KR20190051150A - Antiallergic composition comprising Allomyrina dichotoma larva extract - Google Patents
Antiallergic composition comprising Allomyrina dichotoma larva extract Download PDFInfo
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- KR20190051150A KR20190051150A KR1020170146484A KR20170146484A KR20190051150A KR 20190051150 A KR20190051150 A KR 20190051150A KR 1020170146484 A KR1020170146484 A KR 1020170146484A KR 20170146484 A KR20170146484 A KR 20170146484A KR 20190051150 A KR20190051150 A KR 20190051150A
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- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
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Abstract
Description
본 발명은 장수풍뎅이 유충의 추출물을 함유하는 항알레르기성 조성물에 관한 것으로, 더욱 상세하게는 β-헥소사미노다아제와 같은 알레르기 질환에서 유발되는 인자의 분비를 억제하여 아나필락시스 쇼크, 식품 알레르기, 알레르기성 비염, 화분증, 천식, 약제 알레르기, 식물 알레르기, 두드러기, 습진 및 알레르기성 접촉성 피부염 등과 같은 각종 알레르기 질환의 치료용 조성물이나 건강기능식품으로 제공될 수 있는 장수풍뎅이 유충의 추출물을 함유하는 항알레르기성 조성물에 관한 것이다.The present invention relates to an antiallergic composition containing an extract of a long-beetle beet larva, and more particularly to an antiallergic composition containing an extract of a long-beetle beet larva, and more particularly to an antiallergic composition which suppresses secretion of factors induced in an allergic disease such as beta -hexosaminidase to produce anaphylactic shock, A composition for the treatment of various allergic diseases such as asthma, rhinitis, hay fever, asthma, drug allergy, plant allergies, urticaria, eczema and allergic contact dermatitis, or an antiallergic ≪ / RTI >
경제가 성장함에 따라 생활수준이 향상되고, 서구적인 식생활의 변화 등 환경적인 원인으로 인하여 우리나라뿐만 아니라 전 세계적으로 발병률이 증가되고 있는 천식은 알레르기 반응으로부터 유발 되는 것으로 보고되고 있으며, 개인의 차가 매우 크게 나타나는 질병이다[Clin. Exp. Allergy 36, 122-132]. 천식의 주요 원인인 알레르기(allergy)란 일반적으로 꽃가루, 곰팡이, 집 먼지 진드기, 난 알부민, 동물비듬 등 여러 가지 알레르겐(allergen)에 의해 신체내의 면역학적 기전을 통해 야기되는 반응으로서 피부, 호흡기, 위장관의 점막, 임파관 주위, 혈관 주위, 뇌 등 전신의 장기에 널리 분포 하고 있는 비만세포에 의해 야기된다[Arch. Pharm. Res. 22, 320-323]. 비만세포는 알레르기 반응을 매개하는 중요한 세포이며, 천식과 알레르기성 비염에 관여하고[Crit. Rev. Immunol. 3, 23-74], 알레르겐과 결합을 통해 활성화 되면 히스타민, β-hexosaminidase, prostaglandin (PG), 류코트리엔과 같은 화학매체를 탈과립 과정을 통하여 세포외로 분비한다[Ann. N. Y. Acad. Sci. 714, 136-42]. 알레르기 반응을 확인하기 위해서는 주로 비만세포에서 분비되는 β-hexosaminidase를 측정 하는데, 이는 비만세포에서 탈과립을 통하여 분비되는 히스타민의 농도가 매우 낮기 때문에 호염구나 비만세포에 다량 존재하는 β-hexosaminidase를 천식 및 비염과 같은 알레르기 질환에서 탈과립의 지표로 이용되며, 알레르기 억제물질의 생리활성 측정에 유용하게 사용되고 있다[Arch. Pharm. Res. 22, 320-323; Korea J. Food Nutr. 15, 382-385]. As the economy grows, the level of living improves, and asthma, which is increasing worldwide as well as in Korea due to environmental factors such as changes in Western dietary habits, is reported to be caused by allergic reactions. It is an emerging disease [ Clin. Exp. Allergy 36,122-132]. Allergy, a major cause of asthma, is a reaction caused by various immunological mechanisms in the body caused by various allergens such as pollen, mold, house dust mites, egg albumin, and animal dander, Mucous membranes, around the lymphatic vessels, around the blood vessels, and in the organs of the whole body such as the brain [ Arch. Pharm. Res. 22, 320-323]. Mast cells are important cells that mediate allergic reactions, and are involved in asthma and allergic rhinitis [ Crit. Rev. Immunol. 3, 23-74]. When activated through binding with allergens, chemical media such as histamine, β-hexosaminidase, prostaglandin (PG) and leukotrienes are secreted out of the cell via degranulation [ Ann. NY Acad. Sci. 714, 136-42]. In order to identify allergic reactions, mainly β-hexosaminidase secreted from mast cells is measured. This is because the concentration of histamine secreted through degranulation in mast cells is very low, so β-hexosaminidase, which is abundant in basophils and mast cells, Is used as an indicator of degranulation in allergic diseases such as Alzheimer's disease, and is useful for measuring the physiological activities of allergy-suppressing substances ( Arch. Pharm. Res. 22, 320-323; Korea J. Food Nutr. 15,382-385.
이와 같이 알레르기 반응은 비만세포의 탈과립 등 비만세포의 활성화에 의해 야기되므로, 비만세포의 활성화를 억제하는 물질은 알레르기성 질환의 치료제로 활용될 수 있다[Appl. Biol. Chem. 48, 315-321]. 현재 알레르기 질환과 염증의 치료 방법은 염증 억제제인 스테로이드제 등의 외용제나 연고, 항히스타민제의 도포나 복용, 면역반응 억제제가 사용되고 있다[J. Korean Soc. Food Sci. Nutr. 42, 1378-1386]. 그러나 이러한 치료제는 증상을 완화하는데 매우 효과적이나 장기간 사용 시 부작용 및 합병증이 있으며 약 복용을 중단하면 병변의 재발 가능성이 높아지므로 안전하고 효과적인 대체 치료법이 요구된다[Dermatol. Clin. 18, 1-11; Immunol. Today 13, 136-42]. 현재까지 천연물을 기반으로 하는 천식치료제 개발은 주로 식물을 대상으로 많은 연구가 수행되어 왔으나, 아직 합성의약을 대체할 수 있는 새로운 천연물 치료제 개발 실적은 미흡한 실정이다. Since the allergic reaction is caused by activation of mast cells such as degranulation of mast cells, a substance that inhibits the activation of mast cells can be used as a therapeutic agent for allergic diseases [ Appl. Biol. Chem. 48, 315-321]. Currently, methods for treating allergic diseases and inflammation include external agents such as steroids, ointments, application or administration of antihistamines, and immunosuppressive agents, which are antiinflammatory agents [ J. Korean Soc. Food Sci. Nutr. 42,1378-1386). However, these treatments are very effective in alleviating symptoms, but there are side effects and complications in long-term use, and the possibility of recurrence of lesions after discontinuation of medication is high, thus requiring safe and effective alternative therapies [ Dermatol. Clin. 18, 1-11; Immunol. Today 13, 136-42]. To date, many studies have been conducted on plants for the development of asthma medicines based on natural products, but the development of new natural remedies that can replace synthetic medicines has not yet been completed.
곤충들은 살아가는 환경에 적응하고 생존하기 위해서 그들만의 특별한 생리조절 기전 및 물질들을 확보하는 방향으로 진화해 왔는데, 최근 곤충의 생리활성 물질들을 이용한 새로운 약물후보물질 개발 연구가 활발히 진행되고 있다[Insect Biochem. Mol. Biol. 41, 747-769]. Insects have evolved to acquire their unique physiological regulatory mechanisms and substances in order to adapt to the living environment and to survive. Recently, research on the development of novel drug candidates using insect bioactive materials has been actively conducted [ Insect Biochem . Mol. Biol. 41, 747-769.
장수풍뎅이(Allomyrina dichotoma)는 딱정벌레목 장수풍뎅이과의 곤충으로 한국, 일본, 중국, 인도에 분포하고 있으며, 상수리나무 또는 졸참나무와 같은 여러 종류의 나무에 서식하는 것으로 보고되어있다. 특히 장수풍뎅이 유충(Allomyrina dichotoma larva ; AD)은 전통적으로 간질환을 치료하기 위한 민간요법으로 사용되어 왔을 뿐만 아니라[J. Exp. Clin. Cancer. Res. 28, 112; J. Ethnopharmacol. 65, 207-216], 필수불포화지방산 및 단백질의 함량이 높은 것으로 알려져 건강식품으로써의 가능성이 보고 된 바가 있다[Korea J. Herbol. 26, 1-7]. 또한 최근 생화학적인 생리활성 연구에 따르면 장수풍뎅이 유충은 활성산소를 억제하는 항산화 효능[J. Photochem. Photobiol. B. 99, 67-73] 또는 항비만[Entomol. Res. 44, 9-16], 신경세포보호[Entomol. Res. 44, 23-30] 효과에도 탁월한 기능을 가지고 있는 것으로 보고되고 있다. Allomyrina dichotoma is an insect species of the beetle, and is distributed in Korea, Japan, China and India. It is reported to live in various kinds of trees such as oak tree or quercus. In particular, the Allomyrina dichotoma larva (AD) has traditionally been used as a folk remedy to treat liver disease [ J. Exp. Clin. Cancer. Res. 28, 112; J. Ethnopharmacol. 65, 207-216], the content of essential unsaturated fatty acids and proteins is known to be high, and the possibility as a health food has been reported [ Korea J. Herbol. 26, 1-7]. Recent biochemical physiological activity studies have also shown that long-lived beetle larvae have antioxidant potency to inhibit reactive oxygen species [ J. Photochem. Photobiol. B. 99, 67-73] or anti-obesity [ Entomol. Res. 44, 9-16], neuronal cell protection [ Entomol. Res. 44, 23-30] have been reported to have excellent functions.
이에 본 발명자들은 장수풍뎅이(Allomyrina dichotoma) 유래의 추출물을 이용하여 다양한 생리활성을 연구하던 중, 상기 추출물이 항알레르기성 효능이 우수함을 확인함으로써 본 발명을 완성할 수 있었다. Accordingly, the present inventors have been able to complete the present invention by studying various physiological activities using an extract derived from an Allomyrina dichotoma , confirming that the above extract has excellent antiallergic efficacy.
본 발명의 목적은 장수풍뎅이 유충의 추출물을 함유하는 항알레르기성 조성물을 제공하는데 있으며, 보다 자세하게는 β-헥소사미노다아제, IL-4, COX-2 또는 TNF-α와 같은 알레르기 질환에서 유발되는 인자의 분비를 억제하여 아나필락시스 쇼크, 식품 알레르기, 알레르기성 비염, 화분증, 천식, 약제 알레르기, 식물 알레르기, 두드러기, 습진 및 알레르기성 접촉성 피부염 등과 같은 각종 알레르기 질환의 치료용 조성물이나 건강기능식품으로 제공될 수 있는 장수풍뎅이 유충의 추출물을 함유하는 항알레르기성 조성물을 제공하는 데에 있다.It is an object of the present invention to provide an antiallergic composition containing an extract of Longevity Beetle larvae, and more particularly to an antiallergic composition containing an extract of Longevity beetle larvae which is induced in allergic diseases such as beta -hexosaminidase, IL-4, COX-2 or TNF- It is provided as a composition or health functional food for the treatment of various allergic diseases such as anaphylactic shock, food allergy, allergic rhinitis, hay fever, asthma, drug allergy, plant allergy, urticaria, eczema and allergic contact dermatitis An allergenic composition containing an extract of Longevity Beetle larvae which can be used as an antiallergic composition.
본 발명은 장수풍뎅이 유충의 추출물을 함유하는 항알레르기성 조성물에 관한 것이다. The present invention relates to an antiallergic composition containing an extract of Longevity beetle larvae.
상기 추출물은 장수풍뎅이 유충의 열수 추출물인 것을 특징으로 한다. The extract is characterized by being a hot-water extract of long-lived beetle larvae.
상기 추출물은 β-헥소사미노다아제(β-hexosaminidase), IL-4(interleukin-4), COX-2(Cyclooxygenase-2) 또는 TNF-α(Tumor necrosis factor-α)의 분비를 억제하는 효능이 있다. The above extract has an efficacy of suppressing the secretion of? -Hexosaminidase, IL-4 (interleukin-4), COX-2 (cyclooxygenase-2) or TNF-α (Tumor necrosis factor- have.
이에 본 발명은 상기 조성물을 함유하는 알레르기 질환의 예방 또는 치료용 약학 조성물을 제공한다. Accordingly, the present invention provides a pharmaceutical composition for preventing or treating an allergic disease containing the composition.
상기 알레르기 질환은 아나필락시스 쇼크, 식품 알레르기, 알레르기성 비염, 화분증, 천식, 약제 알레르기, 식물 알레르기, 두드러기, 습진 및 알레르기성 접촉성 피부염에서 선택되는 것일 수 있다. The allergic disease may be selected from anaphylactic shock, food allergies, allergic rhinitis, hay fever, asthma, drug allergies, plant allergies, urticaria, eczema and allergic contact dermatitis.
본 발명은 또한 상기 조성물을 함유하는 알레르기 질환의 예방 또는 개선용 건강기능식품에 관한 것이다. The present invention also relates to a health functional food for preventing or improving an allergic disease containing the composition.
상기 건강기능식품은 각종 드링크제, 육류, 소세지, 빵, 캔디류, 스넥류, 면류, 아이스크림, 유제품, 스프, 이온음료, 음료수, 알코올 음료, 껌, 차 및 비타민 복합제로 이루어진 군에서 선택되는 것일 수 있다. The health functional food may be selected from the group consisting of various drinks, meat, sausage, bread, candy, snack, noodle, ice cream, dairy product, soup, ionic drink, beverage, alcoholic beverage, gum, tea and vitamin complex.
이하, 본 발명을 상세히 설명한다.Hereinafter, the present invention will be described in detail.
본 발명은 장수풍뎅이 유충의 추출물을 함유하는 항알레르기성 조성물에 관한 것으로서, 상기 추출물은 장수풍뎅이 유충의 열수 추출물이며, 바람직하게는 장수풍뎅이 유충 중량의 5~20배 중량의 50~90℃ 열수를 장수풍뎅이 유충에 12~48시간 동안 가하여 얻은 액상 또는 이의 농축물인 것을 특징으로 한다. 상기 농축물은 분무건조, 감압건조, 가열건조, 열풍건조 또는 동결건조하여 분말상태로 제조할 수 있다. The present invention relates to an antiallergic composition containing an extract of a long beetle larva, wherein the above extract is a hot-water extract of a larva beet larva, preferably 50 to 90 ° C, Which is obtained by adding the extract to a long beetle larva for 12 to 48 hours, or a concentrate thereof. The concentrate may be prepared into a powder state by spray drying, vacuum drying, heat drying, hot air drying or lyophilization.
이렇게 하여 얻은 장수풍뎅이 유충의 열수 추출물은 에탄올이나 이의 수용액, 기타 다른 알코올류의 용매로 얻은 추출물에 비해 항알레르기 효과가 우수하다. 또한 추출 전의 시료를 바로 이용하는 것에 비해서도 수용성 성분을 유효성분으로 하는 추출물(농축물)을 이용하는 것이 항알레르기 효과가 더 뛰어나다. 또한 상기 추출물은 낮은 농도로 처리되어도 탁월한 항알레르기 효능을 나타낸다. The hot water extract of the larval beetle larvae thus obtained is superior to the extract obtained from ethanol, its aqueous solution and other alcoholic solvents. In addition, the use of an extract (concentrate) containing a water-soluble component as an active ingredient is more effective than an immediate allergy effect, as opposed to a sample immediately before extraction. The extract also exhibits excellent antiallergic efficacy even at low concentrations.
이 때 장수풍뎅이 유충은 건조분말 상태인 것을 사용할 수 있으며, 바람직하게는 장수풍뎅이 유충을 1~3일간 굶겨 배변이 완료된 상태에서 115~121℃의 온도로 고온고압 멸균한 것을 건조시킨 것일 수 있다. 이 때 유충을 굶기기 전에 급이된 장수풍뎅이 유충의 사료로는 보다 위생적인 상태의 시료를 준비하기 위해 멸균 사료를 공급하는 것이 바람직하다. In this case, the long-beetle beet larva may be in the form of a dry powder. Preferably, the long-beetle beetle larva is starved at high temperature and high pressure sterilized at a temperature of 115 to 121 ° C in a state where starving is completed for 1 to 3 days. At this time, it is preferable to feed the sterilized feed to prepare a sample of a more hygienic condition as the feed of the long-lived beetle larva fed before starving the larva.
상기 장수풍뎅이 유충의 열수 추출물은 n-헥산, 메틸렌클로라이드, 아세톤, 클로로포름, 에틸아세테이트 및 n-부탄올로 이루어진 군 중에서 선택되는 1종 이상의 용매를 사용하여 추가적으로 분획하여 분획물로 제조할 수 있다. 또는 분획용 용매로서 에테르나 아세톤 등의 기타 유기용매를 이용할 수 있다. 본 발명에서 제조한 장수풍뎅이 유충의 추출물 또는 분획물은 칼럼크로마토그래피를 이용하여 정제하여 사용할 수 있다. 상기 크로마토그래피는 실리카겔 컬럼 크로마토그래피(silica gel column chromatography), 엘에이취-20 컬럼 크로마토그래피(LH-20 column chromatography), 이온교환수지 크로마토그래피(ion exchange resin chromatography), 중압 액체 크로마토그래피(medium pressure liquid chromatography), 박층 크로마토그래피(TLC; thin layer chromatography), 실리카겔 진공 액체 크로마토그래피(silica gel vacuum liquid chromatography) 및 고성능 액체 크로마토그래피(high performance liquid chromatography) 중에서 선택될 수 있다. 본 발명에서 사용할 수 있는 추출용 기기로는 통상의 추출기기, 초음파분쇄추출기 또는 분획기를 이용할 수 있다. The hot-water extract of the long-bred beetle larvae may be further fractionated using at least one solvent selected from the group consisting of n-hexane, methylene chloride, acetone, chloroform, ethyl acetate and n-butanol. Or other organic solvents such as ether or acetone may be used as a solvent for fractionation. The extract or fraction of the larval beetle larvae prepared in the present invention can be purified and used by using column chromatography. The chromatography can be carried out using silica gel column chromatography, LH-20 column chromatography, ion exchange resin chromatography, medium pressure liquid chromatography chromatography, thin layer chromatography (TLC), silica gel vacuum liquid chromatography, and high performance liquid chromatography. As the extraction apparatus usable in the present invention, a conventional extraction apparatus, an ultrasonic pulverization extractor or a fractionator may be used.
상기 장수풍뎅이 유충의 추출물은 알레르기 질환에서 대표적으로 발현하는 단백질인 β-헥소사미노다아제(β-hexosaminidase), IL-4(interleukin-4), COX-2(Cyclooxygenase-2) 또는 TNF-α(Tumor necrosis factor-α)의 분비를 억제하는 효능이 있는 것을 특징으로 한다. 따라서 본 발명의 장수풍뎅이 유충의 추출물은 항알레르기성 조성물로 유용하게 사용될 수 있다.The extract of the long-bred beetle larvae can be used as a representative protein expressed in allergic diseases such as β-hexosaminidase, IL-4 (interleukin-4), COX-2 (cyclooxygenase- Tumor necrosis factor-alpha). Therefore, the extract of Longevity beetle larvae of the present invention can be usefully used as an antiallergic composition.
본 발명은 장수풍뎅이 유충의 추출물을 함유하는 알레르기 질환의 예방 또는 치료용 약학 조성물을 제공한다. The present invention provides a pharmaceutical composition for the prevention or treatment of an allergic disease containing an extract of Longevity Beetle larvae.
상기 알레르기 질환은 아나필락시스 쇼크, 식품 알레르기, 알레르기성 비염, 화분증, 천식, 약제 알레르기, 식물 알레르기, 두드러기, 습진 및 알레르기성 접촉성 피부염에서 선택되는 질환인 것을 특징으로 한다. The allergic disease is characterized by being selected from anaphylactic shock, food allergy, allergic rhinitis, hay fever, asthma, drug allergy, plant allergies, urticaria, eczema and allergic contact dermatitis.
상기 약학 조성물은, 각각 통상의 방법에 따라 산제, 과립제, 정제, 캡슐제, 현탁액, 에멀젼, 시럽, 에어로졸 등의 경구형 제형, 외용제, 좌제 및 멸균 주사용액의 형태로 제형화하여 사용될 수 있다. 상기 약학 조성물에 포함될 수 있는 담체, 부형제 및 희석제로는 락토즈, 덱스트로즈, 수크로스, 솔비톨, 만니톨, 자일리톨, 에리스리톨, 말티톨, 전분, 아카시아 고무, 알지네이트, 젤라틴, 칼슘 포스페이트, 칼슘 실리케이트, 셀룰로즈, 메틸 셀룰로즈, 미정질 셀룰로스, 폴리비닐 피롤리돈, 물, 메틸히드록시벤조에이트, 프로필히드록시벤조에이트, 탈크, 마그네슘 스테아레이트 및 광물유를 들 수 있다. 제제화할 경우에는 보통 사용하는 충진제, 증량제, 결합제, 습윤제, 붕해제, 계면활성제 등의 희석제 또는 부형제를 사용하여 조제된다. 경구투여를 위한 고형제제에는 정제, 환제, 산제, 과립제, 캡슐제 등이 포함되며, 이러한 고형제제는 본 발명의 추출물에 적어도 하나 이상의 부형제, 예를 들면, 전분, 탄산칼슘, 수크로스 또는 락토오스, 젤라틴 등을 섞어 조제된다. 또한 단순한 부형제 이외에 마그네슘 스테아레이트, 탈크 같은 윤활제들도 사용된다. 경구를 위한 액상 제제로는 현탁제, 내용액제, 유제, 시럽제 등이 해당되는데 흔히 사용되는 단순희석제인 물, 리퀴드 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, 좌제가 포함된다. 비수성용제, 현탁제로는 프로필렌글리콜, 폴리에틸렌글리콜, 올리브 오일과 같은 식물성 기름, 에틸올레이트와 같은 주사 가능한 에스테르 등이 사용될 수 있다. 좌제의 기제로는 위텝솔(witepsol), 마크로골, 트윈(tween) 61, 카카오지, 라우린지, 글리세로제라틴 등이 사용될 수 있다. The pharmaceutical compositions may be formulated in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols and the like, oral preparations, suppositories and sterilized injection solutions according to conventional methods. Examples of carriers, excipients and diluents that can be contained in the pharmaceutical composition include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose , Methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. In the case of formulation, a diluent or excipient such as a filler, an extender, a binder, a wetting agent, a disintegrant, or a surfactant is usually used. Solid formulations for oral administration include tablets, pills, powders, granules, capsules and the like, which may contain at least one excipient, such as starch, calcium carbonate, sucrose or lactose, Gelatin and the like. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Examples of the liquid preparation for oral use include suspensions, solutions, emulsions, and syrups. In addition to water and liquid paraffin, simple diluents commonly used, various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included . Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Examples of the suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like. Examples of the suppository base include witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatin and the like.
본 발명의 약학 조성물의 투여량은 치료받을 대상의 연령, 성별, 체중과, 치료할 특정 질환 또는 병리 상태, 질환 또는 병리 상태의 심각도, 투여경로 및 처방자의 판단에 따라 달라질 것이다. 이러한 인자에 기초한 투여량 결정은 당업자의 수준 내에 있으며, 일반적으로 투여량은 0.01㎎/㎏/일 내지 대략 2000㎎/㎏/일의 범위이다. 더 바람직한 투여량은 1㎎/㎏/일 내지 500㎎/㎏/일이다. 투여는 하루에 한번 투여할 수도 있고, 수회 나누어 투여할 수도 있다. 상기 투여량은 어떠한 면으로든 본 발명의 범위를 한정하는 것은 아니다. The dosage of the pharmaceutical composition of the present invention will depend on the age, sex, body weight of the subject to be treated, the particular disease or condition to be treated, the severity of the disease or condition, the route of administration and the judgment of the prescriber. Dosage determinations based on these factors are within the level of ordinary skill in the art and generally the dosage ranges from 0.01 mg / kg / day to approximately 2000 mg / kg / day. A more preferable dosage is 1 mg / kg / day to 500 mg / kg / day. The administration may be carried out once a day or divided into several times. The dose is not intended to limit the scope of the invention in any way.
본 발명의 약학 조성물은 쥐, 가축, 인간 등의 포유동물에 다양한 경로로 투여될 수 있다. 투여의 모든 방식은 예상될 수 있는데, 예를 들면, 경구, 직장 또는 정맥, 근육, 피하, 자궁내 경막 또는 뇌혈관내 주사에 의해 투여될 수 있다. 본 발명의 추출물은 독성 및 부작용이 거의 없으므로 예방 목적으로 장기간 복용시에도 안심하고 사용할 수 있는 약제이다. The pharmaceutical composition of the present invention can be administered to mammals such as rats, livestock, humans, and the like in various routes. All modes of administration may be expected, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intra-uterine dural or intracerebral injection. Since the extract of the present invention has little toxicity and side effects, it can be safely used even for long-term administration for the purpose of prevention.
본 발명은 상기 장수풍뎅이 유충의 추출물을 유효성분으로 포함하는 알레르기 질환의 예방 또는 개선용 건강기능식품을 제공할 수 있다. The present invention can provide a health functional food for preventing or ameliorating an allergic disease comprising the extract of Longevity beetle larvae as an effective ingredient.
상기 건강기능식품에는 식품학적으로 허용 가능한 식품보조 첨가제가 포함될 수 있다. 본 발명의 건강기능식품은 정제, 캡슐제, 환제 또는 액제 등의 형태를 포함하며, 본 발명의 추출물을 첨가할 수 있는 식품으로는, 예를 들어, 각종 드링크제, 육류, 소세지, 빵, 캔디류, 스넥류, 면류, 아이스크림, 유제품, 스프, 이온음료, 음료수, 알코올 음료, 껌, 차 및 비타민 복합제 등이 있다. The health functional food may include a food-acceptable food-aid additive. The health functional food of the present invention includes forms such as tablets, capsules, pills, and liquids. Examples of the foods to which the extract of the present invention can be added include various kinds of drinks, meat, sausage, bread, Snacks, noodles, ice cream, dairy products, soups, ionic drinks, beverages, alcoholic beverages, gums, tea and vitamin complexes.
본 발명은 장수풍뎅이 유충의 추출물을 함유하는 항알레르기성 조성물에 관한 것이다. 상기 장수풍뎅이 유충의 추출물은 β-헥소사미노다아제, IL-4, COX-2 또는 TNF-α와 같은 알레르기 질환에서 유발되는 인자의 분비를 억제하여 아나필락시스 쇼크, 식품 알레르기, 알레르기성 비염, 화분증, 천식, 약제 알레르기, 식물 알레르기, 두드러기, 습진 및 알레르기성 접촉성 피부염 등과 같은 각종 알레르기 질환의 치료용 조성물이나 건강기능식품으로 제공될 수 있다. 한편, 장수풍뎅이 유충의 추출물과 관련하여 현재까지 항알레르기성 조성물에 대한 연구는 아직까지 수행된 바 없다. 한국등록특허 제10-1686420호와 한국등록특허 제10-1480158호에는 장수풍뎅이 유충의 추출물이 염증 증상이나 류마티스 관절염의 치료 효과가 있음을 개시하고 있지만 본 발명에서처럼 장수풍뎅이 유충의 추출물이 항알레르기 효능이 있음은 확인할 수 없다. The present invention relates to an antiallergic composition containing an extract of Longevity beetle larvae. The extract of the long-bred beetle larva suppresses the secretion of factors induced in allergic diseases such as? -Hexosaminidase, IL-4, COX-2 or TNF-α and is effective as anaphylactic shock, food allergy, allergic rhinitis, Asthma, drug allergies, plant allergies, urticaria, eczema and allergic contact dermatitis, and the like. On the other hand, studies on antiallergic compositions to date have not yet been conducted in connection with the extracts of the larval beetle larvae. Korean Patent No. 10-1686420 and Korean Patent No. 10-1480158 disclose that the extract of Longevity beetle larvae has a therapeutic effect on inflammation symptoms and rheumatoid arthritis. However, as in the present invention, the extract of Longevity beetle larvae has an antiallergic effect Can not be confirmed.
도 1은 본 발명의 장수풍뎅이 유충의 추출물의 β-hexosaminidase 분비 감소 효과를 나타내는 그래프이다.
도 2는 본 발명의 장수풍뎅이 유충의 추출물의 IL-4 및 TNF-α 분비 억제 효과를 나타내는 그래프이다.
도 3은 본 발명의 장수풍뎅이 유충의 추출물의 COX-2 분비 억제 효과를 나타내는 그래프이다. 1 is a graph showing the effect of the extract of Longevity Beetle larvae of the present invention on the secretion of β-hexosaminidase.
2 is a graph showing the IL-4 and TNF-α secretion inhibitory effects of the extract of the long-beetle beetle of the present invention.
3 is a graph showing the COX-2 secretion inhibitory effect of the extract of the long-lived beetle larva of the present invention.
이하, 실시예를 통하여 본 발명을 더욱 상세히 설명하고자 한다. 이들 실시예는 오로지 본 발명을 예시하기 위한 것으로, 본 발명의 범위가 이들 실시예에 의해 제한되지 않음은 당업계에서 통상의 지식을 가진 자에게 있어서 자명할 것이다. Hereinafter, the present invention will be described in more detail with reference to Examples. It will be apparent to those skilled in the art that these embodiments are for illustrative purposes only and that the scope of the invention is not limited by these examples.
<실시예 1. 장수풍뎅이 유충의 추출물 제조>≪ Example 1: Preparation of an extract of Longevity beet larvae &
장수풍뎅이 유충(Allomyrina dichotoma larva)은 경상북도 예천군의 사육농가 '예천곤충나라'에서 구입하여 실험에 사용하였다. 장수풍뎅이 유충은 액체질소를 사용하여 급속 냉동 시킨 후 분쇄하여 분말로 제조하였으며 제조한 분말은 -70℃에서 냉동 보관하여 실험에 사용하였다. 시료의 추출은 장수풍뎅이 유충 분말 1 g에 10 ㎖의 distilled water (DW)를 혼합하여 60℃에서 24시간 교반하는 방식으로 유효 성분을 추출하였다. 이를 여과한 다음 감압농축기(rotary vaccum evaporator, CVC-3100, Tokyo, Japan)로 농축하였고 100% DMSO에 녹여 사용하였다. Allomyrina dichotoma larva was purchased from the Yecheon insect country of Yeongcheon, Gyeongbuk province and used for the experiment. The longevity of beetle larvae was rapidly frozen using liquid nitrogen and then ground into powder. The powder was stored at -70 ℃ for freezing. To extract the sample, 10 g of distilled water (DW) was mixed with 1 g of long-bean beetle larvae powder and stirred at 60 ° C for 24 hours to extract the active ingredient. After filtration, the solution was concentrated using a rotary vacuum evaporator (CVC-3100, Tokyo, Japan) and dissolved in 100% DMSO.
<실시예 2. 세포배양>≪ Example 2: Cell culture >
한국 세포주 은행으로부터 분양받은 rat basophilic leukemia 세포주인 RBL-2H3 세포를 Dulbecco's modified eagle medium (DMEM) 배지에 10% heat-inactivated fetal bovine serum (FBS)와 1% penicillin-streptomycin을 첨가하여 5% CO2 조건하에서 37℃로 유지되는 배양기에서 배양하여 사용하였다. RBL-2H3 세포는 항알레르기 관련 실험용 세포로 잘 알려져 있다[J Nutr Biochem. 48, 103-111]. RBL-2H3 cells were cultured in DMEM medium supplemented with 10% heat-inactivated fetal bovine serum (FBS) and 1% penicillin-streptomycin in 5% CO 2 Lt; RTI ID = 0.0 > 37 C. < / RTI > RBL-2H3 cells are well known as antiallergic laboratory cells [ J Nutr Biochem . 48, 103-111.
<실시예 3. β-hexosaminidase 측정>≪ Example 3: Measurement of? -Hexosaminidase >
증류수를 이용하여 추출된 장수풍뎅이 유충의 추출물의 비만세포(RBL-2H3)에 대한 세포독성을 확인하기 위해 MTS 분석을 통한 세포생존율을 조사한 결과 500 ㎍/㎖의 농도까지 세포 독성이 없는 것으로 확인되었다. The cell viability of mast cell extract (RBL-2H3) extracted with distilled water was examined by MTS analysis to confirm cytotoxicity to 500 ㎍ / ㎖ concentration .
따라서 본 발명에서의 항알레르기 효능 검증을 위한 추출물의 사용 농도는 500 ㎍/㎖을 최고 농도로 사용하였다. Therefore, the concentration of the extract used for the assay of the antiallergic activity in the present invention was 500 μg / ml at the highest concentration.
세포가 외부의 자극을 받게 되면 다양한 염증성 물질을 분비하게 되는데 그 중 비만세포는 세포 내 과립에 존재하는 다양한 단백질들과 함께 β-hexosaminidase를 분비하는 것으로 알려져 있으며 이는 현재 알레르기 측정 지표로 사용되고 있다[Korea J. Herbol. 26, 1-7]. 비만세포의 탈과립을 유도하는 인자들 중 Compound 48/80은 formaldehyde에 의한 cross-linked phenethylamine의 혼합 다당체로서, 비만세포의 막수용체에 작용하여 세포 외의 칼슘을 세포 내로 유입시킴으로써 세포 내 자유칼슘의 양을 증가시켜 비만세포를 탈과립 시키고 히스타민과 세로토닌, 가수분해 효소 등의 매개물질들이 비만세포 밖으로 분비된다[Korean J. Immunol. 21, 35-45]. 따라서 본 발명에서는 장수풍뎅이 유충의 추출물의 항알레르기 효능을 알아보기 위해 RBL-2H3 세포주에 Compound 48/80로 자극 후 이들 세포로부터 유리되는 β-hexosaminidase의 유리양을 측정함으로써 장수풍뎅이 유충의 추출물이 알레르기 반응에 미치는 현상인 비만세포의 탈과립을 확인하였다. When cells are stimulated, various inflammatory substances are secreted. Among them, mast cells are known to secrete β-hexosaminidase along with various proteins present in intracellular granules, which is now used as an indicator of allergy [ J. Herbol. 26, 1-7]. Among the factors inducing degranulation of mast cells, Compound 48/80 is a mixed polysaccharide of cross-linked phenethylamine due to formaldehyde. It acts on the membrane receptor of mast cells and injects extracellular calcium into cells, And degranulation of mast cells, and mediators such as histamine, serotonin, and hydrolytic enzymes are secreted out of mast cells [Korean J. Immunol. 21,35-45]. Therefore, in order to investigate the antiallergic effect of the extracts of long-bred beetle larvae, the present invention is to measure the release of β-hexosaminidase released from these cells after stimulation with Compound 48/80 in RBL-2H3 cell line, And the degranulation of mast cells was observed.
이를 위해 RBL-2H3 세포를 PBS(phosphate buffered saline)에 세척 및 현탁하고 1x106cells/㎖의 농도에서 200 ㎕씩 분주한 후 장수풍뎅이 유충의 추출물을 처리하여 5% CO2조건하에서 37℃로 유지되는 배양기에서 30분 배양하였다. 이후 알레르기 유발 물질인 Compound 48/80을 25 ㎍/㎖ 처리하여 5% CO2 조건하에서 37℃로 유지되는 배양기에서 30분 배양하여 알레르기를 유발하였다. 이 중 100 ㎕의 상등액을 취한 후 β-hexosaminidase ELISA kit (Cloud-Clone Corp., USA)을 이용하여 β-hexosaminidase의 유리량을 측정하였으며 최종 확인은 multi detector (Beckman, DTX 8800, CA, USA)를 이용하여 450 nm 파장에서 흡광도를 확인하였고 이에 대한 결과를 도 1에 나타내었다. For this washing and suspending the RBL-2H3 cells in the (phosphate buffered saline) PBS and kept at 37 ℃ under 1x10 6 cells / ㎖ 5% CO 2 condition and then dispensed at a concentration by 200 ㎕ by treating the extract of the beetle larvae Lt; / RTI > for 30 min. Subsequently, Compound 48/80, which is an allergen, was treated with 25 μg / ml and incubated in an incubator maintained at 37 ° C under 5% CO 2 for 30 minutes to induce allergy. Hexosaminidase was measured by β-hexosaminidase ELISA kit (Cloud-Clone Corp., USA) after taking 100 μl of the supernatant. The final detection was done using a multi detector (Beckman, DTX 8800, CA, USA) The absorbance was confirmed at a wavelength of 450 nm and the results are shown in FIG.
도 1을 참고하면, 장수풍뎅이 유충의 추출물을 125, 250, 500 ㎍/㎖의 농도로 처리한 결과 125 ㎍/㎖에서는 대조군과 비교하여 유의적 차이를 보이지 않았으나 250 ㎍/㎖ 과 500 ㎍/㎖ 농도를 처리하였을 때 β-hexosaminidase 유리율이 현저하게 감소되었다. 특히 500 ㎍/㎖ 농도 조건에서는 무처리 대조군과 거의 유사한 조건으로 β-hexosaminidase 분비가 억제되었다. 이 실험 결과로 장수풍뎅이 유충의 추출물은 RBL-2H3 세포에 농도의존적으로 β-hexosaminidase의 분비를 억제함으로써 항알레르기 효능을 나타내는 것으로 판단된다. As shown in FIG. 1, when the extracts of L. longid bug larvae were treated at a concentration of 125, 250 and 500 μg / ml, there was no significant difference at 125 μg / ml compared to the control group, but 250 μg / ml and 500 μg / The concentration of β-hexosaminidase was significantly decreased when the concentration was treated. Especially, at the concentration of 500 ㎍ / ㎖, the secretion of β-hexosaminidase was suppressed under similar condition to the untreated control group. As a result of this experiment, it was judged that the extract of L. caulifera larvae showed antiallergic effect by inhibiting the secretion of β-hexosaminidase in a concentration-dependent manner on RBL-2H3 cells.
<실시예 4. Cytokine 측정> ≪ Example 4: Cytokine assay >
비만세포가 활성화 되면 비만세포는 탈과립을 통하여 아라키돈산 대사물질과 염증반응을 유발하는 다양한 cytokine이 분비되는데, 그 중 IL-4는 B-cell을 활성화하여 Immunoglobulin E (IgE) 항체를 생성하며, Th2 세포의 분화를 촉진시키므로 천식과 같은 알레르기 질환에서 중요한 역할을 하는 것으로 보고되어 있다[J. Allergy Clin. Immunol. 99, 155-160]. TNF-α는 비만세포와 대식세포 등에서 생성되어 많은 염증반응을 유발하는 인자로, 알레르기 염증반응에 중요한 역할을 하는 것으로 알려져 있는 cytokine 이다[Immunol Today 11, 458-464]. 또한 면역세포의 대표적인 알레르기 염증인자로 알려진 COX (cyclooxygenase)는 COX-1과 COX-2가 두 가지로 존재하는데 COX-1은 대부분의 조직에 존재하며, 신장의 혈액흐름을 조절하거나 위장의 세포를 보호하는 등의 생리적인 기능을 조절한다. 반면 COX-2는 미생물에 의한 감염이나 손상 혹은 여러 요인의 스트레스에 반응하여 발현되어 prostaglandins (PG)의 합성에 관여하고, LPS 및 cytokine에 의해 염증 관련 질병을 유발하는 것으로 밝혀져 있다[Trends Immunol. 23, 144-150; Ann. N. Y. Acad. Sci. 714, 136-42].When mast cells are activated, mast cells secrete various cytokines that induce inflammation reaction with arachidonic acid metabolism through degranulation. Among them, IL-4 activates B-cell to produce immunoglobulin E (IgE) Has been reported to play an important role in allergic diseases such as asthma as it promotes differentiation of cells [ J. Allergy Clin. Immunol. 99, 155-160]. TNF-α is a cytokine that is known to play an important role in the allergic inflammatory response, which is produced by mast cells and macrophages and causes many inflammatory responses [ Immunol Today 11, 458-464]. COX-1 and COX-2 exist in most tissues, and they control the blood flow of the kidneys, and the cells of the gastrointestinal tract And controlling the physiological functions such as protection. On the other hand, COX-2 has been shown to be expressed in response to microbial infection, damage or stress of various factors, involved in the synthesis of prostaglandins (PG), and induced inflammation-related diseases by LPS and cytokines [ Trends Immunol. 23, 144-150; Ann. NY Acad. Sci. 714, 136-42].
본 발명의 장수풍뎅이 유충 추출물이 호흡기질환과 관련된 알레르기 억제 효과가 있는지 확인하기 위해, 먼저 호흡기질환 천식 조건을 비만 세포주 RBL-2H3에 유도하였다. 이를 위해 RBL-2H3 세포를 PBS에 현탁하고 1x106 cells/㎖의 농도에서 200 ㎕씩 1.5 ㎖ tube에 분주한 후 장수풍뎅이 유충의 추출물을 125, 250, 500 ㎍/㎖의 농도별로 처리하여 5% CO2조건하에서 37℃로 유지되는 배양기에서 30분 배양하였다. 이후 Compound 48/80을 25 ㎍/㎖ 처리하여 5% CO2 조건하에서 37℃로 유지되는 배양기에서 30분 배양하였다. 배양 후 상등액을 이용하여 IL-4 (Rat IL-4 ELISA Kit, novex, MA, USA), TNF-α (Rat TNF-α ELISA Kit, Invitrogen, MA, USA), COX-2 (Rat Cyclooxygenase-2 (COX-2) ELISA Kit, CUSABIO, MA, USA) 분비량을 multi detector (Beckman, DTX 8800, CA, USA)를 이용하여 450 nm 파장에서 흡광도를 측정하였다.In order to determine whether the extract of the long-bred beetle larvae of the present invention has an effect of inhibiting allergic diseases associated with respiratory diseases, respiratory disease asthmatic conditions were induced in an obesity cell line RBL-2H3. For this purpose, RBL-2H3 cells were suspended in PBS, and 200 μl of the suspension was dispensed into a 1.5 ml tube at a concentration of 1 × 10 6 cells / ml. The extracts of the larva bug larvae were treated with 125, 250 and 500 μg / Lt; RTI ID = 0.0 > 37 C < / RTI > Compound 48/80 was then treated with 25 μg / ml and incubated for 30 min in an incubator maintained at 37 ° C under 5% CO 2 . (Rat IL-4 ELISA Kit, novex, MA, USA), TNF-α (Rat TNF-α ELISA Kit, Invitrogen, MA, USA) and COX-2 (Rat Cyclooxygenase-2 (COX-2) ELISA Kit, CUSABIO, MA, USA) was used to measure the absorbance at 450 nm using a multi detector (Beckman, DTX 8800, CA, USA).
도 2 및 도 3을 참고하면, Compound 48/80만 처리한 샘플에 비교하여 장수풍뎅이 유충의 추출물을 농도별로 처리하였을 때 농도 의존적으로 IL-4, TNF-α, COX-2 분비량이 모두 감소되는 것을 확인할 수 있으며 특히 모든 조건에서 125 ㎍/㎖의 농도에서도 무처리 대조군과 유사할 정도로 사이토카인이 현저하게 억제됨을 알 수 있다. 따라서 각 cytokine의 분비가 장수풍뎅이 유충의 추출물에 의해 유의적으로 감소된 것을 통해 비만세포가 Th2 세포의 기능을 획득하는 것을 억제함으로써 항천식, 항알레르기 작용을 수행하는 것으로 판단된다. 2 and 3, when IL-4, TNF-α, and COX-2 secretion levels were all reduced in a concentration-dependent manner when the extract of Longevity Beetle larvae was treated by concentration compared to the sample treated with Compound 48/80 alone And especially at a concentration of 125 [mu] g / ml under all conditions, the cytokine is remarkably inhibited to a degree similar to that of the untreated control group. Therefore, the secretion of each cytokine is significantly reduced by the extract of the larvae of beetle larvae, and it is judged that the mast cell performs anti - asthma and anti - allergic action by inhibiting the acquisition of the function of Th2 cell.
Claims (8)
상기 추출물은 장수풍뎅이 유충의 열수 추출물인 것을 특징으로 하는 항알레르기성 조성물.The method according to claim 1,
Wherein the extract is a hot-water extract of a long-lived beetle larva.
상기 추출물은 β-헥소사미노다아제(β-hexosaminidase), IL-4(interleukin-4), COX-2(Cyclooxygenase-2) 또는 TNF-α(Tumor necrosis factor-α)의 분비를 억제하는 효능이 있는 것을 특징으로 하는 항알레르기성 조성물.The method according to claim 1,
The above extract has an efficacy of suppressing the secretion of? -Hexosaminidase, IL-4 (interleukin-4), COX-2 (cyclooxygenase-2) or TNF-α (Tumor necrosis factor- Lt; RTI ID = 0.0 > anti-allergic < / RTI >
상기 질환은 아나필락시스 쇼크, 식품 알레르기, 알레르기성 비염, 화분증, 천식, 약제 알레르기, 식물 알레르기, 두드러기, 습진 및 알레르기성 접촉성 피부염에서 선택되는 알레르기 질환인 것을 특징으로 하는 알레르기 질환의 예방 또는 치료용 약학 조성물.5. The method of claim 4,
The disease is an allergic disease selected from anaphylactic shock, food allergy, allergic rhinitis, hay fever, asthma, drug allergy, plant allergies, urticaria, eczema and allergic contact dermatitis. Composition.
상기 질환은 아나필락시스 쇼크, 식품 알레르기, 알레르기성 비염, 화분증, 천식, 약제 알레르기, 식물 알레르기, 두드러기, 습진 및 알레르기성 접촉성 피부염에서 선택되는 알레르기 질환인 것을 특징으로 하는 예방 또는 개선용 건강기능식품.The method according to claim 6,
Wherein said disease is an allergic disease selected from anaphylactic shock, food allergy, allergic rhinitis, hay fever, asthma, drug allergy, plant allergies, urticaria, eczema and allergic contact dermatitis.
상기 건강기능식품은 각종 드링크제, 육류, 소세지, 빵, 캔디류, 스넥류, 면류, 아이스크림, 유제품, 스프, 이온음료, 음료수, 알코올 음료, 껌, 차 및 비타민 복합제로 이루어진 군에서 선택되는 것을 특징으로 하는 예방 또는 개선용 건강기능식품.The method according to claim 6,
The health functional food is selected from the group consisting of various drinks, meats, sausages, bread, candy, snacks, noodles, ice cream, dairy products, soups, ionic drinks, beverages, alcoholic beverages, gums, tea and vitamins Health functional foods for prevention or improvement.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR102158752B1 (en) * | 2019-06-27 | 2020-09-22 | 세종대학교산학협력단 | A pharmaceutical composition comprising a Allomyrina dichotoma extract and a health functional food |
| EP4140319A4 (en) * | 2020-04-23 | 2024-05-22 | CJ Cheiljedang Corporation | Composition comprising beetle larva or extract thereof as active ingredient for improving bowel movement function |
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| KR102024188B1 (en) | 2019-09-23 |
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