 조직학적으로 확인하였고, 부비동 조직 슬라이드에 시리우스 레드 염색(Sirius Red staining)을 시행하여 호산구(eosinophil) 유입을 확인하였으며, 산성화 톨루이딘 블루 염색(acidified toluidine blue staining)을 통해 비만세포(mast cell)의 유입을 확인하였다.Specifically, after preparing sinus tissue slides, H&E staining was performed to histologically confirm the degree of sinusitis such as epithelial thickening and influx of inflammatory cells. ) was confirmed, and the influx of mast cells was confirmed through acidified toluidine blue staining.
또한, 악성 호산구성 염증 억제 여부를 확인하기 위하여 악성 호산구성 염증 유도에 중요한 PGD2 생성효소(Hepatopoietic prostaglandin D synthase; H-PGDS)의 발현을 면역조직학적 염색으로 확인하였다.In addition, in order to confirm whether malignant eosinophilic inflammation was inhibited, the expression of PGD2 synthase (Hepatopoietic prostaglandin D synthase; H-PGDS), which is important for inducing malignant eosinophilic inflammation, was confirmed by immunohistologic staining.
구체적으로, 부비동 내 PGD2 생성효소 발현 확인을 위해 부비동 조직 슬라이드를 시리우스 레드 염색으로 호산구를 염색하고 항-H-PGDS 항체 (1:100, Cayman Chemicals)로 1시간동안 면역 염색을 진행하였다. 이후, Vectastain ABC 키트 (peroxidase, rabbit IgG) (Vector Labortories, Burlingame, Calif) 및 DAB substrate 키트 (Roche Diagnostics, Mannheim, Germany)를 이용하여 발색시키고 H-PGDS의 발현양을 ImageJ software (National Institures of Health, Bethesda, MD)를 이용하여 분석하였다.Specifically, to confirm the expression of PGD2 synthase in the sinuses, sinus tissue slides were stained with Sirius red eosinophils and immunostained with anti-H-PGDS antibody (1:100, Cayman Chemicals) for 1 hour. Then, the color was developed using the Vectastain ABC kit (peroxidase, rabbit IgG) (Vector Labortories, Burlingame, Calif) and the DAB substrate kit (Roche Diagnostics, Mannheim, Germany), and the expression level of H-PGDS was measured with ImageJ software (National Institutes of Health) , Bethesda, MD).
실시예 2-2: 진세노사이드 F1의 투여 방법에 따른 치료 유효성 평가Example 2-2: Evaluation of therapeutic efficacy according to the administration method of ginsenoside F1
실시예 1의 방법으로 마우스에 만성 부비동염을 유도함과 동시에 진세노사이드 F1을 복강 내로 50mg/kg 이하의 용량 또는 비강 내로 3.5mg/kg 이하의 용량으로 주 3회로 5주간 투여하였다.At the same time as inducing chronic sinusitis in mice by the method of Example 1, ginsenoside F1 was administered intraperitoneally at a dose of 50 mg/kg or less or intranasally at a dose of 3.5 mg/kg or less three times a week for 5 weeks.
그 결과, 복강 내 투여(i.p.) 및 비강 내 투여(i.n.)의 경우 모두 부비동 조직 비후 및 염증세포인 호산구 및 비만세포 의 유입이 감소하였으나, 복강 내 투여에 비해 비강 내 투여의 경우 더욱 감소함을 확인하였다. 이를 통해, 복강 내 투여보다는 비강 내 투여가 조직 내 염증 소견과 면역세포의 침윤을 더 효과적으로 감소시킴으로써 만성 부비동염 치료 효과가 더 우수함을 확인하였다(도 2).As a result, in both intraperitoneal administration (ip) and intranasal administration (in), sinus tissue thickening and influx of eosinophils and mast cells, which are inflammatory cells, were decreased, but it was further reduced in the case of intranasal administration compared to intraperitoneal administration. Confirmed. Through this, it was confirmed that the intranasal administration rather than the intraperitoneal administration more effectively reduced the inflammatory findings and immune cell infiltration in the tissue, thereby confirming that the chronic sinusitis treatment effect was superior (FIG. 2).
실시예 2-3: 일반적인 염증 억제제와 진세노사이드 F1의 만성 부비동염 치료 유효성 비교Example 2-3: Comparison of the treatment efficacy of chronic sinusitis with a general anti-inflammatory inhibitor and ginsenoside F1
상기 만성 부비동염 동물모델에서 진세노사이드 F1을 비강 내로 투여한 경우와 일반적으로 사용되는 염증 억제제를 비강 내 투여한 경우의 효과를 비교하였다. 상기 염증 억제제로는 임상적으로 사용되는 스테로이드성 소염제인 덱사메타손(dexamethasone)을 양성 대조군으로 사용하였다.In the chronic sinusitis animal model, the effects of intranasal administration of ginsenoside F1 and the intranasal administration of a commonly used anti-inflammatory agent were compared. As the inhibitor of inflammation, dexamethasone, a clinically used steroidal anti-inflammatory drug, was used as a positive control.
구체적으로, 실시예 2-1과 동일한 주기로 주 3회 5주간 진세노사이드 F1 및 덱사메타손을 각각 투여하였으며, 진세노사이드 F1을 3.5mg/kg 이하의 용량으로 비강 내 투여한 경우와 덱사메타손을 2mg/kg 이하의 용량으로 비강 내 투여한 경우를 비교하였다.Specifically, ginsenoside F1 and dexamethasone were each administered 3 times a week for 5 weeks at the same cycle as in Example 2-1, and when ginsenoside F1 was intranasally administered at a dose of 3.5 mg/kg or less, and
조직 내 염증 소견과 면역세포인 호산구 및 비만세포의 침윤 정도를 측정한 결과, 만성 부비동염 마우스의 부비동 조직에서 증가한 반면, 진세노사이드 F1을 투여한 경우, 덱사메타손과 마찬가지로 그 정도가 감소함을 확인하였다(도 3).As a result of measuring the inflammatory findings and the infiltration of immune cells, eosinophils and mast cells, in the tissues, it was confirmed that while it increased in the sinus tissues of mice with chronic sinusitis, when ginsenoside F1 was administered, the degree decreased, as with dexamethasone. (Fig. 3).
마우스의 부비동 조직에서 악성 호산구성 염증 유도에 중요한 PGD2 생성효소(H-PGDS)의 발현을 면역조직학적 염색으로 확인한 결과, 만성 부비동염 마우스의 부비동 조직에서 PGD2 생성효소의 발현이 증가한 반면, 진세노사이드 F1 및 덱사메타손 투여에 의해 PGD2 생성효소의 발현이 유의하게 감소됨을 관찰하였다(도 4). Immunohistological staining confirmed the expression of PGD2 synthase (H-PGDS), which is important for induction of malignant eosinophilic inflammation, in the sinus tissues of mice. As a result, the expression of PGD2 synthase was increased in the sinus tissues of mice with chronic sinusitis, whereas ginsenoside It was observed that the expression of PGD2 synthase was significantly reduced by administration of F1 and dexamethasone (FIG. 4).
따라서, 진세노사이드 F1을 투여할 경우 덱사메타손과 마찬가지로 악성 호산구성 염증을 억제함으로써 진세노사이드 F1은 덱사메타손에 비견하는 유의한 치료효과를 보임을 확인하였다.Therefore, when ginsenoside F1 was administered, it was confirmed that ginsenoside F1 showed a significant therapeutic effect comparable to that of dexamethasone by suppressing malignant eosinophilic inflammation like dexamethasone.
실시예 2-4: 진세노사이드 F1의 투여에 의한 염증 사이토카인 발현 변화 측정Example 2-4: Measurement of inflammatory cytokine expression change by administration of ginsenoside F1
진세노사이드 F1 및 덱사메타손의 만성 부비동염 치료 결과를 확인하기 위해 다양한 염증성 사이토카인의 발현 변화를 분석하였다.To confirm the treatment results of chronic sinusitis with ginsenoside F1 and dexamethasone, expression changes of various inflammatory cytokines were analyzed.
구체적으로, 진세노사이드 F1 및 덱사메타손을 각각 투여한 만성 부비동염 마우스의 부비동염 조직에서 mRNA 샘플을 트리졸(Trizol) 시약으로 추출한 후 ReverTraAce qPCR 키트(TOYOBO, Japan)를 이용하여 cDNA로 합성하였다. 이후, 프라이머 및 SYBR Green real time PCR master MIX(TOYOBO)를 함께 혼합하여 Roche light cycler 480(Roche)로 real time PCR을 수행하였다. PCR의 반응 조건으로 먼저 95℃에서 15분동안 반응한 뒤에 95℃에서 30초의 변성, 60℃에서 30초의 어닐링 및 72℃에서 30초의 신장 과정을 45회 반복하였다. 각 유전자의 발현량은 GAPDH의 발현량으로 평준화한 후, 대조군의 발현량과 비교하여 계산하였다. 실험에 사용한 각각의 유전자에 해당하는 프라이머 서열은 하기 표 1과 같다.Specifically, mRNA samples were extracted with Trizol reagent from the sinusitis tissues of chronic sinusitis mice administered with ginsenoside F1 and dexamethasone, respectively, and were synthesized as cDNA using the ReverTraAce qPCR kit (TOYOBO, Japan). Then, the primer and SYBR Green real time PCR master MIX (TOYOBO) were mixed together and real time PCR was performed with a Roche light cycler 480 (Roche). As the reaction conditions of PCR, first reacted at 95°C for 15 minutes, then denaturation at 95°C for 30 seconds, annealing at 60°C for 30 seconds, and elongation at 72°C for 30 seconds were repeated 45 times. The expression level of each gene was calculated by comparing it with the expression level of the control group after normalizing it with the expression level of GAPDH. The primer sequences corresponding to each gene used in the experiment are shown in Table 1 below.
분석 결과, 만성 부비동염 마우스의 부비동 조직에서 IL-4, IL-5, IL-13, 및 IL-1β의 발현이 크게 증가하며, 진세노사이드 F1 및 덱사메타손을 각각 투여한 경우에 알러지성 염증과 관련된 IL-4 및 IL-13의 발현이 공통적으로 감소하였다.As a result of the analysis, the expression of IL-4, IL-5, IL-13, and IL-1β was significantly increased in the sinus tissue of mice with chronic sinusitis, and when ginsenoside F1 and dexamethasone were administered, it was associated with allergic inflammation. The expression of IL-4 and IL-13 was decreased in common.
그러나, 진세노사이드 F1를 투여한 경우 염증 유도와 연관된 IL-1β의 발현은 변화가 없는 반면, 덱사메타손을 투여한 경우에는 감소하였다. 또한, 자연살해세포(Natural killer cell; NK cell)에서 많이 발현되는 것으로 알려진 IFN-γ의 경우 진세노사이드 F1을 투여한 경우에는 유의하게 증가되나, 덱사메타손을 투여한 경우에는 변화가 없었다(도 5).However, when ginsenoside F1 was administered, the expression of IL-1β associated with the induction of inflammation did not change, whereas when dexamethasone was administered, it decreased. In addition, IFN-γ, which is known to be highly expressed in natural killer cells (NK cells), was significantly increased when ginsenoside F1 was administered, but there was no change when dexamethasone was administered (Fig. 5). ).
이를 통해, 진세노사이드 F1은 덱사메타손과 전혀 다른 경로로 만성 부비동염을 치료할 수 있음을 확인하였으며, 염증 억제제와 같은 일반적인 만성 부비동염 치료제에 의해 치료가 불가능하거나 재발되는 난치성 부비동염을 치료할 수 있음을 확인하였다.Through this, it was confirmed that ginsenoside F1 can treat chronic sinusitis by a completely different route than dexamethasone, and it was confirmed that it can treat intractable sinusitis that cannot be treated by general chronic sinusitis drugs such as inflammation inhibitors or that recurs.
실시예 2-5: 진세노사이드 F1이 자연살해세포에 미치는 영향 평가Example 2-5: Evaluation of the effect of ginsenoside F1 on natural killer cells
만성 부비동염에서 진세노사이드 F1이 자연살해세포에 미치는 영향을 평가하기 위하여, 만성 부비동염 동물모델에서 자연살해세포 특이적 항체를 투여하여 자연살해세포가 결핍된 마우스 모델을 확립하였다.In order to evaluate the effect of ginsenoside F1 on NK cells in chronic sinusitis, a NK cell-deficient mouse model was established by administering a NK cell-specific antibody in an animal model of chronic sinusitis.
구체적으로, 실시예 1에서 제작한 호산구성 만성 부비동염 마우스 모델에 일반적으로 자연살해세포를 결핍시키기 위해 사용되는 항-ASGM1 항체(anti-asialo GM1 antibody; anti-ASGM1 antibody, Cedarlane)를 10㎕의 용량으로 5일 간격으로 7회 복강 내 투여(i.p.)함으로써, 상기 마우스 모델에서 자연살해세포를 결핍시켰다(도 6).Specifically, in the mouse model of chronic eosinophilic sinusitis prepared in Example 1, anti-ASGM1 antibody (anti-asialo GM1 antibody; anti-ASGM1 antibody, Cedarlane) generally used to deficient natural killer cells in a dose of 10 μl By intraperitoneal administration (ip) 7 times at intervals of 5 days, the mouse model was depleted of natural killer cells (FIG. 6).
자연살해세포의 결핍 여부를 확인하기 위하여, 항-ASGM1 항체가 처리된 마우스의 비장(spleen)을 분리하였다. 분리된 비장세포를 형광이 표지된 항-CD3ε 항체 및 항-NKp46 항체로 염색하고 유세포 분석(flow cytometry)을 이용하여 FSC(forward-scattered light)와 SSC(side-scattered light)를 산출함으로써 일차적으로 림프구 세포를 확인하였으며, 이 중 자연살해세포(CD3- NKp46+ 집단)의 분포를 측정하였다.In order to determine whether natural killer cells are deficient, the spleen of mice treated with anti-ASGM1 antibody was isolated. The isolated splenocytes were stained with fluorescence-labeled anti-CD3ε antibody and anti-NKp46 antibody, and flow cytometry was used to calculate forward-scattered light (FSC) and side-scattered light (SSC). Lymphocyte cells were identified, and the distribution of natural killer cells (CD3- NKp46+ population) was measured.
그 결과, 항-ASGM1 항체 투여로 자연살해세포가 결핍되었음을 확인하였다(도 7).As a result, it was confirmed that natural killer cells were deficient by administration of the anti-ASGM1 antibody (FIG. 7).
이후, 상기 자연살해세포 결핍 만성 부비동염 동물모델에 진세노사이드 F1 및 덱사메타손을 각각 투여한 후 만성 부비동염의 치료 효과를 비교하기 위하여 상피층 비후 정도, 점막 두께, 및 염증세포의 유입을 측정한 결과, 진세노사이드 F1에 의한 치료 효과는 자연살해세포 결핍에 의해 유의하게 감소한 반면, 덱사메타손에 의한 치료 효과는 자연살해세포 결핍에 큰 영향을 받지 않음을 확인하였다(도 8).Thereafter, in order to compare the therapeutic effects of chronic sinusitis after administration of ginsenoside F1 and dexamethasone to the natural killer cell-deficient chronic sinusitis animal model, respectively, the degree of epithelial thickening, mucosal thickness, and influx of inflammatory cells were measured. While the therapeutic effect of senoside F1 was significantly reduced by the NK cell deficiency, it was confirmed that the therapeutic effect of dexamethasone was not significantly affected by the NK cell deficiency (FIG. 8).
이를 통해, 진세노사이드 F1은 덱사메타손과 달리 자연살해세포를 매개로 하여 만성 부비동염을 치료하는 것임을 확인하였다. 따라서, 일반적인 염증 억제제 또는 항생제와는 다른 기전을 갖는 것으로서, 일반적인 염증 억제제 또는 항생제를 투여하여도 치료 효과가 나타나지 않았거나 재발하는 난치성 부비동염 또는 호산성 만성 부비동염 치료에 진세노사이드 F1이 효과적임을 확인하였다.Through this, it was confirmed that ginsenoside F1 treats chronic sinusitis by mediated natural killer cells, unlike dexamethasone. Therefore, it was confirmed that ginsenoside F1 is effective in treating intractable sinusitis or chronic eosinophilic sinusitis that does not show any therapeutic effect even after administration of a general anti-inflammatory or antibiotic, as it has a mechanism different from that of general anti-inflammatory or antibiotic. .
이상의 설명으로부터, 본 발명이 속하는 기술분야의 당업자는 본 발명이 그 기술적 사상이나 필수적 특징을 변경하지 않고서 다른 구체적인 형태로 실시될 수 있다는 것을 이해할 수 있을 것이다. 이와 관련하여, 이상에서 기술한 실시예들은 모든 면에서 예시적인 것이며 한정적인 것이 아닌 것으로 이해해야만 한다. 본 발명의 범위는 상기 상세한 설명보다는 후술하는 특허 청구범위의 의미 및 범위 그리고 그 등가 개념으로부터 도출되는 모든 변경 또는 변형된 형태가 본 발명의 범위에 포함되는 것으로 해석되어야 한다.From the above description, those skilled in the art to which the present invention pertains will understand that the present invention may be embodied in other specific forms without changing the technical spirit or essential characteristics thereof. In this regard, it should be understood that the embodiments described above are illustrative in all respects and not restrictive. The scope of the present invention should be construed as being included in the scope of the present invention, rather than the above detailed description, all changes or modifications derived from the meaning and scope of the claims to be described later and their equivalents.
<110> Intelligent Synthetic Biology Center <120> Composition for preventing or treating chronic rhinosinusitis comprising Ginsenoside F1 <130> KPA200317-KR <160> 14 <170> KoPatentIn 3.0 <210> 1 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> Forward <400> 1 acagcaaggc gaaaaaggat g 21 <210> 2 <211> 19 <212> DNA <213> Artificial Sequence <220> <223> Reverse <400> 2 tggtggacca ctcggatga 19 <210> 3 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> Forward <400> 3 catcggcatt ttgaacgagg t 21 <210> 4 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> Reverse <400> 4 catgcagtag acatggcaga a 21 <210> 5 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> Forward <400> 5 gcaatgagac gatgaggctt c 21 <210> 6 <211> 23 <212> DNA <213> Artificial Sequence <220> <223> Reverse <400> 6 gcccctgaaa gatttctcca atg 23 <210> 7 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Forward <400> 7 agaccagact cccctgtgca 20 <210> 8 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> Reverse <400> 8 tgggtcctgt agatggcatt g 21 <210> 9 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> Forward <400> 9 gaatgacctg ttctttgaag t 21 <210> 10 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> Reverse <400> 10 tttgttgttc atctcgaagc c 21 <210> 11 <211> 19 <212> DNA <213> Artificial Sequence <220> <223> Forward <400> 11 cgcaaaagtg agctccaga 19 <210> 12 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Reverse <400> 12 tgagcttccc agatcacaga 20 <210> 13 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> Forward <400> 13 agtatgatga catcaagaag g 21 <210> 14 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> Reverse <400> 14 atggtattca agagagtagg g 21 <110> Intelligent Synthetic Biology Center <120> Composition for preventing or treating chronic rhinosinusitis comprising Ginsenoside F1 <130> KPA200317-KR <160> 14 <170> KoPatentIn 3.0 <210> 1 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> <400> 1 acagcaaggc gaaaaaggat g 21 <210> 2 <211> 19 <212> DNA <213> Artificial Sequence <220> <223> Reverse <400> 2 tggtggacca ctcggatga 19 <210> 3 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> <400> 3 catcggcatt ttgaacgagg t 21 <210> 4 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> Reverse <400> 4 catgcagtag acatggcaga a 21 <210> 5 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> <400> 5 gcaatgagac gatgaggctt c 21 <210> 6 <211> 23 <212> DNA <213> Artificial Sequence <220> <223> Reverse <400> 6 gcccctgaaa gatttctcca atg 23 <210> 7 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> <400> 7 agaccagact cccctgtgca 20 <210> 8 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> Reverse <400> 8 tgggtcctgt agatggcatt g 21 <210> 9 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> <400> 9 gaatgacctg ttctttgaag t 21 <210> 10 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> Reverse <400> 10 tttgttgttc atctcgaagc c 21 <210> 11 <211> 19 <212> DNA <213> Artificial Sequence <220> <223> <400> 11 cgcaaaagtg agctccaga 19 <210> 12 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Reverse <400> 12 tgagcttccc agatcacaga 20 <210> 13 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> <400> 13 agtatgatga catcaagaag g 21 <210> 14 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> Reverse <400> 14 atggtattca agagagtagg g 21
Claims (9)
A pharmaceutical composition for preventing or treating chronic rhinosinusitis (CRS) comprising ginsenoside F1 as an active ingredient.
The pharmaceutical composition of claim 1, wherein the chronic sinusitis is eosinophilic chronic sinusitis.
The pharmaceutical composition of claim 1, wherein the chronic sinusitis is intractable sinusitis.
The pharmaceutical composition according to claim 3, wherein the refractory sinusitis cannot be treated with an inflammatory inhibitor or antibiotic or recurs.
The pharmaceutical composition according to claim 1, wherein the ginsenoside F1 promotes the activity of natural killer cells (NK cells).
The pharmaceutical composition of claim 1, wherein the ginsenoside F1 increases the expression of cytokine IFN-γ.
A food composition for preventing or improving chronic sinusitis comprising ginsenoside F1 as an active ingredient.
A feed composition for preventing or improving chronic sinusitis comprising ginsenoside F1 as an active ingredient.
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