KR20090131417A - Compositions for preventing or treating allergy diseases - Google Patents
Compositions for preventing or treating allergy diseases Download PDFInfo
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- KR20090131417A KR20090131417A KR1020080057273A KR20080057273A KR20090131417A KR 20090131417 A KR20090131417 A KR 20090131417A KR 1020080057273 A KR1020080057273 A KR 1020080057273A KR 20080057273 A KR20080057273 A KR 20080057273A KR 20090131417 A KR20090131417 A KR 20090131417A
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Abstract
Description
본 발명은 홍삼 추출물, 자소 추출물, 알로에 추출물, 어성초 추출물 및 국화 추출물을 유효성분으로 포함하는 알레르기(allergy) 질환 예방 또는 치료용 조성물에 관한 것이다.The present invention relates to a composition for preventing or treating allergy diseases, including red ginseng extract, shiso extract, aloe extract, eochocho extract and chrysanthemum extract as active ingredients.
최근 들어, 사회 환경과 식생활의 변화로 저하로 알레르기 환자가 급증하고 있다. In recent years, allergic patients are rapidly increasing due to changes in the social environment and diet.
알레르기(Allergy) 반응은 무해한 항원, 즉 알레르겐(Allergen)에 반응하여 생성된 IgE 항체를 지닌 개인이 계속하여 동일 알레르겐에 노출되었을 때 일어난다. 알레르겐은 이미 노출된 조직에 있는 IgE 결합 비만세포를 활성화시켜, 알레르기(allergy)의 특징적인 반응들을 연속적으로 일어나게 한다. 알레르기는 과민반응들(Hypersensitivity reactions)이라 칭하는 면역계 반응들의 여러 가지 분류들 중의 하나이다. 이때, 면역계 반응들이란 조직손상을 일으키며 심각한 질병을 일으킬 수 있는 유해한 면역반응들을 말한다.Allergy reactions occur when individuals with IgE antibodies produced in response to harmless antigens, ie allergens, continue to be exposed to the same allergens. Allergens activate IgE-binding mast cells in already exposed tissues, causing a series of characteristic reactions of allergy. Allergies are one of several classes of immune system reactions called hypersensitivity reactions. In this case, the immune system reactions are harmful immune reactions that can cause tissue damage and serious disease.
과민반응은 Coombs와 Gell에 의하여 네 가지형으로 분류되는데, 알레르기는 종종 Type Ⅰ 과민반응과 동일시 사용된다. 서구인의 40% 정도는 흔히 접할 수 있는 다양한 종류의 공통 환경 알레르겐들에 의해 점차 상승하는 방식으로 IgE 반응이 악화되는 경향이 있다. 이러한 상태를 아토피(atopy)라고 부르며 여러 유전적인 영향을 받는 것으로 알려져 있다. 아직도 많은 사람들은 이러한 극소량의 알레르겐에 자극되고 심지어 생명을 위협하는 IgE 항체반응이 일어난다. 대표적인 질병으로는 알레르기 비염, 천식 또는 아나필락시스(과민증)이 있다.Hypersensitivity is classified into four types by Coombs and Gell. Allergies are often used to identify Type I hypersensitivity. About 40% of Westerners tend to exacerbate the IgE response in a way that is gradually elevated by various common environmental allergens. This condition is called atopy and is known to have several genetic effects. Still, many people are stimulated by these tiny amounts of allergens and even life-threatening IgE antibody responses occur. Representative diseases include allergic rhinitis, asthma or anaphylaxis.
현재, 알레르기 질환의 치료에는 항히스타민제나 스테로이드제 등이 많이 사용되고 있다. 그러나 이러한 약물은 그 효과가 일시적인 경우가 대부분이고 부작용이 심한 경우도 많다. 따라서 알레르기 질환의 예방 및 개선효과를 가지면서 부작용이 적고 효과가 지속적인 새로운 물질의 개발이 필요하다.Currently, antihistamines, steroids, and the like are frequently used for the treatment of allergic diseases. However, these drugs are often temporary and often have severe side effects. Therefore, there is a need for the development of new substances that have the effect of preventing and improving allergic diseases and having fewer side effects and lasting effects.
본 명세서 전체에 걸쳐 다수의 특허문헌이 참조되고 그 인용이 표시되어 있다. 인용된 특허문헌의 개시 내용은 그 전체로서 본 명세서에 참조로 삽입되어 본 발명이 속하는 기술 분야의 수준 및 본 발명의 내용이 보다 명확하게 설명된다.Throughout this specification many patent documents are referenced and their citations are indicated. The disclosures of the cited patent documents are incorporated by reference herein in their entirety, and the level of the technical field to which the present invention belongs and the contents of the present invention are more clearly described.
본 발명자들은 홍삼 및 천연 생약재를 소재로 하여 알레르기 질환을 예방 또는 치료할 수 있는 물질을 개발하고자 노력하였다. 그 결과, 홍삼 추출물, 자소추출물, 알로에 추출물, 어성초 추출물 및 국화 추출물을 혼합하여 제조한 조성물을 투여하면, 면역 과민 반응을 유발한 모델에서 히스타민 유리 억제, IgE 생성 억제, 림프구 증식 및 사이토카인의 분비(특히, IL-2 및 IFN-γ)에 우수한 상승적(synergic) 효과를 가짐을 확인함으로써, 본 발명을 완성하게 되었다. The present inventors have tried to develop a material that can prevent or treat allergic diseases based on red ginseng and natural herbal medicines. As a result, administration of a composition prepared by mixing red ginseng extract, shoot extract, aloe extract, echochocho extract and chrysanthemum extract, inhibited histamine release, IgE production, lymphocyte proliferation and cytokine secretion in a model that caused immune hypersensitivity reactions. The present invention has been completed by confirming that it has an excellent synergic effect on IL-2 and IFN-γ.
따라서, 본 발명의 목적은 알레르기 질환의 예방 또는 치료용 조성물을 제공하는 데 있다.Accordingly, it is an object of the present invention to provide a composition for preventing or treating allergic diseases.
본 발명의 다른 목적 및 이점은 하기의 발명의 상세한 설명, 청구범위 및 도면에 의해 보다 명확하게 된다.Other objects and advantages of the present invention will become apparent from the following detailed description, claims and drawings.
본 발명의 일 양태에 따르면, 본 발명은 홍삼 추출물, 자소 추출물, 알로에 추출물, 어성초 추출물 및 국화 추출물을 유효성분으로 포함하는 알레르기(allergy) 질환 예방 또는 치료용 조성물을 제공한다.According to an aspect of the present invention, the present invention provides a composition for preventing or treating allergy disease, which comprises red ginseng extract, jaso extract, aloe extract, Echochocho extract and chrysanthemum extract as active ingredients.
본 발명자들은 홍삼 및 천연 생약재를 소재로 하여 알레르기 질환을 예방 또 는 치료할 수 있는 물질을 개발하고자 노력하였다. 그 결과, 홍삼 추출물, 자소추출물, 알로에 추출물, 어성초 추출물 및 국화 추출물을 혼합하여 제조한 조성물을 투여하면, 면역 과민 반응을 유발한 모델에서 히스타민 유리 억제, IgE 생성 억제, 림프구 증식 및 사이토카인의 분비(특히, IL-2 및 IFN-γ)에 우수한 상승적(synergic) 효과를 가짐을 확인하였다.The present inventors tried to develop a material that can prevent or treat allergic diseases based on red ginseng and natural herbal medicines. As a result, administration of a composition prepared by mixing red ginseng extract, shoot extract, aloe extract, echochocho extract and chrysanthemum extract, inhibited histamine release, IgE production, lymphocyte proliferation and cytokine secretion in a model that caused immune hypersensitivity reactions. (In particular, IL-2 and IFN-γ) was confirmed to have a good synergic effect.
본 발명에서 유효성분으로 이용되는 “홍삼(Panax ginseng C.A. Meyer)”은 증기 또는 태양-건조, 바람직하게는 증기를 통하여 인삼을 가열하여 제조된 것을 의미한다. 기재의 편의상, 본 발명은 인삼 또는 홍삼에 적용되는 것으로 기재되어 있다.Red ginseng used as an active ingredient in the present invention ( Panax ginseng CA Meyer) ”means prepared by heating ginseng through steam or sun-drying, preferably steam. For convenience of description, the present invention is described as being applied to ginseng or red ginseng.
홍삼(Panax ginseng C.A. Meyer)은 외적 유해인자에 대한 비특이적인 생체 저항력을 증진시켜 주며 물리적, 화학적 또는 생물학적인 외적 변화에 대해 생체를 정상화시켜 주는 “범적응적 활성(adaptogen)”으로 작용하여 면역기능을 조절해 주는 기능이 있으며, 한편, 홍삼의 주요 효능 성분인 사포닌성분은 면역인자 사멸을 억제시키는 효능이 있다 (Choi DH et al., Inhibitory effects of ginseng saponin fractions on dexamethasone-induced thymus apoptosis. Korean J. Ginseng S챠. 21(3): 160-168, (1997)).Red Ginseng ( Panax ginseng CA Meyer) enhances nonspecific bio-resistance against external harmful factors and regulates immune function by acting as “adaptogens” that normalize the living body against physical, chemical or biological external changes. On the other hand, saponin, the main potent ingredient of red ginseng, has the effect of suppressing immune factor death (Choi DH et al., Inhibitory effects of ginseng saponin fractions on dexamethasone-induced thymus apoptosis.Korean J. Ginseng S 21 (3): 160-168, (1997).
본 명세서에서 용어 “인삼”은 고려삼(Panax ginseng), 회기삼(P. quiquefolius), 전칠삼(P. notoginseng), 죽절삼(P. japonicus), 삼엽삼(P. trifolium), 히말라야삼(P. pseudoginseng), 베트남삼(P. vietnamensis) 및 미국삼 (Panax quinquefolium)을 포함하나, 이에 한정되지 않는다. 바람직하게는, 본 발명에서 이용되는 인삼은 고려삼(Panax ginseng)이다.As used herein, the term “ginseng” refers to Korean ginseng ( Panax ginseng), three sessions (P. quiquefolius), jeonchilsam (P. notoginseng), jukjeol three (P. japonicus), three yeopsam (P. trifolium), Himalayan cedar (P. pseudoginseng), Vietnam, three (P. vietnamensis) and American hemp ( Panax quinquefolium ), but is not limited thereto. Preferably, the ginseng used in the present invention is Korean ginseng ( Panax ginseng ).
본 발명에서 유효성분으로 이용되는 “자소(紫蘇, 학명: Perilla frutescens Britton var . crispa (Thunb.) Decne ., 영명: acute common perilla 또는 purple common perilla)”는 꿀풀과(Labiatae)에 속하는 1년생 초본으로, 한방에서는 해열, 발한, 거담, 건위, 행기, 해독 또는 안태 등의 효능이 있다고 하여, 감기, 오한발열, 해소, 오심, 구토, 소화불량, 어중독 또는 태동불안 등에 사용되어 왔다.Used in the present invention as the active ingredient "Jangja (紫蘇, Scientific name: Perilla frutescens Britton var . crispa (Thunb.) Decne ., English: acute common perilla or purple common perilla ”is a herbaceous herb that belongs to the labiatae. It has been used for colds, chills, fever, nausea, vomiting, indigestion, fish poisoning or infertility.
본 발명에서 유효성분으로 이용되는 “알로에”는 전통적으로 인체의 면역기능을 회복하거나 유지하는 효능을 갖고 있으며, 알로에에 함유되어 있는 알프로젠이라는 성분은 알러지 반응을 효과적으로 억제하는 것으로 알려져 있다."Aloe" used as an active ingredient in the present invention traditionally has the effect of restoring or maintaining the immune function of the human body, and an ingredient called alprogen contained in aloe is known to effectively suppress allergic reactions.
본 발명에서 유효성분으로 이용되는 “어성초(魚腥草)”는 항균작용, 인플루엔자바이러스 등에 대한 항바이러스작용, 모세혈관 강화작용, 혈관 확장작용 또는 이뇨작용 등이 있는 것으로 알려져 있다."Eoseongcho" used as an active ingredient in the present invention is known to have antibacterial action, antiviral action against influenza virus, capillary potentiation effect, vasodilation effect or diuretic effect.
본 발명에서 유효성분으로 이용되는 “국화”는 한방에서 해열, 해독, 진통 또는 소염약 등으로서 감기에 의한 발열 두통 등에 사용되며, 또한 최근에는 혈압강하(血壓降下) 작용이 있다는 것이 밝혀져서 동맥경화증 및 고콜레스테롤혈증 등에도 이용되고 있다."Chrysanthemum" used as an active ingredient in the present invention is used for fever headache caused by cold as antipyretic, detoxification, analgesic or anti-inflammatory medicine in oriental medicine, and also recently, it has been found that there is a blood pressure lowering effect (血壓 降下) action atherosclerosis And hypercholesterolemia.
본 발명의 유효성분으로 이용되는 홍삼, 자소, 알로에, 어성초 또는 국화 추출물은 당업계에 공지된 통상적인 추출 과정에서 의해 얻을 수 있는 추출물을 포함 한다. 예를 들어, (a) 물, (b) 탄소수 1-4의 무수 또는 함수 저급 알코올 (메탄올, 에탄올, 프로판올, 부탄올 등), (c) 상기 저급 알코올과 물과의 혼합용매, (d) 아세톤, (e) 에틸 아세테이트, (f) 클로로포름, (g) 부틸아세테이트 또는 (h) 1,3-부틸렌글리콜을 추출용매로 하여 얻은 추출물이 본 발명의 유효성분에 포함된다. 바람직하게는, 본 발명의 추출물은 물, 에탄올 또는 에탄올과 물의 혼합용매를 추출용매로 하여 얻은 것이며, 가장 바람직하게는 물을 추출용매로 하여 얻은 것이다.Red ginseng, shiso, aloe, fish vinegar or chrysanthemum extract used as an active ingredient of the present invention includes extracts obtained by conventional extraction procedures known in the art. For example, (a) water, (b) anhydrous or hydrous lower alcohol having 1 to 4 carbon atoms (methanol, ethanol, propanol, butanol, etc.), (c) a mixed solvent of the lower alcohol with water, (d) acetone The extract obtained by using (e) ethyl acetate, (f) chloroform, (g) butyl acetate or (h) 1,3-butylene glycol as an extraction solvent is included in the active ingredient of the present invention. Preferably, the extract of the present invention is obtained by using water, ethanol or a mixed solvent of ethanol and water as the extraction solvent, and most preferably obtained by using water as the extraction solvent.
한편, 본 발명의 추출물은 상기한 추출 용매뿐만 아니라, 다른 추출 용매를 이용하여도 실질적으로 동일한 효과를 나타내는 본 발명의 추출물이 얻어질 수 있다는 것은 당업자에게 자명한 것이다. 또한, 본 발명의 추출물은 상술한 추출 용매에 의한 추출물뿐만 아니라, 통상적인 정제 과정을 거친 추출물도 포함한다. 예컨대, 일정한 분자량 컷-오프 값을 갖는 한외여과막을 이용한 분리, 다양한 크로마토그래피 (크기, 전하, 소수성 또는 친화성에 따른 분리를 위해 제작된 것)에 의한 분리 등, 추가적으로 실시된 다양한 정제 방법을 통해 얻어진 분획도 본 발명의 추출물에 포함되는 것이다.On the other hand, it will be apparent to those skilled in the art that the extract of the present invention can be obtained in addition to the above-described extraction solvent, as well as the extract of the present invention exhibiting substantially the same effects using other extraction solvents. In addition, the extract of the present invention includes not only the extract by the above-described extraction solvent, but also the extract that has undergone a conventional purification process. Obtained by various additional purification methods, such as, for example, separation using ultrafiltration membranes having a constant molecular weight cut-off value, separation by various chromatography (manufactured for separation according to size, charge, hydrophobicity or affinity). The fraction is also included in the extract of the present invention.
본 발명의 추출물은 감압 증류 및 동결 건조 또는 분무 건조 등과 같은 추가적인 과정에 의해 분말 상태로 제조될 수 있다.Extracts of the present invention may be prepared in powder form by additional processes such as distillation under reduced pressure and freeze drying or spray drying.
한편, 본 발명에서 이용되는 인삼 또는 홍삼 추출물은 인삼 또는 홍삼 추출물을 장내 세균 또는 유산균을 이용하여 발효한 발효 홍삼액도 포함한다.On the other hand, ginseng or red ginseng extract used in the present invention also includes fermented red ginseng liquid fermented ginseng or red ginseng extract using intestinal bacteria or lactic acid bacteria.
본 발명의 바람직한 구현예에 따르면, 본 발명의 조성물에 이용되는 유효성분은 조성물 전체 중량에 대하여 0.1-40%를 포함한다. 보다 바람직하게는 10-30%, 가장 바람직하게는 15-25%이다. 유효성분의 중량이 0.1 중량% 미만일 때는 그 효과가 나타나기 어렵고, 40 중량%를 초과하는 경우에는 함량의 증가에 따른 효과의 증가가 매우 미약하고, 제형상의 안정성이 확보되지 않는 문제점이 있다.According to a preferred embodiment of the present invention, the active ingredient used in the composition of the present invention comprises 0.1-40% based on the total weight of the composition. More preferably 10-30%, and most preferably 15-25%. When the weight of the active ingredient is less than 0.1% by weight, the effect is less likely to appear, when it exceeds 40% by weight, the increase in effect due to the increase of the content is very weak, there is a problem that the stability in the formulation is not secured.
본 발명의 다른 바람직한 구현예에 따르면, 본 발명의 조성물에 이용되는 유효성분은 유효성분 전체 중량에 대하여 홍삼 추출물 39-60 중량%, 자소 추출물 0.5-5 중량%, 알로에 추출물 0.1-1 중량%, 어성초 추출물 25-35 중량% 및 국화 추출물 14.4-20 중량%를 포함하고, 보다 바람직하게는 홍삼 추출물 46.6-55 중량%, 자소 추출물 1.0-2.7 중량%, 알로에 추출물 0.18-0.7 중량%, 어성초 추출물 28.2-32 중량% 및 국화 추출물 15.62-18 중량%, 가장 바람직하게는 홍삼 추출물 50-52 중량%, 자소 추출물 1.0-2.0 중량%, 알로에 추출물 0.2-0.5 중량%, 어성초 추출물 29.2-30.5 중량% 및 국화 추출물 16.5-17.0 중량%로 포함되는 것이다.According to another preferred embodiment of the present invention, the active ingredient used in the composition of the present invention is 39-60% by weight, ginseng extract 0.5-5%, aloe extract 0.1-1% by weight, based on the total weight of the active ingredient, 25-35% by weight of Echochocho extract and 14.4-20% by weight of chrysanthemum extract, more preferably 46.6-55% by weight of red ginseng extract, 1.0-2.7% by weight of perilla extract, 0.18-0.7% by weight of aloe extract, 28.2% of Echocho extract -32 wt% and 15.62-18 wt% chrysanthemum extract, most preferably 50-52 wt% red ginseng extract, 1.0-2.0 wt% perilla extract, 0.2-0.5 wt% aloe extract, 29.2-30.5 wt% chrysanthemum extract and chrysanthemum It is included in the extract 16.5-17.0% by weight.
본 명세서에서 용어 “알레르기 질환”은 인체의 어떤 물질에 대한 과민증, 즉 외부로부터 들어온 물질에 대한 신체 면역계의 과도한 반응을 일으켜서 유발되는 다양한 질환, 질병 또는 이상 상태를 의미한다. 특히, 외부로부터 들어온 물질에 의해 히스타민과 같은 매개물질이 유리되어 질병이 유발되는 과민증을 말한다. 본 발명의 조성물이 적용되는 알레르기 질환으로는, 바람직하게는, IgE에 의해 매개되는 알레르기 질환(제 1 형 과민 반응) 또는 항원(혹은 알레르겐)에 감작 된 T세포에 의해 매개되는 알레르기 질환(제 4 항 과민 반응)이고, 보다 바람직하게는 IgE 매개 알레르기 질환은 기관지 천식, 알레르기성 비염, 아토피성 피부염, 알레르기성 결막염, 알레르기성 중이염, 두드러기 및 아나필락시 쇼크(anaphylatic shock)이고, T세포 매개 알레르기 질환은 접촉성 과민증, 알레르기성 접촉성 피부염, 세균 알레르기, 진균 알레르기, 바이러스 알레르기, 약물알레르기, 갑상선염 및 알레르기성 뇌염이며, 가장 바람직하게는 알레르기성 비염, 아토피성 피부염 및 약물알레르기를 포함한다.As used herein, the term “allergic disease” refers to various diseases, diseases or abnormalities caused by hypersensitivity to certain substances in the human body, ie, excessive reaction of the body's immune system to substances from outside. In particular, it refers to hypersensitivity in which a mediator, such as histamine, is released by a substance from the outside to cause disease. As an allergic disease to which the composition of the present invention is applied, preferably, an allergic disease mediated by IgE (type 1 hypersensitivity reaction) or an allergic disease mediated by T cells sensitized to an antigen (or allergen) (fourth) Anti-hypersensitivity reactions), more preferably IgE mediated allergic diseases are bronchial asthma, allergic rhinitis, atopic dermatitis, allergic conjunctivitis, allergic otitis media, urticaria and anaphylatic shock, Contact hypersensitivity, allergic contact dermatitis, bacterial allergy, fungal allergy, viral allergy, drug allergy, thyroiditis and allergic encephalitis, most preferably allergic rhinitis, atopic dermatitis and drug allergy.
본 발명의 가장 큰 특징은, 본 발명의 유효성분은 면역 과민 반응 유발 모델에서 우수한 히스타민 유리 억제 및 IgE 생성 억제 효과를 나타내어 IgE 매개 알레르기 질환의 예방 또는 치료 효과를 가지고, T세포 매개 알레르기 질환을 일으키는 인자인 림프구 및 사이토카인과 관련하여 우수한 림프구 증식, 및 IL-2와 IFN-γ의 증가 효과를 나타내어 알레르기 질환을 예방 또는 치료하는데 효과가 높다. 또한, 홍삼 추출물, 자소 추출물, 알로에 추출물, 어성초 추출물 또는 국화 추출물을 각각 복용하였을 때 보다, 본 발명의 복합 유효성분을 포함하는 조성물을 복용하였을 때 상승적 효과를 가지며, 이러한 알레르기 질환의 예방 또는 치료 효과는 종래 알레르기 질환의 치료제로 이용되고 있는 지르텍 보다 우수하다.The greatest feature of the present invention, the active ingredient of the present invention exhibits excellent histamine free inhibition and IgE production inhibitory effect in the immune hypersensitivity reaction model, has a prophylactic or therapeutic effect of IgE mediated allergic disease, causing T cell mediated allergic disease Excellent lymphocyte proliferation and increased effects of IL-2 and IFN- [gamma] in relation to factors such as lymphocytes and cytokines are highly effective in preventing or treating allergic diseases. In addition, red ginseng extract, shiso extract, aloe extract, Echo extract or chrysanthemum extract, respectively, than when taking the composition containing the composite active ingredient of the present invention has a synergistic effect, and prevent or treat these allergic diseases Is superior to Zirtec, which is conventionally used as a treatment for allergic diseases.
또한, 본 발명의 조성물은 약제학적 또는 식품 조성물로 제조될 수 있다.In addition, the compositions of the present invention may be prepared as pharmaceutical or food compositions.
본 발명의 조성물이 약제학적 조성물로 제조되는 경우, 본 발명의 약제학적 조성물은 약제학적으로 허용되는 담체를 포함한다. 본 발명의 약제학적 조성물에 포함되는 약제학적으로 허용되는 담체는 제제 시에 통상적으로 이용되는 것으로서, 락토스, 덱스트로스, 수크로스, 솔비톨, 만니톨, 전분, 아카시아 고무, 인산 칼슘, 알기네이트, 젤라틴, 규산 칼슘, 미세결정성 셀룰로스, 폴리비닐피롤리돈, 셀룰로스, 물, 시럽, 메틸 셀룰로스, 메틸히드록시벤조에이트, 프로필히드록시벤조에이트, 활석, 스테아르산 마그네슘 및 미네랄 오일 등을 포함하나, 이에 한정되는 것은 아니다. 본 발명의 약제학적 조성물은 상기 성분들 이외에 윤활제, 습윤제, 감미제, 향미제, 유화제, 현탁제, 보존제등을 추가로 포함할 수 있다. 적합한 약제학적으로 허용되는 담체 및 제제는 Remington's Pharmaceutical Sciences (19th ed., 1995)에 상세히 기재되어 있다.When the composition of the present invention is made into a pharmaceutical composition, the pharmaceutical composition of the present invention includes a pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers included in the pharmaceutical compositions of the present invention are those commonly used in the preparation, such as lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia rubber, calcium phosphate, alginate, gelatin, Calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil, and the like It doesn't happen. The pharmaceutical composition of the present invention may further include lubricants, wetting agents, sweeteners, flavoring agents, emulsifiers, suspending agents, preservatives and the like in addition to the above components. Suitable pharmaceutically acceptable carriers and formulations are described in detail in Remington's Pharmaceutical Sciences (19th ed., 1995).
본 발명의 약제학적 조성물은 경구 또는 비경구 투여 (예컨대, 정맥내 투여, 복강내 투여, 근육내 투여, 피하 투여, 또는 국부 투여)할 수 있다.The pharmaceutical compositions of the present invention may be oral or parenteral (eg, intravenous, intraperitoneal, intramuscular, subcutaneous, or topical).
본 발명의 약제학적 조성물의 적합한 투여량은 제제화 방법, 투여 방식, 환자의 연령, 체중, 성, 병적 상태, 음식, 투여 시간, 투여 경로, 배설 속도 및 반응 감응성과 같은 요인들에 의해 다양하게 처방될 수 있다. 한편, 본 발명의 약제학적 조성물의 투여량은 바람직하게는 1일 당 0.01-100 ㎎/kg(체중)이다.Suitable dosages of the pharmaceutical compositions of the present invention may vary depending on factors such as the formulation method, mode of administration, age, weight, sex, morbidity, condition of food, time of administration, route of administration, rate of excretion and response to response of the patient. Can be. On the other hand, the dosage of the pharmaceutical composition of the present invention is preferably 0.01-100 mg / kg body weight per day.
본 발명의 약제학적 조성물은 당해 발명이 속하는 기술분야에서 통상의 지식을 가진 자가 용이하게 실시할 수 있는 방법에 따라, 약제학적으로 허용되는 담체 및/또는 부형제를 이용하여 제제화함으로써 단위 용량 형태로 제조되거나 또는 다용량 용기 내에 내입시켜 제조될 수 있다. 이때 제형은 산제, 과립제, 정제, 캅셀 제, 현탁액, 에멀젼, 시럽, 에어로졸 등의 경구형 제형, 연고, 크림등의 외용제, 좌제 및 멸균 주사용액 등을 비롯하여 약제학적 제제에 적합한 어떠한 형태로든 사용할 수 있으며, 분산제 또는 안정화제를 추가적으로 포함할 수 있다.The pharmaceutical compositions of the present invention may be prepared in unit dose form by formulating with a pharmaceutically acceptable carrier and / or excipient according to methods which can be easily carried out by those skilled in the art. Or may be prepared by incorporation into a multi-dose container. The formulations may be used in any form suitable for pharmaceutical preparations, including powders, granules, tablets, capsules, suspensions, emulsions, syrups, oral formulations such as aerosols, external preparations such as ointments, creams, suppositories, and sterile injectable solutions. It may further include a dispersant or stabilizer.
본 발명의 조성물이 식품 조성물로 제조되는 경우, 상술한 본 발명의 유효성분뿐만 아니라, 식품 제조 시에 통상적으로 첨가되는 성분을 포함하며, 예를 들어, 단백질, 탄수화물, 지방, 영양소, 조미제 및 향미제를 포함한다. 상술한 탄수화물의 예는 모노사카라이드, 예를 들어, 포도당, 과당 등; 디사카라이드, 예를 들어 말토스, 슈크로스, 올리고당 등; 및 폴리사카라이드, 예를 들어 덱스트린, 사이클로덱스트린 등과 같은 통상적인 당 및 자일리톨, 소르비톨, 에리트리톨 등의 당알콜이다. 향미제로서 천연 향미제 [타우마틴, 스테비아 추출물 (예를 들어 레바우디오시드 A, 글리시르히진 등]) 및 합성 향미제(사카린, 아스파르탐 등)를 사용할 수 있다.When the composition of the present invention is made of a food composition, it includes not only the active ingredient of the present invention as described above, but also components commonly added in the preparation of food, and include, for example, proteins, carbohydrates, fats, nutrients, seasonings, and the like. Contains flavors. Examples of the above carbohydrates include monosaccharides such as glucose, fructose and the like; Disaccharides such as maltose, sucrose, oligosaccharides and the like; And sugars such as conventional sugars such as polysaccharides such as dextrin, cyclodextrin and the like and xylitol, sorbitol, erythritol. As the flavoring agent, natural flavoring agents (tauumatin, stevia extract (for example rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be used.
예컨대, 본 발명의 식품 조성물이 드링크제로 제조되는 경우에는 본 발명의 유효성분 이외에 구연산, 액상과당, 설탕, 포도당, 초산, 사과산, 과즙, 두충 추출액, 대추 추출액, 감초 추출액 등을 추가로 포함시킬 수 있다.For example, when the food composition of the present invention is prepared with a drink, in addition to the active ingredient of the present invention, citric acid, liquid fructose, sugar, glucose, acetic acid, malic acid, fruit juice, tofu extract, jujube extract, licorice extract, etc. may be further included. have.
본 발명의 바람직한 구현예에 따르면, 본 발명의 조성물은 화장료 조성물이다.According to a preferred embodiment of the present invention, the composition of the present invention is a cosmetic composition.
본 발명의 화장료 조성물에 포함되는 성분은 상술한 본 발명의 유효 성분 이외에 화장품 조성물에 통상적으로 이용되는 성분들을 포함하며, 예컨대 항산화제, 안정화제, 용해화제, 비타민, 안료 및 향료와 같은 통상적인 보조제, 그리고 담체를 포함한다. 또한, 상기 화장료 조성물은 그 효과를 증진시키기 위하여 피부 흡수 촉진 물질을 추가로 포함할 수 있다.The components included in the cosmetic composition of the present invention include components conventionally used in cosmetic compositions in addition to the active ingredients of the present invention described above, and include conventional auxiliaries such as antioxidants, stabilizers, solubilizers, vitamins, pigments and flavorings. And carriers. In addition, the cosmetic composition may further include a skin absorption promoting substance to enhance the effect.
본 발명의 화장료 조성물은 당업계에서 통상적으로 제조되는 어떠한 제형으로도 제조될 수 있으며, 예를 들어, 용액, 현탁액, 유탁액, 페이스트, 겔, 크림, 로션, 파우더, 비누, 계면활성제-함유 클린싱, 오일, 분말 파운데이션, 유탁액 파운데이션, 왁스 파운데이션 및 스프레이 등으로 제형화될 수 있으나, 이에 한정되는 것은 아니다. 보다 상세하게는, 유연 화장수, 영양 화장수, 영양 크림, 마사지 크림, 에센스, 아이 크림, 클렌징 크림, 클렌징 포옴, 클렌징 워터, 팩, 스프레이 또는 파우더의 제형으로 제조될 수 있다.The cosmetic composition of the present invention may be prepared in any formulation commonly prepared in the art, for example, solutions, suspensions, emulsions, pastes, gels, creams, lotions, powders, soaps, surfactant-containing cleansing , Oils, powder foundations, emulsion foundations, wax foundations and sprays, and the like, but are not limited thereto. More specifically, it may be prepared in the form of a flexible lotion, nutrition lotion, nutrition cream, massage cream, essence, eye cream, cleansing cream, cleansing foam, cleansing water, pack, spray or powder.
본 발명의 제형이 페이스트, 크림 또는 겔인 경우에는 담체 성분으로서 동물성유, 식물성유, 왁스, 파라핀, 전분, 트라칸트, 셀룰로오스 유도체, 폴리에틸렌 글리콜, 실리콘, 벤토나이트, 실리카, 탈크 또는 산화아연 등이 이용될 수 있다.When the formulation of the present invention is a paste, cream or gel, animal oils, vegetable oils, waxes, paraffins, starches, trachants, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicas, talc or zinc oxide may be used as carrier components. Can be.
본 발명의 제형이 파우더 또는 스프레이인 경우에는 담체 성분으로서 락토스, 탈크, 실리카, 알루미늄 히드록시드, 칼슘 실리케이트 또는 폴리아미드 파우더가 이용될 수 있고, 특히 스프레이인 경우에는 추가적으로 클로로플루오로히드로카본, 프로판/부탄 또는 디메틸 에테르와 같은 추진체를 포함할 수 있다.When the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used, in particular in the case of a spray, additionally chlorofluorohydrocarbon, propane Propellant such as butane or dimethyl ether.
본 발명의 제형이 용액 또는 유탁액인 경우에는 담체 성분으로서 용매, 용해화제 또는 유탁화제가 이용되고, 예컨대 물, 에탄올, 이소프로판올, 에틸 카보네이트, 에틸 아세테이트, 벤질 알코올, 벤질 벤조에이트, 프로필렌 글리콜, 1,3-부틸 글리콜 오일, 글리세롤 지방족 에스테르, 폴리에틸렌 글리콜 또는 소르비탄의 지방산 에스테르가 있다.When the formulation of the present invention is a solution or emulsion, a solvent, solubilizer or emulsifier is used as the carrier component, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1 Fatty acid esters of, 3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol or sorbitan.
본 발명의 제형이 현탁액인 경우에는 담체 성분으로서 물, 에탄올 또는 프로필렌 글리콜과 같은 액상의 희석제, 에톡실화 이소스테아릴 알코올, 폴리옥시에틸렌 소르비톨 에스테르 및 폴리옥시에틸렌 소르비탄 에스테르와 같은 현탁제, 미소결정성 셀룰로오스, 알루미늄 메타히드록시드, 벤토나이트, 아가 또는 트라칸트 등이 이용될 수 있다.When the formulation of the present invention is a suspension, liquid carrier diluents such as water, ethanol or propylene glycol, suspending agents such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, microcrystals Soluble cellulose, aluminum metahydroxy, bentonite, agar or tracant and the like can be used.
본 발명의 제형이 계면-활성제 함유 클린징인 경우에는 담체 성분으로서 지방족 알코올 설페이트, 지방족 알코올 에테르 설페이트, 설포숙신산 모노에스테르, 이세티오네이트, 이미다졸리늄 유도체, 메틸타우레이트, 사르코시네이트, 지방산 아미드 에테르 설페이트, 알킬아미도베타인, 지방족 알코올, 지방산 글리세리드, 지방산 디에탄올아미드, 식물성 유, 라놀린 유도체 또는 에톡실화 글리세롤 지방산 에스테르 등이 이용될 수 있다.When the formulation of the present invention is a surfactant-containing cleansing, the carrier component is an aliphatic alcohol sulfate, an aliphatic alcohol ether sulfate, a sulfosuccinic acid monoester, an isethionate, an imidazolinium derivative, a methyltaurate, a sarcosinate, a fatty acid amide. Ether sulfates, alkylamidobetaines, aliphatic alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, lanolin derivatives or ethoxylated glycerol fatty acid esters and the like can be used.
본 발명의 특징 및 이점을 요약하면 다음과 같다:The features and advantages of the present invention are summarized as follows:
(ⅰ) 본 발명의 조성물은 홍삼 추출물, 자소 추출물, 알로에 추출물, 어성초 추출물 및 국화 추출물을 유효성분으로 포함한다.(Iii) The composition of the present invention comprises red ginseng extract, lyso extract, aloe extract, Echochocho extract and chrysanthemum extract as active ingredients.
(ⅱ) 본 발명의 유효성분은 우수한 히스타민 유리 억제 및 IgE 생성 억제 효과를 나타내어 IgE 매개 알레르기 질환의 예방 또는 치료 효과를 가지고, 우수한 림프구 증식, 및 IL-2와 IFN-γ의 증가 효과를 나타내어 T세포 매개 알레르기 질환 의 예방 또는 치료 효과를 가진다.(Ii) The active ingredient of the present invention exhibits an excellent histamine free inhibitory effect and an IgE production inhibitory effect, and thus has an effect of preventing or treating IgE mediated allergic disease, an excellent lymphocyte proliferation, and an increase effect of IL-2 and IFN-γ. It has a prophylactic or therapeutic effect on cell mediated allergic diseases.
(ⅲ) 또한, 홍삼 추출물, 자소 추출물, 알로에 추출물, 어성초 추출물 또는 국화 추출물을 각각 복용하였을 때 보다, 본 발명의 복합 유효성분을 포함하는 조성물을 복용하였을 때 상승적(synergic) 효과를 가진다.(Iii) In addition, it has a synergic effect when the composition containing the composite active ingredient of the present invention is taken, than when taking red ginseng extract, shiso extract, aloe extract, echochocho extract or chrysanthemum extract, respectively.
(ⅳ) 또한, 본 발명의 조성물은 천연물질을 유효성분으로 하여 세포독성 및 피부 부작용이 없어 식품, 약제학적 및 화장료 조성물에 안전하게 적용할 수 있다.(Iii) In addition, the composition of the present invention can be safely applied to food, pharmaceutical and cosmetic compositions as a natural substance as an active ingredient, there is no cytotoxicity and skin side effects.
이상에서 상세히 설명한 바와 같이, 본 발명은 홍삼 추출물, 자소 추출물, 알로에 추출물, 어성초 추출물 및 국화 추출물을 유효성분으로 포함하는 알레르기(allergy) 질환 예방 또는 치료용 조성물을 제공한다. 본 발명의 조성물에서 이용되는 유효성분은 우수한 히스타민 유리 억제, IgE 생성 억제, 림프구 증식, 및 IL-2 및 IFN-γ의 증가 효과를 나타내어 우수한 알레르기 질환의 예방 또는 치료 효과를 가진다. 또한, 홍삼 추출물, 자소 추출물, 알로에 추출물, 어성초 추출물 또는 국화 추출물을 각각 복용하였을 때 보다, 본 발명의 복합 유효성분을 포함하는 조성물을 복용하였을 때 상승적(synergic) 효과를 가진다. 또한, 본 발명의 조성물은 천연물질을 유효성분으로 하여 세포독성 및 피부 부작용이 없어 식품, 약제학적 및 화장료 조성물에 안전하게 적용할 수 있다.As described in detail above, the present invention provides a composition for the prevention or treatment of allergy disease (allergy) comprising red ginseng extract, shiso extract, aloe extract, Echochocho extract and chrysanthemum extract as an active ingredient. The active ingredient used in the composition of the present invention exhibits excellent histamine free inhibition, IgE production inhibition, lymphocyte proliferation, and an increase effect of IL-2 and IFN-γ, thus having an excellent preventive or therapeutic effect of allergic diseases. In addition, it has a synergic effect when taking a composition comprising the complex active ingredient of the present invention, than when taking red ginseng extract, shiso extract, aloe extract, Echochocho extract or chrysanthemum extract, respectively. In addition, the composition of the present invention can be safely applied to foods, pharmaceuticals and cosmetic compositions having a natural substance as an active ingredient, there is no cytotoxicity and skin side effects.
이하, 실시예를 통하여 본 발명을 더욱 상세히 설명 하고자 한다. 이들 실시예는 오로지 본 발명을 보다 구체적으로 설명하기 위한 것으로서, 본 발명의 요지 따라 본 발명의 범위가 이들 실시예에 의해 제한되지 않는다는 것은 본 발명이 속하는 기술 분야에서 통상의 지식을 가진 자에게 있어서 자명할 것이다. Hereinafter, the present invention will be described in more detail with reference to Examples. These examples are only for illustrating the present invention in more detail, and the scope of the present invention is not limited by these examples in accordance with the gist of the present invention to those skilled in the art. Will be self explanatory.
실시예Example
실시예Example 1: 생약재의 1: of herbal medicine 열수Hydrothermal 추출물 제조 Extract manufacturer
항알레르기 활성을 가지는 생약재 추출물을 선별하기 위해 홍삼, 자소, 알로에, 어성초 및 국화를 추출온도 85℃에서 정제수를 이용하여 2회 추출하였으며, 추출배수는 1차 10배, 2차 및 3차는 5배로 추출하여 농축 후 동결 건조하여 분말 시료로 만들어 이용하였다. To select herbal extracts with anti-allergic activity, red ginseng, jaso, aloe, eosungcho and chrysanthemum were extracted twice using purified water at extraction temperature of 85 ℃. Extraction, concentration and freeze drying were used to make a powder sample.
실시예Example 2: 2: 생약재Herbal medicine 추출물의 투여 Administration of Extract
Balb/C 마우스(대한실험동물)에 투여하는 생약재의 양은 사람의 1일 복용량을 기준으로 홍삼 추출물 100 mg/kg, 자소 추출물 2.5 mg/kg, 알로에 추출물 0.5 mg/kg, 어성초 추출물 57.5 mg/kg 또는 국화 추출물 32.5 mg/kg을 1일 식수 섭취량에 섞어 음용시켰다. 실험을 시작하기 2주 전부터 실험이 끝날 때까지 각각의 생약재 추출물을 음용시켰다.The amount of herbal medicine administered to Balb / C mice (Korean experimental animal) was 100 mg / kg of red ginseng extract, 2.5 mg / kg of bovine extract, 0.5 mg / kg of aloe extract, and 57.5 mg / kg of Echo extract. Or 32.5 mg / kg chrysanthemum extract was mixed with daily drinking water intake. Each herbal extract was drunk two weeks before the experiment and until the end of the experiment.
실험예Experimental Example 1: 항알레르기 활성을 가지는 1: have antiallergic activity 생약재Herbal medicine 추출물의 선별 Screening of Extracts
상기 실시예 1에서 제조한 홍삼, 자소, 알로에, 어성초 및 국화 추출물의 항알레르기 활성을 조사하였다.The anti-allergic activity of the red ginseng, Jaso, aloe, eosungcho and chrysanthemum extract prepared in Example 1 was investigated.
이를 위해 Balb/C 마우스를 대상으로 2,4-디니트로클로로벤젠(DNCB, Sigma)를 이용하여 실험적인 면역과민반응을 유발하였다. 실험에 이용할 마우스는 대조군 및 DNCB로 면역과민반응을 유발한 군으로 구분하고, 면역과민반응은 이개 부종 시험(ear swelling test)으로 1차 검증하여 항알레르기 활성을 가지는 생약재 추출물을 선별하였다.For this purpose, experimental immune hypersensitivity reactions were induced using 2,4-dinitrochlorobenzene (DNCB, Sigma) in Balb / C mice. The mice to be used in the experiments were divided into control and DNCB-induced immune hypersensitivity reactions. The immune hypersensitivity reactions were first verified by an ear swelling test to select herbal extracts having anti-allergic activity.
1-1) DNCB 면역법 1-1) DNCB Immunology
DNCB(Simga)를 이용 직전에 만들어 합텐으로 이용하였다. DNCB는 아세톤 : 올리브 오일 (4:1) 용액에 원하는 농도로 용해시켜 이용하였다. DNCB 피부 감작을 위해 Balb/C 마우스의 복부 털을 조심스럽게 제거하여 노출된 경부, 복부, 액와 및 서혜부의 피부에 1% DNCB를 각각 50 ㎕씩 도포하였다.DNCB (Simga) was made just before use and used as hapten. DNCB was dissolved in acetone: olive oil (4: 1) solution at the desired concentration. The abdominal hair of Balb / C mice was carefully removed for DNCB skin sensitization, and 50 μl of 1% DNCB was applied to the exposed cervical, abdominal, axillary and inguinal skin, respectively.
1-2) 이개 부종(Ear swelling) 측정법1-2) Ear swelling measurement
면역과민반응의 유발은 DNCB 감작 5일 후에 25% DNCB 50 ㎕를 귀에 도포하고 24시간 경과한 후 오른쪽 귀의 두께를 버티칼 캘리퍼(Mitutoyo, Japan)로 측정한 다음 이개 부종의 평균값을 구하여 각각의 차이를 비교하였다. 대조군으로는 아세톤 : 올리브 오일 용액만을 동일한 방식으로 처리하여 비교하였다.To induce the immune hypersensitivity reaction, 50 μl of 25% DNCB was applied to the ear 5 days after DNCB sensitization, and after 24 hours, the thickness of the right ear was measured with a vertical caliper (Mitutoyo, Japan), and then the average value of the edema was determined. Compared. As a control, only acetone: olive oil solution was treated in the same manner and compared.
실시예Example 3: 복합 생약 조성물의 제조 3: Preparation of complex herbal composition
상기 실험예 1에서 선별된 홍삼, 자소, 알로에, 어성초 및 국화 열수추출물을 하기 표 1의 조성에 따라 혼합하여, 복합추출물을 제조하였다.Red ginseng, jaso, aloe, eoseongcho and chrysanthemum hydrothermal extracts selected in Experimental Example 1 were mixed according to the composition of Table 1 below to prepare a composite extract.
실험예Experimental Example 2: 2: TypeType I 알레르기에 대한 효과 조사 I Examination of effects on allergies
2-1) 히스타민 유리 억제 효과 조사2-1) Histamine release inhibitory effect investigation
본 발명의 조성물에 의한 히스타민 유리 억제효과를 실험하였다. 우선, 하기 표 3의 생약재 추출물 각각을 Balb/C 마우스에 실험을 시작하기 2주 전부터 실험이 끝날 때까지 음용시켰다. 실시예 3에서 제조된 복합 생약 조성물의 투여량은 각각, 복합물(X 0.5) 50 mg/kg, 복합물(X 1.0) 100 mg/kg 또는 복합물(X 5.0) 500 mg/kg이다.The histamine release effect by the composition of the present invention was tested. First, each of the herbal extracts of Table 3 was drunk in Balb / C mice two weeks before the experiment was started until the end of the experiment. The dosage of the composite herbal composition prepared in Example 3 is 50 mg / kg of complex (X 0.5), 100 mg / kg of complex (X 1.0) or 500 mg / kg of complex (X 5.0), respectively.
이어, OVA(obalbumin, Sigma)로 3차 면역 후 2일 뒤에 마우스의 전혈(whole blood) 0.5 ml를 채취하고, 여기에 0.5 N HClO4 2 ml를 첨가하여 진탕 후 원심분리(12,000 rpm, 20 min 및 4℃)하였다. 상층액 2 ml를 분리하여 증류수 3 ml 및 n-부탄올 10 ml를 첨가하고, 고체 NaCl를 침전이 생겨 더 이상 녹지 않을 때까지 볼텍싱을 해주면서 조금씩 첨가한 후 6 N NaOH을 0.2 ml 첨가하여 원심분리 하였다. 원심 분리 결과 얻어진 부탄올 층 3 ml에 n-헵탄 3 ml와 0.1 N HCl 1.2 ml를 넣고 100℃ 워터배스에서 10분 동안 방치한 후 원심분리(3,000 rpm, 15 min 및 4℃)하였다. 또한, 원심 분리 결과 얻어진 HCl층 1 ml에 1 N NaOH 0.15 ml와 0.2% OPT(o-phthaldialdehyde, Sigma) 0.1 ml를 넣고 0℃에서 40분 동안 반응시킨 후 0.5 N H2SO4 0.14 ml를 첨가하고 히스타민(0-200 mg)/0.1 N HCl 1 ml를 분주한 후 1 N NaOH 0.15 ml와 0.2% OPT 0.1 ml를 첨가한 후 0℃에서 40분 동안 반응시켰다. 이에 0.5 N H2SO4 0.14 ml를 첨가하고 형광강도측정기(FP-6200, JASCO, 일본)를 이용하여 여기(excitation) 파장 350 nm와 방출(emission) 파장 440 nm에서 석영 큐벳을 사용하여 형광광도를 측정하였다. 히스타민 분비의 억제율은 다음의 수학식 1을 이용하여 계산하였다:Then, 2 days after the third immunization with OVA (obalbumin, Sigma), 0.5 ml of whole blood of the mouse was collected, and 2 ml of 0.5 N HClO 4 was added thereto, followed by centrifugation after shaking (12,000 rpm, 20 min). And 4 ° C.). 2 ml of the supernatant was separated and 3 ml of distilled water and 10 ml of n-butanol were added, and the solid NaCl was added little by little while vortexing until a precipitate formed and dissolved, and then 0.2 ml of 6 N NaOH was added and centrifuged. It was. 3 ml of n-heptane and 1.2 ml of 0.1 N HCl were added to 3 ml of the butanol layer obtained by centrifugation, and the mixture was left for 10 minutes in a 100 ° C. water bath, followed by centrifugation (3,000 rpm, 15 min and 4 ° C.). In addition, 0.15 ml of 1 N NaOH and 0.1 ml of 0.2% OPT (o-phthaldialdehyde, Sigma) were added to 1 ml of the HCl layer obtained as a result of centrifugation, and reacted at 0 ° C. for 40 minutes, and then 0.14 ml of 0.5 NH 2 SO 4 was added thereto. After dispensing 1 ml of histamine (0-200 mg) /0.1 N HCl, 0.15 ml of 1 N NaOH and 0.1 ml of 0.2% OPT were added and reacted at 0 ° C. for 40 minutes. 0.14 ml of 0.5 NH 2 SO 4 was added thereto, and the fluorescence intensity was measured using a quartz cuvette at an excitation wavelength of 350 nm and an emission wavelength of 440 nm using a fluorescence intensity meter (FP-6200, JASCO, Japan). Measured. The inhibition rate of histamine secretion was calculated using Equation 1 below:
상기 표 2에서 볼 수 있듯이, 본 발명의 유효성분인 복합물을 처리한 후 OVA를 처리한 경우의 히스타민 유리 억제율은 72%로 매우 높게 나타났다. 이는 각각의 생약재를 단독으로 처리한 경우에 비해 매우 높으며, 현재 알레르기 치료제로 사용 중인 지르텍과 비슷한 결과를 보임을 알 수 있다. 따라서 본 발명의 조성물은 히스타민에 의해서 유발되는 피부 알레르기의 예방 또는 치료에 우수한 효과를 보임을 확인 할 수 있다.As can be seen in Table 2, the histamine release rate was very high as 72% when the OVA was treated after the composite of the present invention. This is very high compared with the treatment of each herbal alone, it can be seen that the results similar to Zirtec currently used as an allergy treatment. Therefore, the composition of the present invention can be confirmed to show an excellent effect in the prevention or treatment of skin allergy caused by histamine.
2-2) 본 발명 유효성분인 복합물의 IgE 생성 억제 효과 조사2-2) Investigation of the inhibitory effect of IgE production of the composite of the present invention
IgE는 조직 내 염증반응과 깊은 관련이 있으며, 마스트 세포(mast cell)의 Fc 수용체와 가교결합을 통해 히스타민과 에코사노이드(eicosanoid) 등을 생성시켜 알레르기 증상을 유발시키는 항체로서 잘 알려져 있다. 따라서 과민성 질환 치료 효과가 있는지 여부를 알아보기 위해 상기 본 발명의 생약 복합물이 과민성 질환에 있어서 대표적인 과민성 유발 요인인 IgE의 생성량에 미치는 영향을 실험하였다.IgE is closely related to inflammatory reactions in tissues and is well known as an antibody causing allergic symptoms by producing histamine and ecosanoids through crosslinking with Fc receptors in mast cells. Therefore, the effects of the herbal complex of the present invention on the production of IgE, which is a representative sensitization inducing factor in sensitizing diseases, were examined to determine whether there is a treatment effect for sensitizing diseases.
혈청의 IgE 함량은 ELISA 법에 의해 시행하였다. 코팅 완충액에 항 마우스 IgE(BD PharMingen)를 2 ㎍/ml의 농도로 희석해서 웰 당 100 ㎕씩 분주 하고 4℃에서 하룻밤 동안 코팅한 다음, 혈청을 10% FBS/PBS(또는 3% BSA/PBS)에 적절한 농도로 희석해서 100 ㎕씩 분주하고 상온에서 1시간 반응시켰다. 그 후 플레이트를 PBS/Tween으로 3회 세척하고 2 ㎍/ml의 농도로 희석한 바이오틴이 결합된 2차 항체(anti-mouse-IgE)(BD PharMingen)를 100 ㎕씩 넣고 상온에서 1시간 반응시키고, 이를 다시 세척한 후 아비딘-페록시다아제(BD PharMingen)를 분주하고 실온에서 30분 반응시켰다. 플레이트를 6회 세척한 후 ABTS(2,2'-azino-bis-(3-ethylbenz thiazoline-6-sulfonic acid), BD PharMingen) 기질 용액에 H2O2를 1000배 되게 희석해서 웰에 100 ㎕씩 분주하고 ELISA 리더(Skatron, 노르웨이) 로 405 nm에서 흡광도를 측정하였다. 하기 수학식 2를 이용하여 IgE 생성 억제율을 계산하였다:Serum IgE content was performed by ELISA method. Dilute anti-mouse IgE (BD PharMingen) to a concentration of 2 μg / ml in coating buffer and dispense 100 μl per well and coat overnight at 4 ° C., then serum 10% FBS / PBS (or 3% BSA / PBS) ) Was diluted to an appropriate concentration, and 100 µl was dispensed and reacted at room temperature for 1 hour. Then, wash the plate three times with PBS / Tween, add 100 μl of biotin-bound secondary antibody (anti-mouse-IgE) (BD PharMingen) diluted to a concentration of 2 μg / ml, and react for 1 hour at room temperature. After washing again, avidin-peroxidase (BD PharMingen) was dispensed and reacted at room temperature for 30 minutes. After washing the plate six times, 100 μl of the wells were diluted 1000 times with H 2 O 2 in ABTS (2,2'-azino-bis- (3-ethylbenz thiazoline-6-sulfonic acid), BD PharMingen) substrate solution. Aliquots were taken and absorbance was measured at 405 nm with an ELISA reader (Skatron, Norway). Inhibition of IgE production was calculated using Equation 2 below:
상기 표 3에서 확인 할 수 있듯이, 본 발명의 유효성분인 생약 복합물은 과민성 대표 인자인 IgE의 생성을 52% 억제시킴을 확인하였으며, 현재 알레르기 치료제로 사용 중인 지르텍에 비해서도 IgE의 생성을 억제시키는 효과가 뛰어남을 확인하였다. 상기 실험결과를 통하여, 본 발명의 유효성분인 생약 복합물은 IgE 매개 알레르기 질환에 대한 우수한 예방 또는 치료 효과를 가짐을 확인 할 수 있다.As can be seen in Table 3, it was confirmed that the herbal compound as an active ingredient of the present invention inhibits the generation of hypersensitivity representative factor IgE by 52%, the effect of inhibiting the production of IgE compared to Zirtec currently used as an allergy treatment Confirmed excellent. Through the above experimental results, it can be confirmed that the herbal compound as an active ingredient of the present invention has an excellent prophylactic or therapeutic effect against IgE mediated allergic diseases.
실험예Experimental Example 3: 3: TypeType IVIV 알레르기에 대한 효과 조사 Investigate the effects on allergies
3-1) 이개부종 억제반응 조사3-1) Investigation of inhibition of edema
면역과민반응의 유발은 DNCB 감작 5일 후에 0.25% DNCB 50 ㎕를 귀에 도포하고 24시간 경과 후 오른쪽 귀의 두께를 버티칼 캘리퍼(Mitutoyo, 일본)로 측정하여 귀 두께 평균값을 구하여 각각의 차이를 비교하였다. 대조군으로 아세톤 : 올리브 오일 용액만을 동일한 방식으로 처리하여 비교하였다. 그 결과를 하기의 표 4에 나타내었다. Induction of immune hypersensitivity reaction was applied to 50 μl of 0.25% DNCB after 5 days of DNCB sensitization, and after 24 hours, the thickness of the right ear was measured with a vertical caliper (Mitutoyo, Japan) to obtain the average thickness of the ears and compared the differences. As a control, only acetone: olive oil solution was treated in the same manner and compared. The results are shown in Table 4 below.
상기 표 4에서 확인 할 수 있듯이, 본 발명의 유효성분인 생약 복합물이 이개부종반응을 가장 많이 억제함을 알 수 있었으며, 현재 알레르기 치료제로 사용 중인 지르텍에서는 억제효과를 기대하기 어려운 결과를 얻었다.As can be seen in Table 4, it was found that the herbal compound as an active ingredient of the present invention most inhibits the edema reaction, Zirtec is currently used as an allergy treatment agent, it was difficult to expect the inhibitory effect.
3-2) 림프구 증식 및 사이토카인(IL-2, IFN-γ)에 미치는 영향 조사3-2) Investigation of the effects on lymphocyte proliferation and cytokines (IL-2, IFN-γ)
림프구 증식에 미치는 영향을 조사하기 위하여, 마우스의 림프절을 무균적으로 적출하여 기계적으로 파쇄한 다름, 거즈로 여과하여 단일 세포부유액을 만들었다. 이 림프절 세포부유액을 3회 세척한 다음, 25 mM HEPES, 100 units/㎖ 페니실린, 100 ㎍/㎖ 스트렙토마이신 및 10% 열 불활성화 우 태아 혈청이 첨가된 RPMI-1640 배양액 (Gibco, Renfrewshire, 영국)으로 적당한 농도가 되게 희석하였다. 살아있는 세포의 계수는 트리판 블루 다이 익스클루션 법을 이용하였다. 림프구 증식반응을 측정하기 위하여 96 웰 평판 배양 플레이트에 세포현탁액을 2.5× 105 cells/well 이 되도록 첨가하고, 자극제로 콘카나발린-A(ConA)(Sigma)를 첨가하여 37℃ 및 5% CO2 배양기에서 5일간 배양하였다. 배양 종료 4시간 전에 셀 티터 용액(Promega)을 웰 당 25 ㎕씩 넣어 주었다. 배양종료 후 마이크로플레이트 리더(Skatron, 노르웨이)로 490 nm에서 흡광도를 측정하였다. 결과는 평균± 표준편차(SD)로 표시하였다.To investigate the effect on lymphocyte proliferation, lymph nodes in mice were aseptically extracted and mechanically disrupted, filtered with gauze to produce single cell suspensions. The lymph node cell suspension was washed three times, followed by RPMI-1640 culture (Gibco, Renfrewshire, UK) with 25 mM HEPES, 100 units / ml penicillin, 100 μg / ml streptomycin and 10% heat inactivated fetal bovine serum. Diluted to an appropriate concentration. Counting of living cells was carried out using trypan blue die extrusion method. To measure lymphocyte proliferation, add cell suspension to 2.5 × 10 5 cells / well in a 96 well plate culture plate and add Concanavalin-A (ConA) (Sigma) as a stimulant to 37 ° C. and 5% CO. Incubated for 5 days in two incubators. 4 μl of cell titer solution (Promega) was added to each well 4 hours before the end of the culture. After incubation, the absorbance at 490 nm was measured with a microplate reader (Skatron, Norway). The results are expressed as mean ± standard deviation (SD).
사이토카인의 농도 측정은 ELISA법을 이용하여 결정하였다. 즉, 평판 마이크로웰 플레이트에 염소 항 마우스(goat anti-mouse) IL-2 및 IFN-γ 항체(BD PharMingen)를 코팅 완충액(PBS)을 이용하여 4℃에서 하룻밤 반응 시킨 다음, 3% BSA 용액으로 30분 동안 상온에서 블록킹 하였다. 실험에서 채취한 혈청을 적당한 비율로 희석하여 플레이트에 각각 넣고, 실온에서 1시간 반응 시킨 다음 바이오틴 표지 항 IL-2 및 IFN-γ를 첨가하였다. 그리고 아비딘-컨쥬게이트 알카라인 포스페이트(Zymed)를 적량 가하고 실온에서 1시간 반응시키고, 기질로 p-니트로페닐 포스페이트(BD PharMingen)를 넣은 다음 마이크로플레이트 리더(Skatron, 노르웨이)로 410 nm와 450 nm에서 흡광도를 측정하여 표준 곡선을 이용하여 ng/㎖ 농도로 환산하여 나타내었다. 림프구 증식, IL-2 및 IFN-γ에 대한 측정결과를 표 5에서 비교하였다. Measurement of the concentration of cytokines was determined using the ELISA method. In other words, goat anti-mouse (goat anti-mouse) IL-2 and IFN-γ antibody (BD PharMingen) was reacted overnight at 4 ° C using a coating buffer (PBS) on a plate microwell plate, followed by 3% BSA solution. Blocking at room temperature for 30 minutes. Serum collected in the experiment was diluted to an appropriate ratio and put into plates, respectively, and reacted at room temperature for 1 hour, and then biotin-labeled anti IL-2 and IFN-γ were added. Avidin-conjugated alkaline phosphate (Zymed) was added in an appropriate amount and reacted at room temperature for 1 hour, and p-nitrophenyl phosphate (BD PharMingen) was added as a substrate, followed by absorbance at 410 nm and 450 nm with a microplate reader (Skatron, Norway). Was measured and expressed in terms of ng / ml using a standard curve. The results of lymphocyte proliferation, IL-2 and IFN-γ were compared in Table 5.
상기 표 5에서 볼 수 있듯이, 림프구 증식, IL-2 및 IFN-γ는 본 발명의 유효성분인 복합물 처리군에서 단일추출물 처리군보다 높게 나타남을 확인 할 수 있다. 하여, 본 발명의 유효성분인 생약 복합물은 T세포 매개 알레르기 질환을 예방 또는 치료하는데 우수한 효과를 가짐을 알 수 있다.As can be seen in Table 5, it can be confirmed that lymphocyte proliferation, IL-2 and IFN-γ are higher than the single extract treatment group in the complex treatment group of the active ingredient of the present invention. Thus, it can be seen that the herbal compound as an active ingredient of the present invention has an excellent effect in preventing or treating T cell mediated allergic diseases.
실시예Example 4: 음료의 제조 4: manufacture of beverage
실시예 3에서 제조된 생약추출물 20%, 비타민 B1 0.01%, 비타민B2 0.01%, 비타민 C 0.45%, L-루이신 0.2%, L-이소루이신 0.15%, L-발린 0.15%, 타우린 0.15%, 당류 11.5% 및 정제수 잔여량을 혼합하여 음료 조성물을 제조하였다.20% herbal extract prepared in Example 3, vitamin B1 0.01%, vitamin B2 0.01%, vitamin C 0.45%, L- leucine 0.2%, L- isoleucine 0.15%, L- valine 0.15%, taurine 0.15% , 11.5% of saccharides and the residual amount of purified water were mixed to prepare a beverage composition.
실시예Example 5: 과립의 제조 5: Preparation of Granules
실시예 3에서 제조한 생약추출물 분말 35 중량%, 비타민C 0.2 중량%, 포도당 28 중량%, 덱스트린 28 중량%, 70% 에탄올 8.7중량%를 함께 반죽하고 과립제조기를 통과시켜 과립 60 mesh의 과립을 제조하였다.35% by weight of the herbal extract powder prepared in Example 3, 0.2% by weight of vitamin C, 28% by weight of glucose, 28% by weight of dextrin, 8.7% by weight of 70% ethanol were kneaded together and passed through a granulator to obtain granules of granule 60 mesh. Prepared.
실시예Example 6: 정제의 제조 6: manufacture of tablets
실시예 3에서 제조한 분무건조 분말 35 중량%, 유당 49.5 중량%, 결정셀룰로오즈 8 중량%, 실리콘 알루민산나트륨 3 중량%, 자당 지방산 에스테르 2 중량%, 코팅제 및 식용색소 2.5 중량%를 혼합하여 타정기로 정제를 제조하였다. 35 wt% of the spray dried powder prepared in Example 3, 49.5 wt% of lactose, 8 wt% of crystalline cellulose, 3 wt% of sodium aluminate, 2 wt% of sucrose fatty acid ester, coating agent and 2.5 wt% of food coloring. Tablets were prepared.
실시예Example 7: 연질캡슐의 제조 7: Preparation of soft capsule
실시예 3에서 제조한 분무건조 분말 30 중량%, 대두유 63 중량%, 백납 4.05 중량%, α-토코페롤 2.945 중량% 및 비타민 D3 0.005 중량%를 혼합하고 균질화하여 유화시킨 후 젤라틴으로 피막을 형성시키고 항온항습실에서 14일간 건조시킨 후 선별, 포장하여 연질캡슐을 제조하였다.30% by weight of the spray dried powder prepared in Example 3, 63% by weight of soybean oil, 4.05% by weight of lead, 2.945% by weight of α-tocopherol, and 0.005% by weight of vitamin D3 were mixed, homogenized and emulsified to form a film with gelatin and constant temperature. After drying for 14 days in a humidity chamber, soft capsules were prepared by screening and packaging.
이상으로 본 발명의 특정한 부분을 상세히 기술하였는 바, 당업계의 통상의 지식을 가진 자에게 있어서 이러한 구체적인 기술은 단지 바람직한 구현예일 뿐이며, 이에 본 발명의 범위가 제한되는 것이 아닌 점은 명백하다. 따라서, 본 발명의 실질적인 범위는 첨부된 청구항과 그의 등가물에 의하여 정의된다고 할 것이다.Having described the specific part of the present invention in detail, it is apparent to those skilled in the art that the specific technology is merely a preferred embodiment, and the scope of the present invention is not limited thereto. Thus, the substantial scope of the present invention will be defined by the appended claims and equivalents thereof.
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| CN109394987A (en) * | 2018-12-06 | 2019-03-01 | 中山大学中山眼科中心 | The composition and preparation method thereof with treatment eye disease containing dimethyl fumarate |
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