KR100881445B1 - Skin protection and skin disease improvement composition and method for producing the same - Google Patents
Skin protection and skin disease improvement composition and method for producing the same Download PDFInfo
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Abstract
본 발명은 유효성분으로서, 당귀, 천궁, 생지황, 하수오, 황기, 감초, 길경, 포공영, 백작약, 백출, 갈근, 박하, 사인, 진피, 율무 및 건율을 혼합한 후 물로 추출하거나 각각 물로 추출한 후 혼합하여 얻은 수추출물 또는 그의 건조분말을 함유하는 피부보호 및 피부질환 개선용 조성물 및 그 제조방법에 관한 것이다.The present invention as an active ingredient, Angelica, Cheonggung, Saenghwang, Sewao, Astragalus, Licorice, Gilgyeong, Pogongyoung, Baekjak, Baekchul, Brown root, Peppermint, Sine, Dermis, Yulmu and dry ratio, and then extracted with water or extracted with water and then mixed The present invention relates to a composition for protecting skin and improving skin diseases containing a water extract or a dry powder thereof, and a method for producing the same.
Description
도 1은 본 발명에 따른 조성물의 인터페론-γ 생성 효과를 보여주는 그래프이다.1 is a graph showing the interferon-γ production effect of the composition according to the present invention.
본 발명은 피부보호 및 피부질환 개선용 생약 조성물에 관한 것으로서, 보다 상세하게는, 유효성분으로서, 당귀, 천궁, 생지황, 하수오, 황기, 감초, 길경, 포공영, 백작약, 백출, 갈근, 박하, 사인, 진피, 율무 및 건율(말린 밤)을 혼합한 후 물로 추출하거나 각각 물로 추출한 후 혼합하여 얻은 수추출물 또는 그의 건조분말을 함유하는, 아토피 피부염 등의 알레르기 피부질환, 및 건선, 습진, 여드름을 포함하는 피부염증 질환은 물론 노화 등에 의한 피부 주름 등을 개선하기 위한 조성물 및 그의 제조방법에 관한 것이다.The present invention relates to a herbal composition for skin protection and skin disease improvement, and more specifically, as an active ingredient, Angelica, Cheongung, Saengjihwang, Shou, Astragalus, Licorice, Giltyeong, Pogongyoung, Baekjak, Baekchul, Brown root, Mint, Sign Allergic skin diseases, such as atopic dermatitis, containing water extracts or dried powders thereof, which are mixed with water, dermis, silk radish and dry rate (dried balm) and then extracted with water or mixed with water, and psoriasis, eczema, acne The present invention relates to a composition for improving skin wrinkles and the like caused by aging and the like, as well as to dermatitis diseases.
인체의 피부는 물리· 화학적으로 외계로부터 신체를 보호하는 동시에 전신의 대사에 필요한 생화학적 기능을 영위하는 생명유지에 필수 불가결한 기관이다. 사람의 머리털, 손· 발톱을 포함한 피부에 나타나는 모든 이상 소견을 피부질환, 즉 피부병이라 한다. 피부는 신체의 표면을 덮고 있으므로 외계로부터 자극이나 여러 병원체에 직접 접촉될 기회가 많고, 체내로부터 큰 영향을 받는다. 더욱이 피부의 근소한 변화도 눈으로 보고 손으로 만질 수 있으며, 병변부의 일부를 채취하여 병리조직학적으로 검사하거나 미생물의 검색 등과 같은 검사를 하기가 쉬워, 개개 질병의 진단을 명확하게 하는 것이 용이하므로, 그 종류는 다른 장기의 것에 비하여 상당한 수에 이르며 병명도 복잡 다양하다. 비교적 흔한 피부질환에는 아토피 피부염, 접촉피부염, 지루성 피부염, 두드러기, 무좀, 완선, 건선, 단순· 대상포진, 피부건조증, 주부습진, 여드름 등이 있다. The skin of the human body is an indispensable organ for the maintenance of life, which physically and chemically protects the body from extraterrestrial and performs the biochemical functions necessary for metabolism of the whole body. All abnormalities that appear on the skin, including human hair, hands and toenails, are called skin diseases, or skin diseases. Since the skin covers the surface of the body, there are many opportunities for direct contact with stimuli or various pathogens from the outside, and are greatly affected by the body. In addition, even minor changes of the skin can be seen by eye and touched by hand, and it is easy to perform pathological histological examination and examination of microorganisms by taking a part of the lesion, and to easily diagnose the diagnosis of individual diseases. The type is considerably larger than that of other organs, and the disease name is complicated. Relatively common skin diseases include atopic dermatitis, contact dermatitis, seborrheic dermatitis, urticaria, athlete's foot, psoriasis, psoriasis, herpes simplex, shingles, dry skin, housewives and acne.
이러한 피부질환의 내인으로는 위장장애· 간장병· 신장병· 물질대사장애· 혈액질환· 내장종양· 내분비장애 등을 들 수 있고, 외인으로는 외계로부터 직접 피부에 작용하는 마찰· 압박· 타박 등의 기계적 자극을 비롯하여, 동물성· 식물성· 광물성의 독물, 그 밖에 약품· 방사선· 온열· 한랭, 또는 세균· 사상균· 원충· 바이러스 등의 미생물이나 피부기생동물 등을 들 수 있다. 또한 유전성 피부질환 및 모반(母斑)· 모반증 등은 그 원인이 유전자에 의한 것이므로 내인성 피부병에 포함된다.Inner skin diseases include gastrointestinal disorders, liver disease, kidney disease, metabolic disorders, blood diseases, visceral tumors, and endocrine disorders.Inners such as friction, pressure, and bruises act on the skin directly from the outside world. In addition to stimulation, animal, vegetable, mineral poisons, other drugs, radiation, heat, cold, microorganisms such as bacteria, filamentous fungi, protozoa, virus, and skin parasites. In addition, hereditary skin diseases and birthmarks and birthmarks are caused by genes and are included in endogenous skin diseases.
피부질환 중, 특히 아토피 피부염은 알레르기 반응으로 인한 질환으로 이러한 알레르기 반응은 초기의 특이적 면역반응과 후기의 염증반응으로 나뉜다. 알레르기 반응은 거의 대부분 비만세포를 매개로 하여 일어나며, 세포막에 존재하는 고친화성의 IgE 수용체(FcεRI)를 통해서 활성화된다. IgE 수용체에 결합되어 있는 IgE 항체가 항원에 의하여 가교(bridge)를 형성하면 포스포리파아제 C, 단백질 인산화효소 C, 칼슘 이온의 작용을 거쳐 과립에 저장되어 있던 히스타민(histamine), 콘드로이틴(chondroitin) 황산염, 헤파린(heparin), 단백질 분해효소 등이 유리됨으로써 반응 초기 단계를 매개한다. 히스타민을 포함한 화학 전달물질들은 알레르기 반응의 초기에 볼 수 있는 여러 가지 임상증상을 유발하는 원인물질들이며, 가장 많은 양을 차지하는 것은 히스타민이다. 따라서 알레르기 반응에서 히스타민에 의한 작용이 중요하며, 그 증상으로는 혈관 확장, 부종 등이 있다. 프로스타글란딘(prostaglandin), 혈소판 활성인자(platelet-activating factor) 등도 알레르기 반응에서 주목받고 있는 화학 전달물질들이다. 화학 전달물질의 작용으로 인하여 나타나는 증상으로는 담마진, 소양증, 피부발적 등의 피부증상을 들 수 있다. Among skin diseases, especially atopic dermatitis is a disease caused by an allergic reaction. The allergic reaction is divided into early specific immune response and late inflammatory response. Most allergic reactions occur through mast cells and are activated through high-affinity IgE receptors (FcεRI) on the cell membrane. When the IgE antibody bound to the IgE receptor forms a bridge by the antigen, histamine and chondroitin sulfate stored in the granules through the action of phospholipase C, protein kinase C and calcium ions Heparin, protease and the like are released to mediate the initial stages of the reaction. Chemical transporters, including histamines, are the causative agents of various clinical symptoms seen early in the allergic reaction, with histamines accounting for the largest amount. Therefore, the action by histamine in the allergic reaction is important, the symptoms include vasodilation, edema. Prostaglandin and platelet-activating factor are also known chemical transporters for allergic reactions. Symptoms resulting from the action of a chemical delivery agent may include skin symptoms such as dimples, pruritis, and skin redness.
현재 환경오염, 식생활 습관 및 유전적인 원인 등으로 피부 트러블 및 피부질환 환자수가 계속적으로 증가하고 있다. 미국의 경우 전 인구의 5∼12%가 피부질환으로 고통 받고 있으며 우리나라의 경우에도 약 200∼300만 명의 피부질환 환자가 있다. 그럼에도 불구하고 현재 그 치료제나 개선제로는 심각한 부작용을 나타내는 스테로이드 제제가 대부분이고, 최근에는 면역억제제가 개발되어 있으나, 아직까지는 부작용 염려 없이 완치 효과를 발휘하는 특별한 치료제나 개선제는 개발되어 있지 않은 실정이다. 이러한 피부질환 치료제의 세계 시장규모는 13억 달러에 달하고 있지만, 면역기능 저하, 피부위축, 어린이 성장방해, 모세혈관 확장, 여드름, 부신피질기능 억제 등 부작용이 수반되므로 임상적 사용에 매우 주의하지 않으면 안 된다. 또한, 현재 항염증 또는 피부질환 치료효과 등이 알려진 생약들 의 유효성분을 추출하여 피부질환 치료제로 개발된 예들이 알려져 있으나, 대부분 가려움증이나 염증 등을 경감시켜 줄 뿐 질병을 완치시키지는 못한다. Currently, the number of patients with skin problems and skin diseases continues to increase due to environmental pollution, dietary habits and genetic causes. In the United States, 5-12% of the population suffers from skin diseases, and in Korea, there are about 2 to 3 million skin disease patients. Nevertheless, most of the steroids that have serious side effects are presently used as therapeutic agents or ameliorators. Recently, immunosuppressive agents have been developed, but there are no special therapeutic agents or ameliorators that have cured effects without concern for side effects. . Although the global market for these skin disease treatments is worth US $ 1.3 billion, it is accompanied by side effects such as decreased immune function, skin atrophy, disruption of children, expansion of capillaries, acne and adrenal cortex, Can not be done. In addition, there are known examples of the development of anti-inflammatory or skin disease, such as the active ingredient extracted from the known herbal medicines for the treatment of skin diseases, but mostly to relieve itching and inflammation, but does not cure the disease.
한편, 생리적 자연현상인 세포 노화로 인해 피부를 구성하고 있는 기본 단위인 피부 세포의 분열이 정상적으로 이루어지지 않고 재생기간이 점차 길어져 피부가 탄력을 잃으며, 노화 각질이 쌓이고 피부가 건조해지면서 주름이 생기기 시작한다. 현재, 노화로 인한 피부 주름을 회복시키기 위해 수술이나 연고 및 비타민 C 요법 등을 사용하고 있지만 인간의 오랜 숙원인 팽팽하고 탄력 있는 피부를 유지시키기에는 역부족이다. 또한, 수 년 전부터 과일 산이나 김치 추출물과 같은 천연물을 이용한 피부 노화 방지 및 피부 개선제가 개발되고 있으나, 남성호르몬인 안드로겐 억제작용 및 그에 따른 피지분비 증가 억제작용에 의한 피부 개선제로서의 기능을 수행할 뿐 피부질환에 대한 치료제로서의 효과는 전혀 나타내지 못하고 있는 실정이다.On the other hand, due to aging of the cells, physiological natural phenomena, skin cells, which are the basic units that make up the skin, are not normally divided, and the regeneration period is gradually longer, and the skin loses its elasticity. It starts to occur. Currently, surgery or ointment and vitamin C therapy are used to repair skin wrinkles due to aging, but it is not enough to maintain a firm and elastic skin, which is a long-time desire of human beings. In addition, skin anti-aging and skin improver using natural products such as fruit acid or kimchi extract have been developed for many years, but it only functions as skin improver by inhibiting androgen, a male hormone, and increasing sebum secretion. There is no effect as a therapeutic agent for skin diseases.
최근에는 의학적 치료와 스킨 케어(skin care)가 접목된, 소위 "메디칼 스킨 케어(medical skin care)"가 우리나라뿐만 아니라 미국, 유럽, 일본 등지에서 피부 치료를 위해 크게 발전해가고 있다. 그러나 겉으로 드러나는 여드름, 기미, 주근깨 등을 개선시킬뿐 알레르기 피부질환이나 면역기능 저하 및 노화 등으로 인한 질환에는 효과가 불분명하다.In recent years, so-called "medical skin care", which combines medical treatment and skin care, has been greatly developed for skin treatment not only in Korea, but also in the United States, Europe and Japan. However, it only improves the appearance of acne, blemishes, freckles, etc. It is unclear for allergic skin diseases, diseases caused by reduced immune function and aging.
한편, 면역세포에서 분비되는 인터페론-γ는 단구와 대식세포에서 항원 특이적· 비특이적 면역반응을 통해 면역조절능을 증가시키는 중요한 역할을 한다. 인터페론-γ 분비가 감소되면 IgE 생성 증가 또는 항원 제거 능력 감소로 아토피 증 상이 심해진다. 또한 인터페론-γ는 T 림프구에 작용하여 이들의 분화와 성숙을 증진시킨다. 또한 CD4 T 림프구에서 Th1 림프구의 분화를 촉진시키며, Th2 림프구의 증식을 억제한다. 이러한 효과는 IL-12를 분비하도록 단핵식균세포를 활성화시키고, T 림프구가 기능적인 IL-12 수용체를 발현하도록 활성화시킴으로써 매개된다. Meanwhile, interferon-γ secreted by immune cells plays an important role in increasing immunomodulatory capacity through antigen-specific and nonspecific immune responses in monocytes and macrophages. Decreased interferon-γ secretion exacerbates atopic symptoms due to increased IgE production or reduced antigenic clearance. Interferon-γ also acts on T lymphocytes to enhance their differentiation and maturation. It also promotes the differentiation of Th1 lymphocytes in CD4 T lymphocytes and inhibits the proliferation of Th2 lymphocytes. This effect is mediated by activating monocytes to secrete IL-12 and activating T lymphocytes to express functional IL-12 receptors.
따라서 인체 내 인터페론-γ 생성에 의한 면역기능 향상을 통해 젊은 피부를 유지시킬 수 있음과 함께, 히스타민 생성 조절에 의한 알레르기와 같은 피부질환을 근본적으로 개선할 수 있는 제품의 개발이 절실히 요구되어 왔다.Therefore, there is an urgent need for the development of a product that can maintain the young skin through the improvement of immune function by the production of interferon-γ in the human body and fundamentally improve skin diseases such as allergy by regulating histamine production.
본 발명자들은 종래의 피부 트러블을 포함한 피부질환 치료제들이 갖는 문제점을 해결함과 동시에 면역기능 저하, 신기능 저하, 호르몬 분비 이상 등의 부작용도 없는 조성물을 개발해내기 위해 지속적인 연구를 수행하였다. 그 결과, 기존의 알레르기 반응에 탁월한 단일물질들을 혼합하여 사용한 경우 피부질환 개선과 아울러 면역기능 조절에도 효과가 있음을 확인하고 본 발명을 완성하였다.The present inventors conducted a continuous study to solve the problems of conventional skin diseases, including skin problems, and at the same time to develop a composition that does not have side effects such as decreased immune function, renal function, abnormal hormone secretion. As a result, it was confirmed that there is an effect to improve the skin disease as well as to regulate the immune function when used by mixing a single substance excellent in the conventional allergic reaction and completed the present invention.
따라서 본 발명은 단순한 피부개선이나 일시적 치료가 아닌, 인체 면역기능 조절에 의해 피부 트러블의 원인을 근본적으로 치료하는 조성물 및 그의 제조방법을 제공하는 것을 목적으로 한다.Accordingly, an object of the present invention is to provide a composition and a method for producing the same that fundamentally treat the cause of skin problems by regulating human immune function, rather than simply improving or temporarily treating the skin.
본 발명은 유효성분으로서, 당귀, 천궁, 생지황, 하수오, 황기, 감초, 길경, 포공영, 백작약, 백출, 갈근, 박하, 사인, 진피, 율무 및 건율을 혼합한 후 물로 추출하거나 각각 물로 추출한 후 혼합하여 얻은 수추출물 또는 그의 건조분말을 함유하는, 피부보호 및 피부질환 개선용 조성물에 관한 것이다.The present invention as an active ingredient, Angelica, Cheonggung, Saenghwang, Sewao, Astragalus, Licorice, Gilgyeong, Pogongyoung, Baekjak, Baekchul, Brown root, Peppermint, Sine, Dermis, Yulmu and dry ratio, and then extracted with water or extracted with water and then mixed The present invention relates to a composition for skin protection and skin disease improvement, comprising a water extract or dried powder thereof.
본 발명에서 원료생약으로 사용되는 당귀는 보혈화혈, 조경지통, 윤장통변하고; 천궁은 활혈행기, 거풍지통하고; 생지황은 청열양혈, 생진지갈하고; 하수오는 보간신, 익정혈, 통변, 해창독하고; 황기는 보기, 탁독, 배농, 지통작용이 있고; 감초는 진해, 거담, 해독, 진경, 항염, 진통, 항과민 효능이 있으며; 길경은 거담배농, 청리두목인후하고; 포공영은 청열해독, 소옹산결, 항균, 항바이러스, 항진균, 이뇨능이 있으며; 기타 성분들은 고표지한, 정천지해, 생기, 소산풍열, 조중행기, 조중지사, 청열배농, 이수소종 등의 효능 중 일부를 갖는다.Angelica used as a raw material herbal in the present invention is blood donation blood, landscaping pain, yunjangcheong; Celestial arches are vigorous; Saengjihwang is blue fever, lively; Sewage is interpolated, bleeding, stool, sea swelling; Astragalus has boil, turbid poison, drainage, and aching pain; Licorice has antitussive, expectorant, detoxifying, alveolar, anti-inflammatory, analgesic and anti-hypersensitive effects; Gil-Gyeong is an expectorant with a fermented stomach; Pogongyoung has the ability to detoxify, lysate, antibacterial, antiviral, antifungal, diuretic; Other ingredients have some of the efficacy of high-label, natural shallow sea, vitality, dissipative wind heat, mid-season, mid-heavy branch, clear heat drainage, and dihydrogen species.
본 발명에 있어서, 각 생약들은 통상의 방법에 의해 그 유효성분들을 각각 추출하거나, 생약들을 혼합한 후 한꺼번에 추출하고, 필요한 경우 이를 농축건조시켜 분말화한다. 본 발명에 있어서, 바람직하게는 각 생약들을 규정된 중량비로 혼합한 후, 물로 추출한다.In the present invention, each of the herbal medicines are extracted by the conventional method, the active ingredients, or after mixing the herbal medicines at once, and if necessary, concentrated to dry it to powder. In the present invention, preferably, each herbal medicine is mixed in a prescribed weight ratio and then extracted with water.
본 발명에 따른 조성물은 바람직하게는 약제학적으로나 식품 또는 화장용으로 허용되는 담체를 추가로 함유한다. 본 발명의 조성물에서 사용될 수 있는 약제학적으로 허용되는 담체는 약제학적 분야에서 통상적인 것으로, 예를 들면 경구투여용 제제의 경우에는 결합제, 활택제, 붕해제, 부형제, 가용화제, 안정화제 등이 있고, 국소투여용 제제의 경우에는 기제, 부형제, 윤활제, 보존제 등이 있다. 필요에 따라서는, 환제, 과립제, 정제 또는 캅셀제 형태로 제조할 수 있고, 필요에 따라, 사용시에 다른 제형으로 바꾸어 사용할 수도 있다. The composition according to the invention preferably further contains a pharmaceutically or food or cosmetically acceptable carrier. Pharmaceutically acceptable carriers that can be used in the compositions of the present invention are conventional in the pharmaceutical art, for example, in the case of oral preparations, binders, lubricants, disintegrants, excipients, solubilizers, stabilizers, etc. In the case of formulations for topical administration, there are bases, excipients, lubricants, preservatives and the like. If necessary, it may be prepared in the form of pills, granules, tablets or capsules, and, if necessary, may be used in a different dosage form when used.
본 발명에 따른 조성물의 제조 원료는 당귀, 천궁, 생지황, 하수오, 황기, 감초, 길경, 포공영, 백작약, 백출, 갈근, 박하, 사인, 진피, 율무 및 건율을 바람직하게는 2∼8 : 2∼8 : 2∼8 : 1∼6 : 3∼12 : 1∼6 : 2∼8 : 8∼16 : 2∼8: 2∼8: 1∼8 : 1∼8 : 0.5∼4 : 0.5∼4 : 1∼16 : 0.5∼20의 중량비로 포함된 것이다.The raw materials for the preparation of the composition according to the present invention are Angelica, Cheonggung, Saengjihwang, Sewao, Astragalus, Licorice, Gilgyeong, Pogongyoung, Baekjak, Baekchul, Brown Root, Peppermint, Sinus, Dermis, Yulmu and Dried Rate. 8: 2-8: 1-6: 3-12: 1-6: 2-8: 8-16: 2-8: 2-8: 1-8: 1-8: 0.5-4: 0.5-4: It is contained in the weight ratio of 1-16: 0.5-20.
상기 생약들의 조성비는 반복된 실험결과를 토대로 얻어진 것이며 그 하한치보다 낮을 경우에는 그 성분의 생리활성효과가 감소하며 그 상한치보다 높을 경우에는 타성분의 생리활성효과가 감소될 수 있어 조성물의 상승작용 및 협동작용이 저하될 우려가 있다.The composition ratio of the herbal medicines is obtained on the basis of repeated experimental results, and if lower than the lower limit, the physiological activity effect of the component may be reduced, and if higher than the upper limit, the physiological activity effect of the other components may be reduced. There is a fear that cooperation will be reduced.
본 발명의 조성물은 피부보호 및 피부질환 개선제로서 사용될 수 있으며, 이러한 효과는 하기 실험예 1 내지 4로부터 확인된다. 본 발명에 따른 조성물은 면역 관련세포로부터 사이토카인 및 화학적 매개물질의 생성을 조절하여 피부보호 및 피부질환 개선효과를 증강하는 것으로 추정된다.The composition of the present invention can be used as a skin protection and skin disease improving agent, this effect is confirmed from Experimental Examples 1 to 4 below. The composition according to the present invention is believed to enhance skin protection and skin disease improvement by regulating the production of cytokines and chemical mediators from immune-related cells.
본 발명의 조성물은 예를 들어 당귀 2∼8 g, 천궁 2∼8 g, 생지황 2∼8 g, 하수오 1∼6 g, 황기 3∼12 g, 감초 1∼6 g, 길경 2∼8 g, 포공영 8∼16 g, 백작약 2∼8 g , 백출 2∼8 g, 갈근 1∼8 g, 박하 1∼8 g, 사인 0.5∼4 g, 진피 0.5∼4 g, 율무 1∼16 g 및 건율 0.5∼20 g의 혼합물에 물 800 ㎖를 붓고 약 2 시간 동안 달여 약 150 ㎖로 농축시켜 탕제로 만든다.The composition of the present invention, for example, Angelica 2-8 g, Cheongung 2-8 g, Saengjihwang 2-8 g, Sewage 1-6 g, Astragalus 3-12 g, Licorice 1-6 g, Gil length 2-8 g, Pogongyoung 8-16 g, Baekjak 2-8 g, Baekchul 2-8 g, Brown root 1-8 g, Mint 1-8 g, Sine 0.5-4 g, Dermis 0.5-4 g, Young radish 1-16 g, and dry rate 0.5 800 ml of water are poured into a mixture of -20 g, decocted for about 2 hours, concentrated to about 150 ml and made into a tan.
본 조성물의 통상적인 1 회 투여용량은 상기 탕제 기준 체중 1 ㎏ 당 1.5∼2.5 ㎖이며 이를 1 일 3 회 복용한다. 예를 들어, 체중 60 ㎏인 성인의 경 우, 1 회 90∼150 ㎖씩 1 일 3 회 복용한다. 이 용량의 액제를 동결건조 분말로 할 경우 0.5∼1 g/1 회에 달한다. 그러나 본 조성물의 복용량은 환자의 체중, 연령, 성별, 병증의 경중정도와 소화상태에 따라 적의 증감될 수 있다. 기타 제제에 있어서도 상기 액제 복용량을 고려하여 산출된 적정량을 경구 투여한다.
A typical single dose of the composition is 1.5 to 2.5 ml per kg of body weight based on the type of tanning and is taken three times a day. For example, for an adult weighing 60 kg, take 90-150 ml once three times a day. When the solution of this capacity is used as a lyophilized powder, it is 0.5 to 1 g / 1 times. However, the dosage of the composition may be increased or decreased depending on the patient's weight, age, sex, severity of the condition and digestion. In other formulations, the appropriate amount calculated in consideration of the liquid dosage is administered orally.
이하, 본 발명을 하기 실시예에 의하여 보다 구체적으로 설명하지만 이들에 의해 본 발명의 범위가 어떤 식으로든 제한되는 것은 아니다.
Hereinafter, the present invention will be described in more detail with reference to the following Examples. The scope of the present invention is not limited in any way.
실시예 1: 액제의 제조Example 1 Preparation of Liquid
당귀 4 g, 천궁 4 g, 생지황 4 g, 하수오 3 g, 황기 6 g, 감초 3 g, 길경 4 g, 포공영 8 g, 백작약 4 g, 백출 4 g, 갈근 4 g, 박하 4 g, 사인 2 g, 진피 2 g, 율무 8 g 및 건율 10 g의 혼합물에 물 800 ㎖를 붓고 약 2 시간 동안 달여 약 150 ㎖로 농축시켜 액제로 만들었다.
Angelica 4g, 4 g of Cheongung, 4 g of raw jihwang, 4 g of sewage, 3 g of astragalus, 6 g of licorice, 3 g of licorice, 4 g of pogongyoung, ginseng 4 g, white extract 4 g, black root 4 g, mint 4 g, signature 2 To a mixture of g, 2 g of dermis, 8 g of barley radish and 10 g of dryness was poured 800 ml of water, decocted for about 2 hours and concentrated to about 150 ml to form a liquid.
실시예 2: 기타 제제의 제조Example 2: Preparation of Other Formulations
통상의 방법에 따라 실시예 1의 추출물로 환제, 과립제, 정제 및 캅셀제를 제조하였다.
Pills, granules, tablets and capsules were prepared from the extract of Example 1 according to a conventional method.
실시예 3: 식품의 제조Example 3: Preparation of Food
실시예 1의 추출물을 동결건조한 후 그 분말을 초콜렛과 혼합하여 통상의 방 법에 따라 식품으로 제조하였다.
After freeze-drying the extract of Example 1, the powder was mixed with chocolate to prepare a food according to a conventional method.
실험예 1: 본 발명에 따른 조성물의 히스타민 방출 억제효과 측정Experimental Example 1 Measurement of the Histamine Release Inhibitory Effect of the Composition According to the Present Invention
본 발명에 따른 조성물의 히스타민 방출 억제효과를 측정하기 위하여, 아래와 같은 실험을 수행하였다. In order to measure the histamine release inhibitory effect of the composition according to the present invention, the following experiment was performed.
흰쥐(mouse) 복강에 30 ㎖ 정도의 타이로드(Tyrode) 완충용액 B(137 mM NaCl, 5.3 mM 글루코스, 12 mM NaHCO3, 2.7 mM KCl, 0.3 mM NaH2PO4)를 주입하고 복부를 90 초 정도 마사지하였다. 그 후 복막을 조심스럽게 열어 파스퇴르 피펫으로 복강내 세포를 함유하고 있는 타이로드 완충용액 B를 수거하였다. 이어서 상기 세포 함유 용액을 150×g로 10 분 동안 원심분리하여 세포를 침전시켰다. 침전된 세포에 타이로드 완충용액 B를 가하여 피펫팅한 후, 22.5% 메트리자마이드(metrizamide) 용액위에 살짝 적층하고 400×g에서 15 분 동안 원심분리하여 침전된 세포만을 얻었다. 상기 침전된 세포에 α-MEM(minimum essential medium)/50% FBS(fetal bovine serum)를 가하고, 피펫팅하여 톨루이딘 블루 염색 방법을 통해 비만세포가 95% 이상이고 트립판 블루 염색에 의해 생존능력이 97% 이상임을 확인하고, 실험에 사용하였다. Inject the rat abdominal cavity with about 30 ml of Tyrode buffer B (137 mM NaCl, 5.3 mM glucose, 12 mM NaHCO 3 , 2.7 mM KCl, 0.3 mM NaH 2 PO 4 ) and inhale the abdomen for 90 seconds. Massage about. Then, the peritoneum was carefully opened to collect Tyrod buffer B containing intraperitoneal cells with a Pasteur pipette. The cell-containing solution was then centrifuged at 150 x g for 10 minutes to precipitate the cells. Tirod buffer B was added to the precipitated cells, followed by pipetting, followed by slightly stacking on 22.5% metrizamide solution and centrifugation at 400 × g for 15 minutes to obtain only precipitated cells. Α-MEM (minimum essential medium) / 50% FBS (fetal bovine serum) was added to the precipitated cells and pipetted to mast cells by 95% or more by toluidine blue staining method and viability by trypan blue staining. It was confirmed that more than 97%, it was used in the experiment.
상기 정제된 비만세포에 α-MEM/50% FBS를 가하고 조심스럽게 피펫팅한 후, 2×105 개의 세포가 되도록 분주하였다. 세포를 안정화시키기 위하여 실시예 1의 조성물을 처리하기 전에 37 ℃에서 10 분 동안 미리 배양하고, 실시예 1의 조성물 수용액을 각각 0.01, 0.1, 1.0 ㎎/㎖(증류수 1 ㎖당 실시예 1의 조성물 0.01, 0.1, 1 ㎎을 녹인 것)가 되도록 가하고 37 ℃에서 30 분 동안 처리하였다. 그 후 곧 바로 '화합물 48/80'(Baltzly et al, 1949. A family of long acting depressors. Journal of American Chemical Society 71, 1301-1305) 각각 5.0 ㎍/㎖(부피당 시약의 양)을 10 분 동안 처리하였다. 반응 종료 후 400×g에서 원심분리하여 상층액과 세포를 분리하였다.Α-MEM / 50% FBS was added to the purified mast cells and carefully pipetted, and then divided into 2 × 10 5 cells. Incubate at 37 ° C. for 10 minutes prior to treatment of the composition of Example 1 to stabilize the cells, and the aqueous solution of Example 1 was 0.01, 0.1, 1.0 mg / ml (the composition of Example 1 per ml of distilled water, respectively). 0.01, 0.1, 1 mg dissolved) and treated at 37 ° C. for 30 minutes. Shortly thereafter, 5.0 μg / ml (amount of reagent per volume) was added for 10 minutes for each compound 48/80 (Baltzly et al, 1949. A family of long acting depressors. Journal of American Chemical Society 71, 1301-1305). Treated. After completion of the reaction, the supernatant and the cells were separated by centrifugation at 400 × g.
히스타민 정량은 OPA 스펙트로플루오로메트리(O-phthaldialdehyde spectrofluorometry) 방법으로 438 ㎚에서 측정하였으며, 히스타민 분비의 억제율을 하기 식을 사용하여 계산하였다:Histamine quantitation was measured at 438 nm by OPA-phthaldialdehyde spectrofluorometry, and the inhibition rate of histamine secretion was calculated using the following formula:
억제율(%)=[(A-B)×100]/AInhibition Rate (%) = [(A-B) × 100] / A
A: 약물을 부가하지 않았을 때의 히스타민 양 A: amount of histamine when no drug is added
B: 약물을 부가하였을 때의 히스타민 양 B: amount of histamine when drug is added
그 결과를 표 1에 나타내었다.The results are shown in Table 1.
표 1에 나타낸 바와 같이, '화합물 48/80'에 의하여 유도된 복강 비만세포로부터 히스타민의 분비가 본 발명의 조성물 0.01 ㎎/㎖의 농도에서 약 20% 억제됨을 확인할 수 있었다.
As shown in Table 1, it was confirmed that the secretion of histamine from the peritoneal mast cells induced by 'Compound 48/80' was suppressed by about 20% at the concentration of 0.01 mg / ml of the composition of the present invention.
실험예 2: 본 발명에 따른 조성물의 피부 알레르기 반응에 대한 효과 측정Experimental Example 2: Determination of the effect on the skin allergic reaction of the composition according to the present invention
본 발명에 따른 조성물의 수동적 피부 알레르기 반응에 대한 효과를 조사하기 위하여, 아래와 같은 방법으로 인위적인 피부 알레르기(passive cutaneous anaphylaxis, PCA) 반응을 일으켰다. In order to investigate the effect on the passive skin allergic reaction of the composition according to the present invention, a passive cutaneous anaphylaxis (PCA) reaction was caused by the following method.
실험동물로서 4∼6 주령의 체중 5∼35 g인 ICR계 흰쥐를 실험 군별로 5 마리씩 사용하여 반복 실험하였다. 먼저 충분히 섭식시킨 후 24 시간이 경과한 상기 흰쥐를 대상으로 하였다. 상기 흰쥐는 대한 동물센터로부터 입수하였다.As an experimental animal, five ICR rats weighing 5 to 35 g at 4 to 6 weeks of age were repeatedly used in each experimental group. First, the rats were fed 24 hours after sufficient feeding. The rats were obtained from the Korean Animal Center.
항-DNP(dinitrophenyl) IgE를 상기 흰쥐의 피내에 주입하여 피부를 감작시킨 다음, 48 시간 후에 항원 DNP-HSA(dinitrophenyl-human serum albumin)를 꼬리에 정맥 주사하여 IgE 의존성 피부 반응을 유발하였다. 이때 DNP-HSA는 PBS(phosphate buffered saline)로 희석하였다. 흰쥐의 등에 항-DNP IgE 100 ng을 주입하고 48 시간 경과 후 4% 에반스 블루(evans blue)와 DNP-HSA를 1:1로 혼합한 용액을 쥐의 꼬리정맥에 주사하였다. 항원(DNP-HSA)을 투여하기 1 시간 전에 실시예 1의 추출물 0.01, 0.1 및 1 g/체중 ㎏을 흰쥐에 각각 구강 투여하였다. The skin was sensitized by injecting anti-DNP (dinitrophenyl) IgE into the skin of the rats, and then 48 hours later, the antigen DNP-HSA (dinitrophenyl-human serum albumin) was injected intravenously into the tail to induce an IgE dependent skin response. At this time, DNP-HSA was diluted with PBS (phosphate buffered saline). 100 ng of anti-DNP IgE was injected into the rat's back, and 48 hours later, a mixture of 4% Evans blue and DNP-HSA 1: 1 was injected into the tail vein of the rat. One hour before administration of the antigen (DNP-HSA), 0.01, 0.1 and 1 g / kg body weight of the extract of Example 1 were orally administered to the rats, respectively.
흰쥐를 치사시킨 후 반응이 일어난 부위를 절단하여 0.1 N 수산화칼륨 500 ㎕를 첨가한 후, 아세톤과 인산(5:13(w:w)) 혼합용액을 4.5 ㎖씩 가하여 색소를 추출하였다. 620 ㎚에서 추출물의 흡광도를 측정하였으며, 에반스 블루 표준 곡선을 이용하여 색소량을 측정하였다. 그 결과를 표 2에 나타내었다. After killing the rats, the reaction site was cut and 500 μl of 0.1 N potassium hydroxide was added. Then, 4.5 ml of acetone and phosphoric acid (5:13 (w: w)) mixed solution was added to extract pigments. The absorbance of the extract was measured at 620 nm and the amount of pigment was measured using the Evans Blue standard curve. The results are shown in Table 2.
표 2에 나타낸 바와 같이, 0.01 g/㎏ 농도의 조성물에서 수동적 피부 알레르기 반응을 가장 많이 억제하는 것으로 나타났다. 이러한 결과는 본 발명에 따른 조성물의 경구투여를 통해 국소, 즉 피부 알레르기 반응을 현저하게 억제할 수 있음을 의미한다.
As shown in Table 2, it was found that the most inhibiting passive skin allergic reaction in the composition of 0.01 g / kg concentration. This result means that the oral administration of the composition according to the present invention can significantly suppress local, that is, skin allergic reactions.
실험예 3: 본 발명에 따른 조성물의 이개 부종반응에 대한 효과 측정Experimental Example 3: Determination of the effect of the composition according to the invention on the edema reaction
실험동물로서 4∼6 주령의 체중 25∼35 g인 ICR계 흰쥐를 실험 군별로 4 마리씩 사용하여 반복 실험하였다. 먼저 충분히 섭식시킨 후 24 시간 경과한 상기 흰쥐를 대상으로 하였다. 상기 흰쥐는 대한 동물센터로부터 입수하였다.As experimental animals, four ICR rats weighing 25 to 35 g at 4 to 6 weeks of age were repeatedly used in each experimental group. First, the rats were fed 24 hours after sufficient feeding. The rats were obtained from the Korean Animal Center.
먼저 실시예 1의 조성물을 농도별로(0.01, 0.1 및 1 g/㎏) 경구 투여한 후 1 시간 동안 반응시켰다. 그 후 화합물 48/80(20 g/ℓ)을 미세 주사기를 사용하여 쥐 귀 피하 내부에 주입하고 40 분 후에 이개 부종반응이 일어난 부위의 귀 두께를 측정하였다. 그 결과를 표 3에 나타내었다. First, the composition of Example 1 was orally administered by concentration (0.01, 0.1, and 1 g / kg), and then reacted for 1 hour. Compound 48/80 (20 g / L) was then injected subcutaneously inside the rat ear using a micro syringe and the ear thickness of the site where the edema reaction occurred after 40 minutes was measured. The results are shown in Table 3.
표 3에서 나타낸 바와 같이, 본 발명에 따른 조성물은 낮은 농도에서는 유의한 효과가 없고, 1 g/㎏의 농도에서 이개 부종반응을 가장 많이 억제함을 알 수 있었으며, P<0.005로 유의한 결과를 얻었다.
As shown in Table 3, the composition according to the present invention did not have a significant effect at low concentrations, it was found that the most suppress the edema reaction at a concentration of 1 g / ㎏, P <0.005 significant results Got it.
실험예 4: 본 발명에 따른 조성물의 인터페론-γ 생성 효과 측정Experimental Example 4: Determination of interferon-γ production effect of the composition according to the present invention
본 발명에 따른 조성물이 면역증강 기능을 담당하고 있는 인터페론-γ 생성을 유도하는지 여부를 아래와 같이 조사하였다.Whether the composition according to the invention induces the production of interferon-γ responsible for the immune enhancing function was investigated as follows.
배양액 RPMI 1640/FBS에 T 세포주 Molt-4를 5% CO2, 37 ℃에서 배양한 후 3×105 개의 세포가 되도록 분주하였다. 세포를 안정화시키기 위하여 실시예 1의 조성물을 처리하기 전에 37 ℃에서 10 분 동안 미리 배양하고, 실시예 1의 조성물 수용액을 각각 0.01, 0.1, 1.0 ㎎/㎖(증류수 1 ㎖당 실시예 1의 조성물 0.01, 0.1, 1 ㎎을 녹인 것)가 되도록 가한 다음 24 시간동안 반응시켰다. 반응 종료 후 400×g에서 원심분리하여 상층액과 세포를 분리하였다. 분리된 상층액 중 인터페론-γ의 단백질 수준을 조사하기 위하여 ELISA(enzyme linked immunosorbent assay)를 아래 와 같이 실행하였다.T cell line Molt-4 was cultured in culture medium RPMI 1640 / FBS at 5% CO 2 , 37 ° C., and then aliquoted to 3 × 10 5 cells. Incubate at 37 ° C. for 10 minutes prior to treatment of the composition of Example 1 to stabilize the cells, and the aqueous solution of Example 1 was 0.01, 0.1, 1.0 mg / ml (the composition of Example 1 per ml of distilled water, respectively). 0.01, 0.1, 1 mg dissolved), and reacted for 24 hours. After completion of the reaction, the supernatant and the cells were separated by centrifugation at 400 × g. In order to investigate the protein level of interferon-γ in the separated supernatant, an enzyme linked immunosorbent assay (ELISA) was performed as follows.
인터페론-γ에 대한 단일클론 항체인 항-인간 인터페론-γ(1 ㎍/㎖)를 PBS(pH 7.4)로 희석하여, 96-웰 플레이트에 100 ㎕씩 각각 코팅한 다음, 4 ℃에서 12 시간 동안 방치하였다. 상기 플레이트를 0.05% 트윈이 포함된 PBS로 세척한 다음, 1% BSA, 5% 슈크로스, 0.05% NaN3를 함유한 PBS로 1 시간 동안 차단시켰다. 여러 차례 세척한 다음, 원심 분리하여 얻은 상층액과 표준 곡선으로 사용할 재조합 인터페론-γ를 첨가한 후 37 ℃에서 2 시간 동안 방치하였다. 이어서 각 웰을 다시 세척하고 2차 항체 바이오틴(biotin)이 결합된 인터페론-γ(0.2 ㎍/㎖)를 첨가하여 다시 37 ℃에서 2 시간 동안 방치하였다. 웰을 세척한 다음, 아비딘(avidine)이 연결된 과산화효소를 첨가하고 37 ℃에서 20 분 동안 방치하였다. 웰을 다시 세척한 다음, ABTS(2,2′-AZINO-bis(3-ethylbenzthiazoline-6-6-sulfonic acid)) 기질을 첨가하고, 발색반응을 ELISA 판독기를 사용하여 405 ㎚에서 측정하였다.Anti-human interferon-γ (1 μg / ml), a monoclonal antibody against interferon-γ, was diluted with PBS (pH 7.4) and coated in 100 μl each in a 96-well plate, followed by 12 h at 4 ° C. It was left. The plate was washed with PBS containing 0.05% Tween and then blocked for 1 hour with PBS containing 1% BSA, 5% sucrose, 0.05% NaN 3 . After washing several times, the supernatant obtained by centrifugation and recombinant interferon-γ to be used as a standard curve were added and then left at 37 ° C. for 2 hours. Each well was then washed again and added to secondary antibody biotin-bound interferon-γ (0.2 μg / ml) and left for another 2 hours at 37 ° C. The wells were washed and then avidin-linked peroxidase was added and left at 37 ° C. for 20 minutes. The wells were washed again, then ABTS (2,2′-AZINO-bis (3-ethylbenzthiazoline-6-6-sulfonic acid)) substrate was added, and color development was measured at 405 nm using an ELISA reader.
그 결과를 도 1에 나타내었다. 도 1에 나타낸 바와 같이, 본 발명에 따른 조성물 0.01 ㎎/㎖에서 인터페론-γ 생성을 가장 많이 증가시킴을 알 수 있었다(B, 무처리; *P=0.014).The results are shown in FIG. As shown in FIG. 1, it was found that the production of interferon-γ was most increased in 0.01 mg / ml of the composition according to the present invention (B, no treatment; * P = 0.014).
이상 살펴본 바와 같이, 본 발명의 조성물은 히스타민 분비를 억제하고, 수동적 피부 알레르기 반응 및 이개 부종반응 등의 알레르기 반응을 억제함으로써, 아토피 피부염과 같은 알레르기 피부질환 등의 치료제로 사용될 수 있으며, 인체 면역 기능 조절에 중요한 인터페론-γ 생성을 증가시킴으로써 면역 기능 악화 혹은 저하에 의한 노화로 인한 주름, 건선, 피부건조증, 피부가려움증, 만성 습진 및 여드름의 치료 보조제로서 효과적으로 사용될 수 있다. 또한, 복용시 인체의 피부를 매우 부드럽게 하므로, 피부미용에도 효과적인 경구용 화장품으로 사용될 수 있다.As described above, the composition of the present invention can be used as a therapeutic agent for allergic skin diseases, such as atopic dermatitis, by inhibiting histamine secretion, inhibiting allergic reactions such as passive skin allergic reaction and edema edema. By increasing interferon- [gamma] production, which is important for regulation, it can be effectively used as an adjuvant for the treatment of wrinkles, psoriasis, dry skin, itching, chronic eczema and acne due to aging due to deterioration or degradation of immune function. In addition, since the skin of the human body is very soft when taken, it can be used as an effective oral cosmetic for skin care.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101912469A (en) * | 2010-05-25 | 2010-12-15 | 中国人民解放军军事医学科学院放射与辐射医学研究所 | Application of extract of bolus for reinforcing middle jiao and replenishing qi in radiation protection |
| KR101753687B1 (en) | 2015-05-20 | 2017-07-05 | 한국과학기술연구원 | Composition for anti-allergy and skin hydration comprising fraxinus rhynchophylla extracts |
| KR20180007269A (en) * | 2016-07-12 | 2018-01-22 | 이종윤 | Method for producing an extraction with high level from Phellinus linteus Quel and it's using method. |
Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR100821631B1 (en) * | 2002-08-17 | 2008-04-16 | (주)아모레퍼시픽 | Herbal Skin Care Food Composition |
| KR100732564B1 (en) * | 2005-08-22 | 2007-06-27 | 한불화장품주식회사 | External skin composition containing dermis extract as main active ingredient |
| KR100761329B1 (en) * | 2006-04-28 | 2007-10-04 | 주식회사 장생도라지 | Composition for external application of skin containing Jangsaeng Doraji extract and purified fractions isolated from it, showing anti-aging and anti-wrinkle formation |
| WO2015182799A1 (en) * | 2014-05-27 | 2015-12-03 | 윤종성 | Composition for improving skin diseases comprising hovenia dulcis and semen allii tuberosi |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR19980086246A (en) * | 1997-05-31 | 1998-12-05 | 김인규 | Extraction method of hair growth promoting ingredient from herbal medicine and extract thereof |
| KR20010068273A (en) * | 2000-01-03 | 2001-07-23 | 김형민 | Pharmaceutical compositions for preventing and treating allergic diseases and methods for preparation thereof |
| KR20010100551A (en) * | 2000-05-03 | 2001-11-14 | 장인순 | The pharmaceutical composition and its preparation method of herb mixture for immunomodulation, heamatopoiesis augmentation and protection from radiation |
| KR20030008080A (en) * | 2001-07-16 | 2003-01-24 | 김희숙 | A composition with a good immunostimulating activity and a functional food comprising the same |
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2002
- 2002-07-19 KR KR1020020042596A patent/KR100881445B1/en not_active Expired - Fee Related
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR19980086246A (en) * | 1997-05-31 | 1998-12-05 | 김인규 | Extraction method of hair growth promoting ingredient from herbal medicine and extract thereof |
| KR20010068273A (en) * | 2000-01-03 | 2001-07-23 | 김형민 | Pharmaceutical compositions for preventing and treating allergic diseases and methods for preparation thereof |
| KR20010100551A (en) * | 2000-05-03 | 2001-11-14 | 장인순 | The pharmaceutical composition and its preparation method of herb mixture for immunomodulation, heamatopoiesis augmentation and protection from radiation |
| KR20030008080A (en) * | 2001-07-16 | 2003-01-24 | 김희숙 | A composition with a good immunostimulating activity and a functional food comprising the same |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101912469A (en) * | 2010-05-25 | 2010-12-15 | 中国人民解放军军事医学科学院放射与辐射医学研究所 | Application of extract of bolus for reinforcing middle jiao and replenishing qi in radiation protection |
| KR101753687B1 (en) | 2015-05-20 | 2017-07-05 | 한국과학기술연구원 | Composition for anti-allergy and skin hydration comprising fraxinus rhynchophylla extracts |
| KR20180007269A (en) * | 2016-07-12 | 2018-01-22 | 이종윤 | Method for producing an extraction with high level from Phellinus linteus Quel and it's using method. |
| KR101893394B1 (en) * | 2016-07-12 | 2018-08-30 | 이종윤 | Method for producing an extraction with high level from Phellinus linteus Quel and it's using method. |
Also Published As
| Publication number | Publication date |
|---|---|
| KR20040008871A (en) | 2004-01-31 |
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