KR20190021614A - Composition for antioxidant, antiinflammatory and inflammatory neurodegenerative diseases comprising perilla frutescens britton extract - Google Patents
Composition for antioxidant, antiinflammatory and inflammatory neurodegenerative diseases comprising perilla frutescens britton extract Download PDFInfo
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- KR20190021614A KR20190021614A KR1020170106558A KR20170106558A KR20190021614A KR 20190021614 A KR20190021614 A KR 20190021614A KR 1020170106558 A KR1020170106558 A KR 1020170106558A KR 20170106558 A KR20170106558 A KR 20170106558A KR 20190021614 A KR20190021614 A KR 20190021614A
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- perilla
- red ginseng
- inflammatory
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- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/53—Lamiaceae or Labiatae (Mint family), e.g. thyme, rosemary or lavender
- A61K36/535—Perilla (beefsteak plant)
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- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/105—Plant extracts, their artificial duplicates or their derivatives
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- A—HUMAN NECESSITIES
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Abstract
Description
본 발명은 생들깨박 추출물을 유효성분으로 포함하는 항산화, 항염증 및 염증성 신경퇴행성 질환 예방 또는 치료용 조성물에 관한 것이다. The present invention relates to a composition for preventing or treating antioxidant, antiinflammatory and inflammatory neurodegenerative diseases, which comprises a raw perilla oil extract as an active ingredient.
들깨(Perilla frutescens)는 꿀풀과(Laviatae)에 속하는 일 년생 초본으로 들깨종자는 들기름, 차, 죽, 제과 등에 사용되며 엽채는 신선채고, 절임, 통조림 등에 사용되고 있다(Choung, M. G., Comparison of major characteristics between seed perilla and vegetable perilla, Korean J. Crop. Sci., 50, 171-174, 2005).Perilla frutescens is a perennial plant belonging to the genus Laviatae. Perilla seeds are used for perilla, tea, porridge, and confectionery. Leaves are used for fresh picking, pickling, canning, etc. (Choung, MG, Comparison of major characteristics between seed perilla and vegetable perilla, Korean J. Crop. Sci., 50, 171-174, 2005).
또한, 들깨는 일반 성숙 채소보다 종자가 발아되면서 아미노산, 지방산, 탄수화물 등 영양성분의 변화와 함께 비타민, 무기질 그리고 폴리페놀 등 항산화 효과를 나타내는 물질의 함량이 종자 및 성숙기에 비해 높아 생리활성물질이 증대되는 것으로 알려져 건강 기능성 식품으로 인정받고 있다(Khalil, A.W. et al., Comparison of sprout quality characteristics of desi and kabuli type chickpea cultivars (Cicer arietinum L.), LWTFood Sci. Technol., 40, 937-945, 2007).In addition, perilla seeds germinated more than ordinary mature vegetables, and the contents of amino acids, fatty acids, carbohydrates, and other substances showing antioxidative effects such as vitamins, minerals and polyphenols are higher than those of seeds and maturity, (Cicer arietinum L.), LWTFood Sci. Technol., 40, 937-945, 2007, which is recognized as a health functional food (Khalil, AW et al., Comparison of sprout quality characteristics of desi and acceptance type chickpea cultivars ).
뿐만 아니라, 들깨는 오메가-3 지방산인 리놀렌산이라든지 비타민이 풍부하게 함유되어 있어 인체의 두뇌활동을 촉진시켜 치매의 예방에 도움을 주고 조혈작용이 우수하여 빈혈과 저혈압의 예방에 효과적일 뿐만 아니라 혈중콜레스테롤을 저감시켜 동맥경화의 예방이나 항염증에 유리한 효능을 나타내고 피부 미용과 노화방지에도 탁월환 효능을 갖는다.In addition, the perilla is rich in omega-3 fatty acids, linolenic acid or vitamins, which promotes the brain activity of the human body and helps to prevent dementia and hematopoiesis because it is effective in preventing anemia and hypotension, as well as blood cholesterol Which is effective for prevention of arteriosclerosis and anti-inflammation, and has an excellent circulating effect for preventing skin beauty and aging.
상기한 생리활성물질과 효능을 가지는 들깨는 전통적인 약용작물(medicinal herb) 중의 하나로 수세기 동안 우울증, 종양, 기침, 항균, 항곰팡이, 알러지 등 다양한 질병을 다스리는데 사용되어 왔다. The above-mentioned physiologically active substance and perilla having efficacy have been used as medicinal herb for centuries to control various diseases such as depression, tumor, cough, antibacterial, antifungal and allergy.
즉, 들깨의 정유성분(essential oil)은 항곰팡이 활성이 높은 것으로 보고되어으며, 들깨의 주요 생리활성물질은 tormentic acid (TA), oleanolic acid (OA) 그리고 ursolic acid (UA)로 알려져 있다. 이 중 TA는 항염증 효과와 DNA polymerase 저해활성이 있는 것으로 보고되어 있고, OA와 UA 또한 항염증 효과가 있는 것으로 보고되어 있으며, OA는 간 보호 효과와 항종양효과가 있는 것으로 보고되고 있다.The major physiologically active substances of perilla are known as tormentic acid (TA), oleanolic acid (OA) and ursolic acid (UA). TA has been reported to have antiinflammatory and DNA polymerase inhibitory activity, and OA and UA have also been reported to have anti-inflammatory effects. OA has been reported to have hepatoprotective and antitumor effects.
한편, Superoxide dismutase (SOD), glutathione peroxidase (GPX), catalase (CAT), glutathione reductase, glutathione-S-transferase 등과 같이 인체 내에는 활성산소에 방어하는 항산화 효소를 가지고 있지만 산업화와 더불어 증가되는 각종 환경오염 물질, 흡연, 스트레스 등에 의해 인체 내의 항산화계의 역할만으로는 방어 체계가 초과되어 단백질 분해, DNA 손상 등이 유발된다 On the other hand, there are antioxidant enzymes that protect against reactive oxygen species in the body such as superoxide dismutase (SOD), glutathione peroxidase (GPX), catalase (CAT), glutathione reductase and glutathione-S- transferase. However, The role of antioxidants in the human body due to substances, smoking, stress, etc., exceeds the defense system, leading to protein degradation and DNA damage
따라서, 항산화제로 butylated hydroxyanisole (BHA)과 butylated hydroxytoluene(BHT)와 같은 합성물질들이 개발되었으나, 이들의 경우 다량을 섭취하면 여러 가지 부작용을 나타낼 수 있어 천연으로부터 안전한 식이성 항산화물질을 찾는 연구가 활발히 진행되고 있다Therefore, synthetic materials such as butylated hydroxyanisole (BHA) and butylated hydroxytoluene (BHT) have been developed as antioxidants. However, studies on finding dietary antioxidants that are safe from nature have been actively carried out Become
또 다른 한편, 염증은 감염으로 인한 인체 조직손상을 막는 방어기전으로 발전, 발열, 통증과 같은 증상을 수반한다. 이러한 염증반응이 장기간 지속적으로 반복될 경우 신경퇴행성질환과 같은 각종 만성질환과 암 등을 발병할 수 있는 요인이 된다. On the other hand, inflammation is accompanied by symptoms such as fever, fever, and pain before the defense period to prevent human tissue damage from infection. If the inflammatory reaction is repeated for a long period of time, it can cause various chronic diseases such as neurodegenerative diseases and cancer.
특히, 인체 내 대식세포(macrophage)는 인체 내 면역반응에서 일산화질소(nitric oxide; NO)와 프로스타글란딘(prostaglandin; PG)과 pro-inflammatory cytokine 등과 같은 염증매개물질 생성에 관여하고 이를 조절한다. 이러한 염증매개물질은 염증반응을 유도하며, 숙주의 면역반응이 적절하게 대응하지 못할 경우 염증성 질환을 유발한다.In particular, macrophages in the human body are involved in and regulate the production of inflammatory mediators such as nitric oxide (NO), prostaglandin (PG) and pro-inflammatory cytokines in the human immune response. These inflammatory mediators induce an inflammatory response and, if the host immune response does not respond appropriately, induce inflammatory diseases.
상기한 들깨의 항산화 및 항염증효과를 이용한 종래 기술들을 살펴 보면, 한국등록특허 10-0614970(2006년08월16일)에 체내 지질과산화물의 축적을 억제하고, 동시에 항산화효소의 활성을 증가시키는 효과가 있으며, 또한 신경전달체계에 있어서 매우 중요한 역할을 수행하는 아세틸콜린에스터라아제의 활성을 저하하는 방법으로 치매 등의 인지기능장애를 예방하고, 동시에 개선시키는 효과가 우수하여 다양한 건강보조식품 내지는 약제학적 조성물로서 제공될 수 있는 들깨박 추출물을 유효성분으로 함유하는 인지기능장애예방 및 개선용 조성물이 공지되어 있다.The above-mentioned prior arts using the antioxidative and anti-inflammatory effects of perilla are described in Korean Patent No. 10-0614970 (Aug. 16, 2006), which suppresses the accumulation of lipid peroxides in the body and simultaneously increases the activity of antioxidant enzymes In addition, the method of decreasing the activity of acetylcholinesterase, which plays a very important role in the neurotransmission system, is effective in preventing and curing cognitive dysfunction such as dementia, There is known a composition for preventing and improving cognitive dysfunction, which contains perilla oil extract as an active ingredient which can be provided as a pharmaceutical composition.
또한, 한국등록특허 10-1153870(2012년05월31일)에 뉴라미니데이즈 억제 활성을 가지는 들깨과(科) 식물의 추출물 및 이를 유효성분으로 포함하는 인플루엔자 예방 또는 치료용 조성물로서, 들깨 추출물에 포함된 폴리페놀계 화합물들이 뉴라미니데이즈를 억제하여 인플루엔자를 예방, 치료할 수 있는 조성물, 건강식품, 가축 사료 첨가제 등을 제공할 수 있도록 한 뉴라미니데이즈 억제활성을 가지는 들깨 추출물이 공지되어 있다.Also, Korean Patent No. 10-1153870 (May 31, 2012) discloses an extract of a perilla plant having an inhibitory activity against neuraminidaysis, and a composition for preventing or treating influenza comprising the same as an active ingredient. A perilla extract having a neuraminidase inhibitory activity, which is capable of providing a composition capable of preventing and treating influenza by inhibiting neuraminidase, a health food, a livestock feed additive and the like, are known.
또한, 한국등록특허 10-1440372(2014년09월04일)에는 들깨새싹 추출물을 염증성 질환의 예방 및 치료를 위한 약학조성물로 사용할 수 있을 뿐만 아니라 식품류 및 차 등의 기능성 식품으로 유용하게 사용할 수 있는 들깨 새싹 추출물을 유효성분으로 함유하는 항염증용 식품 및 약학 조성물이 공지되어 있다.Korean Patent No. 10-1440372 (Sep. 04, 2014) discloses that perilla seed extract can be used not only as a pharmaceutical composition for the prophylaxis and treatment of inflammatory diseases but also as a functional food such as foods and tea An anti-inflammatory food and pharmaceutical composition containing perilla sprout extract as an active ingredient is known.
또한, 한국공개특허 10-2017-0024864(2017년03월08일)에는 들깨로부터 박피된 들깨껍질을 원료로서 채취하여 그 들깨껍질원료 내에 포함된 이물질과 오염물을 제거하고 세척수에 의해 세척하며, 그 세척된 들깨껍질원료를 40℃에서 건조시켜 그 건조된 들깨껍질원료를 80∼140℃의 온도조건 하에 대략 10∼15분간 증자시키고, 그 증자된 들깨껍질원료를 온압법으로 착유하고나서 배치(batch)법에 의해 매 압착 시마다 들깨껍질원료를 공급하여 들깨유를 추출하며, 그 추출된 들깨유를 캡슐 등의 적정한 제형방법으로 포장하여 공급함으로써 고품질의 오메가-3 지방산인 리놀렌산을 함유한 들깨유가 다양한 건강기능식품 또는 기능성 제품에 적용이 가능할 수 있도록 한 들깨껍질을 이용한 오메가-3 지방산이 함유된 추출물 제조방법이 공지되어 있다.In Korean Patent Laid-Open Publication No. 10-2017-0024864 (Mar. 08, 2017), peeled perilla peel from perilla is taken as a raw material, and foreign substances and contaminants contained in the perilla peel raw material are removed, washed with washing water, The washed perilla skin raw material is dried at 40 DEG C, the dried perilla skin raw material is heated to a temperature of 80 to 140 DEG C for about 10 to 15 minutes, and the thus-grown perilla skin raw material is milked by pressure method, ) Method, the perilla oil is extracted by supplying the perilla skin raw material at each pressing, and the extracted perilla oil is packed and packed in a proper formulation method such as capsules, so that the perilla oil containing the high quality omega-3 fatty acid, linolenic acid, A method for producing omega-3 fatty acid-containing extracts using perilla peels that can be applied to health functional foods or functional products is known.
또한, 한국등록특허 10-1694660(2017년01월03일)에는 들깨종자를 암(暗)의 조건하에서 12시간, 16 ~ 20℃로 2일간 최아 한 후 8일간 재배하여 들깨새싹을 얻는 단계; 상기 단계에서 얻은 들깨새싹을 음건하여 프로판디올을 첨가하여 35 ~ 40℃에서 초음파추출하는 단계로 이루어진 항염증 효과가 있는 들깨새싹 추출물의 제조방법이 공지되어 있다.Korean Patent No. 10-1694660 (Jan. 03, 2017) discloses a method for growing perilla seedlings, which comprises growing perilla seedlings under dark conditions for 12 hours, at 16-20 ° C for 2 days and then for 8 days to obtain perilla sprouts; The perilla sprouts obtained in the above step are shaded, and propanediol is added thereto and ultrasonically extracted at 35 to 40 ° C. There is known a method for producing the perilla sprout extract having an anti-inflammatory effect.
그러나, 상기한 들깨 추출물들은 속성상 항산화 또는 항염증효과를 가지기는 하나, 아세틸콜린에스터라아제의 활성을 저하하는 방법 인지기능을 보조한다거나, 인플루엔자 예방 또는 치료, 아토피 등의 피부염증 완화를 위한 것이고, 오메가-3 지방산이 함유된 추출물 또는 들깨새싹 추출물의 제조방법에 관한 것일 뿐, 항산화, 항염증 또는 염증성 신경퇴행성 질환 예방 또는 치료용 조성물로 사용하는 것은 불가능한 문제점이 있었다.However, the perilla extracts described above have an antioxidant or antiinflammatory effect on the nature, but they are intended to assist the method for reducing the activity of acetylcholinesterase, to prevent or treat influenza, to alleviate skin inflammation such as atopy, The present invention relates to a method for producing omega-3 fatty acid-containing extracts or perilla sprout extracts, and has a problem in that it can not be used as a composition for preventing or treating antioxidant, anti-inflammatory or inflammatory neurodegenerative diseases.
본 발명은 상기의 문제점을 해결하기 위하여, 생들깨박 추출물을 유효성분으로 포함하는 항산화 조성물을 제공하는 것을 해결하고자 하는 과제로 한다.DISCLOSURE OF THE INVENTION An object of the present invention is to provide an antioxidative composition comprising a perilla perilla extract as an active ingredient in order to solve the above problems.
또한, 본 발명은 상기의 문제점을 해결하기 위하여, 생들깨박 추출물을 유효성분으로 포함하는 항염증 조성물을 제공하는 것을 해결하고자 하는 과제로 한다.Another object of the present invention is to provide an anti-inflammatory composition comprising a perilla perilla extract as an active ingredient to solve the above problems.
또한, 본 발명은 생들깨박 추출물을 유효성분으로 포함하는 염증성 신경퇴행성 질환 예방 또는 치료용 조성물을 제공하는 것을 해결하고자 하는 과제로 한다.The present invention also provides a composition for the prevention or treatment of inflammatory neurodegenerative diseases, which comprises a perilla plant extract as an active ingredient.
본 발명은 상기 과제를 해결하기 위하여, 생들깨박 추출물을 유효성분으로 포함하는 항산화 조성물을 과제의 해결수단으로 한다.In order to solve the above problems, the present invention provides an antioxidant composition comprising a perilla perilla extract as an active ingredient.
또한, 본 발명은 생들깨박 추출물을 유효성분으로 포함하는 항염증 조성물을 과제의 해결수단으로 한다.The present invention also provides an anti-inflammatory composition comprising a perilla perilla extract as an active ingredient as a means for solving the problem.
또한, 본 발명은 생들깨박 추출물을 유효성분으로 포함하는 염증성 신경 퇴행성 질환 예방 또는 치료용 조성물을 과제의 해결수단으로 한다.The present invention also provides a composition for the prevention or treatment of inflammatory neurodegenerative diseases, which comprises a perilla plant extract as an active ingredient.
상기 조성물은 홍삼박 추출물을 더 포함하는 것을 과제의 해결수단으로 한다.The composition of the present invention further comprises a red ginseng extract.
상기 생들깨박 추출물 및 홍삼박 추출물은 생들깨박 에탄올 추출물 및 홍삼박 에탄올 추출물인 것을 과제의 해결수단으로 한다.The raw perilla peruvianum extract and the red ginseng root extract are the perennial perilla ethanol extract and the ethanol extract of red ginseng.
상기 조성물은 유도성 질소 산화물 합성(iNOS) 및 NO 생산을 약화시키는 것을 과제의 해결수단으로 한다.The composition makes the solution to the task of attenuating inductive nitrogen oxide synthesis (iNOS) and NO production.
상기 조성물은 미세아교세포에서 미세아교세포 활성을 억제하는 것을 과제의 해결수단으로 한다.The above composition provides a solution to the problem of inhibiting microglial cell activity in microglia.
상기 조성물은 항산화 및 항염증 활성 보조성분으로 석류추출액 및 프로폴리스를 더 포함하는 것을 과제의 해결수단으로 한다.The composition of the present invention further comprises a pomegranate extract and propolis as an antioxidant and anti-inflammatory activity auxiliary ingredient.
또한, 본 발명은 상기 생들깨박 추출물을 유효성분으로 포함하는 항산화 조성물, 항염증 조성물 또는 염증성 신경 퇴행성 질환 예방 또는 치료용 조성물을 포함하는 식품 조성물을 과제의 해결수단으로 한다.The present invention also provides a food composition comprising an antioxidative composition, an anti-inflammatory composition or a composition for the prevention or treatment of inflammatory neurodegenerative diseases, which comprises the perilla plant extract as an active ingredient.
상기 식품 조성물은 음료, 육류, 초코렛, 과자류, 피자, 라면, 면류, 껌류, 캔디, 젤리, 아이스크림류, 유제품, 알코올 음료류, 비타민 복합제인 것을 과제의 해결수단으로 한다.The food composition is a solution to the problem of being a beverage, a meat, a chocolate, a confectionery, a pizza, a ramen, a noodle, a gum, a candy, a jelly, an ice cream, a dairy product, an alcoholic beverage, and a vitamin complex.
본 발명에 따른 생들깨박 추출물을 유효성분으로 포함하는 항산화, 항염증 및 염증성 신경퇴행성 질환 예방 또는 치료용 조성물은 BV-2 세포에 있어서 LPS-유발 유도성 질소 산화물 합성(iNOS) 및 NO 생산을 약화시키고, 미세아교세포에서 미세아교세포 활성을 억제함으로써, 효율적인 항산화제, 항염증제 및 염증성 신경퇴행성 질환의 예방 및 치료제로 사용될 수 있는 우수한 효과가 있다.The composition for preventing or treating antioxidant, anti-inflammatory and inflammatory neurodegenerative diseases comprising the perilla perilla extract according to the present invention as an active ingredient is useful for the production of LPS-inducible nitrogen oxides (iNOS) and NO production in BV-2 cells And inhibiting microglial cell activity in microglial cells. Thus, there is an excellent effect that can be used as an effective antioxidant, an anti-inflammatory agent and an agent for the prevention and treatment of inflammatory neurodegenerative diseases.
도 1은 생들깨박 추출물의 DPPH 저해능을 나타낸 그림.
도 2는 생들깨박 추출물의 세포생존율(BV-2 세포)을 나타낸 그림.
도 3은 생들깨박 추출물의 NO 생성 저해를 나타낸 그림.
도 4는 들깨박, 홍삼박 복합추출물의 DPPH 라디칼 저해효능을 나타낸 그림FIG. 1 shows the DPPH inhibitory effect of fresh perilla perilla extract.
Figure 2 shows the cell viability (BV-2 cells) of fresh perilla perilla extract.
FIG. 3 is a graph showing inhibition of NO production by fresh perilla plant extract.
Fig. 4 shows the DPPH radical inhibitory effect of the perilla and red ginseng compound extracts
본 발명은, 생들깨박 추출물을 유효성분으로 포함하는 항산화 조성물을 기술구성의 특징으로 한다.The present invention is characterized by the technical composition of an antioxidant composition comprising a perilla perilla extract as an active ingredient.
또한, 본 발명은 생들깨박 추출물을 유효성분으로 포함하는 항염증 조성물을 기술구성의 특징으로 한다.In addition, the present invention is characterized by the technical composition of an anti-inflammatory composition comprising a perilla perilla extract as an active ingredient.
또한, 본 발명은 생들깨박 추출물을 유효성분으로 포함하는 염증성 신경 퇴행성 질환 예방 또는 치료용 조성물을 기술구성의 특징으로 한다.The present invention also provides a composition for preventing or treating an inflammatory neurodegenerative disease, comprising a perilla perilla extract as an active ingredient.
상기 조성물은 홍삼박 추출물을 더 포함하는 것을 기술구성의 특징으로 한다.The composition of the present invention further comprises a red ginseng extract.
상기 생들깨박 추출물 및 홍삼박 추출물은 생들깨박 에탄올 추출물 및 홍삼박 에탄올 추출물인 것을 기술구성의 특징으로 한다.The raw perilla perilla extract and the red ginseng root extract are characterized in that the perilla perilla ethanol extract and the ethanol extract of red ginseng are ethanol extracts.
상기 조성물은 유도성 질소 산화물 합성(iNOS) 및 NO 생산을 약화시키는 것을 기술구성의 특징으로 한다.The composition is characterized by the technical constitution that it weakens inductive nitrogen oxide synthesis (iNOS) and NO production.
상기 조성물은 미세아교세포에서 미세아교세포 활성을 억제하는 것을 기술구성의 특징으로 한다.The composition is characterized in that it inhibits microglial cell activity in microglial cells.
상기 조성물은 항산화 및 항염증 활성 보조성분으로 석류추출액 및 프로폴리스를 더 포함하는 것을 기술구성의 특징으로 한다.Wherein the composition further comprises a pomegranate extract and propolis as an antioxidant and anti-inflammatory activity auxiliary component.
또한, 본 발명은 상기 생들깨박 추출물을 유효성분으로 포함하는 항산화 조성물, 항염증 조성물 또는 염증성 신경 퇴행성 질환 예방 또는 치료용 조성물을 포함하는 식품 조성물을 기술구성의 특징으로 한다.The present invention also features a food composition comprising an antioxidative composition, an anti-inflammatory composition or a composition for the prevention or treatment of inflammatory neurodegenerative diseases, comprising the pericarpella extract as an active ingredient.
상기 식품 조성물은 음료, 육류, 초코렛, 과자류, 피자, 라면, 면류, 껌류, 캔디, 젤리, 아이스크림류, 유제품, 알코올 음료류, 비타민 복합제인 것을 기술구성의 특징으로 한다.Wherein the food composition is characterized by comprising beverage, meat, chocolate, confectionery, pizza, ramen, noodles, gum, candy, jelly, ice cream, milk product, alcoholic beverage, and vitamin complex.
이하에서는 본 발명이 속하는 기술분야에서 통상의 지식을 가진 자가 용이하게 실시할 수 있도록 본 발명을 상세히 설명한다. 그러나 본 발명은 여러 가지 상이한 형태로 구현될 수 있으며, 여기에서 설명하는 것으로 한정되지 않는다.Hereinafter, the present invention will be described in detail in order to facilitate a person skilled in the art to which the present invention pertains. The present invention may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein.
본 발명의 항산화 조성물, 항염증 조성물 또는 염증성 신경 퇴행성 질환 예방 또는 치료용 조성물은 생들깨박 추출물을 유효성분으로 포함하며, 상기 생들깨박 추출물은 생들깨박 에탄올 추출물이다.The antioxidative composition, the anti-inflammatory composition or the composition for preventing or treating inflammatory neurodegenerative diseases according to the present invention comprises a raw perilla oil extract as an active ingredient and the perilla perilla oil extract is a perilla perilla ethanol extract.
여기서, 상기 생들깨박 추출물을 유효성분은 염증에 관여하는 유도성 질소 산화물 합성효소(iNOS) 및 NO 생산을 약화시켜 함염증효과를 나타낸다.Herein, the active ingredient of the perilla perilla extract exhibits an inflammatory effect by weakening the production of inducible nitric oxide synthase (iNOS) and NO, which are involved in inflammation.
상기 유도성 질소 산화물 합성효소(iNOS ; inducible NO synthase)는 대식세포 등인 면역계 세포에서 시토카인 자극으로 발현이 유도되어 L-아르기닌을 기질로서 일(一)산화질소(NO)를 합성하는 130kDa 효소이다. 이 밖에 2종류인 동질효소(isozyme)처럼, C말단측에는 시토크롬P450 환원효소와 공통적으로 인정되는 NADPH, FAD, FMN결합부위가 존재한다. 구성형NO생성효소인 혈관내피형NO생성효소 혹은 신경형NO생성효소와는 달리 카르모듈린(carmodulin)에 대한 친화성이 크기 때문에 효소활성은 칼슘/카르모듈린비의존성으로 주로 전사수준에서 제어되고 있다.The inducible NO synthase (iNOS) is a 130 kDa enzyme that induces expression by cytokine stimulation in macrophage cells such as macrophages and synthesizes nitric oxide (NO) using L-arginine as a substrate. In addition, there are NADPH, FAD and FMN binding sites common to cytochrome P450 reductase on the C-terminal side, as are two isozymes. Since the affinity for carmodulin is high, unlike the NO production enzyme, which is a constitutive NO production enzyme, or the nerve endogenous NO production enzyme, the enzyme activity is mainly controlled at the transcription level by the calcium / carmodulin dependence .
상기 iNOS의 유전자발현 유도는 염증반응과 밀접하게 관여하고 있으며, INF-γ, INF-α, INF-β 등인 염증성 시토카인의 자극으로 대식세포, 평활근세포, 간세포 등인 세포에서 유전자가 유도되어 NO가 생성된다. 감염 또는 염증반응에 따라 iNOS에서 생산되는 과잉NO는 생리가능보다 오히려 여러 가지 질환에 깊이 관여하고 있다.The induction of the gene expression of iNOS is closely related to the inflammatory reaction. The stimulation of inflammatory cytokines such as INF-γ, INF-α and INF-β induces gene expression in cells such as macrophages, smooth muscle cells and hepatocytes, do. Excess NO produced in iNOS by infection or inflammatory reaction is deeply involved in various diseases rather than physiological.
또한, 본 발명의 생들깨박 추출물을 유효성분으로 포함하는 항산화 조성물, 항염증 조성물 또는 염증성 신경 퇴행성 질환 예방 또는 치료용 조성물은 홍삼박 추출물을 더 포함하며, 상기 홍삼박 추출물은 홍삼박 에탄올 추출물이다.In addition, the antioxidative composition, anti-inflammatory composition or composition for preventing or treating inflammatory neurodegenerative disease, which comprises the raw persimmon extract of the present invention as an active ingredient, further comprises red ginseng roots extract, and the red ginseng roots extract is an ethanol extract of red ginseng .
상기 홍삼박 추출물과 관련하여, 현재 우리나라에서는 인삼을 건강기능식품의 원료로 고시하고 있으며, 인삼과 홍삼제품들을 제조할 경우 추출 및 농축을 통해서 농축액을 제조한 후, 필요한 제품의 유형에 따라 농축액, 농축액 분말 그리고 음료 등으로 사용하고 있다. In relation to the above-mentioned red ginseng roots extract, in Korea, ginseng is reported as a raw material for health functional food. When producing ginseng and red ginseng products, the concentrate is prepared through extraction and concentration, and then, Concentrate powders and beverages.
이때, 홍삼 농축액을 제조하기 위하여 추출용매로 열수 또는 주정을 사용하고 있으며 대부분 홍삼의 원형 그대로 열수 또는 주정에 침지하여 유효성분을 추출해내고 있다. In order to prepare red ginseng concentrate, hot water or alcohol is used as an extraction solvent, and most of the red ginseng is directly extracted as hot water or immersed in alcohol to extract active ingredients.
이때 상기 유효성분을 추출하고 남은 찌꺼기인 홍삼박은 상당한 양의 다당체가 용출되지 않고 함유되어 있으나 산업적으로 동물 사료나 퇴비로 이용되거나 대부분은 폐기되고 있는 실정이다. At this time, the red ginseng leaves remaining after extracting the active ingredient contain a considerable amount of polysaccharide not eluted, but they are industrially used as animal feeds, compost, or mostly discarded.
상기 홍삼박이 닭의 사료로 사용되었을 때 산란률과 면역력이 상승하였다는 보고가 있고, 수입의존도가 높은 면실박을 대체하여 버섯재배를 위한 배지로 이용하였을 때, 느타리 버섯의 수량이 증가하고 품질에는 차이가 없는 것으로 알려져 있다.When the red ginseng roots were used as feed for chickens, it was reported that the egg production rate and immunity were increased. When used as a medium for mushroom cultivation, There is no known difference.
따라서 본 발명에서는 홍삼박의 이용가치를 높이고 홍삼박 추출물에 함유된 유효성분을 이용하기 위하여 들깨박 추출물과 혼합하여 사용하는 것이 바람직하다.Therefore, in the present invention, it is preferable to increase the use value of red ginseng roots and to mix the roots with the perilla flavor extract in order to use the active ingredient contained in the red ginseng roots extract.
한편, 본 발명에서는 석류추출액 및 프로폴리스를 항산화 및 함염 활성 보조성분으로 사용한다.Meanwhile, in the present invention, pomegranate extract and propolis are used as an antioxidant and antiseptic activity auxiliary component.
상기 석류추출액은 석류나무과(Punicaceae)의 열매인 석류(Punicagranatum L.)를 추출하여 얻어진 것이다. 석류의 주요 성분으로는 포도당 및 과당이 다량 함유되어 있으며, 구연산(시트르산)을 함유하여 혈액 정화 및 순환작용을 한다. 또한, 칼슘, 칼륨 등 우리 몸에서 중요한 기능을 수행하는 미네랄 성분들을 풍부하게 가지고 있으며, 그 외에 비타민 B1, 비타민 B2, 비타민 C 등의 비타민을 함유하고 있어서 신체 내에서 각종 기능의 조절 및 질병들을 예방하는 효과가 있다.The pomegranate extract was obtained by extracting pomegranate (Punicagranatum L.), a fruit of the pomegranate (Punicaceae). The major components of pomegranate contain a large amount of glucose and fructose. It contains citric acid (citric acid) to purify and circulate blood. In addition, it has abundant minerals that perform important functions in our bodies such as calcium and potassium. It also contains vitamins such as vitamin B1, vitamin B2 and vitamin C to control various functions in the body and prevent diseases .
특히, 석류는 식물성 에스트로겐인 이소플라본이 함유되어 있어 여성호르몬 결핍에 따른 피부 노화 및 건조 현상을 지연시키는 피부의 대사기능도 기대할 수 있는 효과가 있다.Particularly, pomegranate contains isoflavone, which is a phytoestrogen, and it can be expected that the metabolism function of skin which delays skin aging and drying phenomenon due to female hormone deficiency can be expected.
상기 석류추출액은 석류 나무에서 얻어진 열매와 석류의 종자, 껍질, 잎, 줄기 및 싹 등을 분쇄, 추출하여 얻어진 것으로, 석류분쇄물에 정제수를 혼합하여 가열 추출하여 얻어진다. 즉, 석류분쇄물 : 정제수를 1 : 2~10의 중량비로 혼합하고 50~100℃에서 1 내지 24시간 동안 가열하여 추출하고 얻어진 추출액을 냉각 여과하여 제조한다.The pomegranate extract is obtained by pulverizing and extracting the seeds, husks, leaves, stalks and shoots of the fruit and pomegranate obtained from the pomegranate tree. That is, the pulverized product of pomegranate: purified water is mixed at a weight ratio of 1: 2 to 10, heated at 50 to 100 ° C for 1 to 24 hours for extraction, and the resulting extract is subjected to cooling filtration.
또한, 프로폴리스(propolis)는 꿀벌이 자신의 생존과 번식을 위해 다양한 종류의 수목 및 초본으로부터 뽑아낸 수지(樹脂)와 같은 물질을 벌들이 뒷다리에 붙여 모은 후 자신들의 타액과 혼합하여 타액속의 효소와의 반응에 의해 생성되는 다양한 고분자 중합체들의 복합물에 대한 총칭으로서, 그 주성분은 플라보노이드이다. 플라보노이드는 플라본, 플라보논, 플라보놀, 플라바놀, 플라바논 및 아이소플라본 등과 같은 안소잔신류 및 당과 결합된 그 배당체, 수백 종의 안소사이아닌 및 당과 결합된 그 배당체, 탄닌류로서의 캬테킨 및 류우코잔신을 포함한다. 프로폴리스는 B.C. 300년 훨씬 이전부터 고대 이집트인들에 의하여 방부제로서 미이라의 방부 처리에 효과적으로 사용되었으며, 이러한 방부제로서의 효능은 벌집에 나방이나 작은 들쥐 또는 뱀 등이 들어가 벌들의 공격에 의해 죽은 후, 이들을 벌집 밖으로 운반치 못할 경우 벌들에 의해 코팅 처리된 프로폴리스에 의해 수년간 부패하지 않고 원형이 그대로 보존되고 있는 자연적인 관찰 사실로부터 확인될 수 있다. 프로폴리스의 방부 외의 효과로는 항암성, 화상 치유능, 습진, 아토피성 피부염 및 무좀 등과 같은 진균류에 의한 피부병 치유능, 탈모 억제능, 생리통 완화 내지 제거 효과, 변비, 피부미용 및 주름 제거 등과 미용 효과, 충치, 치육염 및 구강염 등과 같은 세균성 치과적 질환에 대한 치유 및 예방능이 보고되고 있으며, 이를 근거로 우리나라 보건복지부에서도 최근 식품공전 13-25에 건강보조식품으로 규정해 놓고 있다. 또한, 프로폴리스의 주성분인 플라보노이드에 대해서는, 1969년 러시아 카바르키나 박사의 5.6 배에 달하는 면역력 상승 효과, 1973년 러시아 알렉산드로플의 페니실린 보다 더 강한 살균력 실험 입증 발표, 1991년 일본생화학회의 진통작용과 항염 효과, 1994년 일본공업전문대학에서의 제암효능 임상실험결과, 기타 전신 세포의 활성화, 호르몬자극 활동, 비타민 P의 작용에 의한 괴혈병 치료, 고혈압 환자의 혈압강하작용, 생체 호르몬의 균형 유지 등의 효과에 대한 세계 여러 나라의 연구 결과 및 임상시험결과가 보고되어 있다.In addition, propolis is a type of bee that collects materials such as resin (resin) extracted from various kinds of trees and herbs in order to survive and reproduce itself by attaching bees to their hind legs and then mixing them with their saliva, A generic term for a complex of various polymeric polymers produced by the reaction of a polymer with a polymeric material, its principal component being a flavonoid. Flavonoids include, but are not limited to, anosoxenes such as flavone, flavanone, flavonol, flavanol, flavanone and isoflavone and their glycosides conjugated with sugars, hundreds of anthocyanins and their glycosides conjugated with sugars, And Ryuukosan ginseng. Propolis is B.C. It has been used by ancient Egyptians for over 300 years as an antiseptic to be used as an antiseptic for mummified antimicrobial preservatives. The efficacy of this antiseptic is due to the fact that moths, small animals or snakes, It can be confirmed from the natural observations that the original shape is preserved without being corrupted by the propolis coated by bees for many years. The effects other than preservation of propolis include cosmetic effects such as anti-cancer effect, burn healing ability, eczema, atopic dermatitis and fungus-like fungi healing ability, anti-alopecia suppression ability, mitigating or eliminating the dysmenorrhea, constipation, , Tooth decay, dental caries, and oral salt. Based on this, Korea's Ministry of Health and Welfare has recently defined as food supplement 13-25 as food supplement. In addition, for flavonoids, which are the main component of propolis, there was an increase in immunity of 5.6 times that of Dr. Kabarukina of Russia in 1969, a demonstration of stronger sterilization power than that of penicillin in Alexanderplatz in 1973, Anti-inflammatory effect, clinical trial results of ammonia efficacy at Japan Institute of Technology in 1994, activation of other systemic cells, hormone stimulation activity, treatment of scurvy by action of vitamin P, hypotensive effect of hypertensive patients, maintenance of biohormone balance Results of studies and clinical trials in various countries around the world have been reported.
아울러, 본 발명의 생들깨박 추출물을 유효 성분으로 포함하는 조성물은 상기 유효 성분 이외에 약제학적으로 적합하고 생리학적으로 허용되는 보조제를 사용하여 제조될 수 있으며, 상기 보조제로는 부형제, 붕해제, 감미제, 결합제, 피복제, 팽창제, 윤활제, 활택제 또는 향미제 등을 사용할 수 있다.In addition, the composition comprising the raw perilla plant extract of the present invention as an active ingredient may be prepared by using pharmaceutically acceptable and physiologically acceptable adjuvants in addition to the above-mentioned effective ingredients. Examples of the adjuvants include excipients, disintegrants, sweeteners , A binder, a coating agent, a swelling agent, a lubricant, a lubricant or a flavoring agent.
상기 조성물은 투여를 위해서 상기 기재한 유효 성분 이외에 추가로 약제학적으로 허용 가능한 담체를 1종 이상 포함하여 약제학적 조성물로 바람직하게 제제화할 수 있다.The composition may be formulated into a pharmaceutical composition containing at least one pharmaceutically acceptable carrier in addition to the above-described active ingredients for administration.
상기 약제학적 조성물의 제제 형태는 과립제, 산제, 정제, 피복정, 캡슐제, 좌제, 액제, 시럽, 즙, 현탁제, 유제, 점적제 또는 주사 가능한 액제 등이 될 수 있다. The pharmaceutical composition may be in the form of granules, powders, tablets, coated tablets, capsules, suppositories, liquids, syrups, juices, suspensions, emulsions, drops or injectable solutions.
예를 들어, 정제 또는 캡슐제의 형태로의 제제화를 위해,유효 성분은 에탄올, 글리세롤, 물 등과 같은 경구, 무독성의 약제학적으로 허용 가능한 불활성 담체와 결합될 수 있다. For example, for formulation into tablets or capsules, the active ingredient may be combined with an oral, non-toxic pharmaceutically acceptable inert carrier such as ethanol, glycerol, water, and the like.
또한, 원하거나 필요한 경우, 적합한 결합제, 윤활제, 붕해제 및 발색제 또한 혼합물로 포함될 수 있다. 적합한 결합제는 이에 제한되는 것은 아니나, 녹말, 젤라틴, 글루코스 또는 베타-락토오스와 같은 천연 당, 옥수수 감미제, 아카시아, 트래커캔스 또는 소듐올레이트와 같은 천연 및 합성 검, 소듐 스테아레이트, 마그네슘 스테아레이트, 소듐 벤조에이트, 소듐 아세테이트, 소듐 클로라이드 등을 포함하며, 녹말, 메틸 셀룰로스, 아가, 벤토니트, 잔탄 검 등을 포함한다. Also, if desired or necessary, suitable binders, lubricants, disintegrants and coloring agents may also be included as a mixture. Suitable binders include, but are not limited to, natural sugars such as starch, gelatin, glucose or beta-lactose, natural and synthetic gums such as corn sweeteners, acacia, tracker candles or sodium oleate, sodium stearate, magnesium stearate, sodium Benzoate, sodium acetate, sodium chloride and the like, and include starch, methyl cellulose, agar, bentonite, xanthan gum and the like.
액상 용액으로 제제화되는 조성물에 있어서 허용 가능한 약제학적 담체로는, 멸균 및 생체에 적합한 것으로서, 식염수, 멸균수, 링거액, 완충 식염수, 알부민 주사용액, 덱스트로즈 용액, 말토 덱스트린 용액, 글리세롤, 에탄올 및 이들 성분 중 1 성분 이상을 혼합하여 사용할 수 있으며, 필요에 따라 항산화제, 완충액, 정균제 등 다른 통상의 첨가제를 첨가할 수 있다. Acceptable pharmaceutical carriers for compositions that are formulated into a liquid solution include sterile solutions suitable for the living body such as saline, sterile water, Ringer's solution, buffered saline, albumin injection solution, dextrose solution, maltodextrin solution, glycerol, One or more of these components may be mixed and used. If necessary, other conventional additives such as an antioxidant, a buffer, and a bacteriostatic agent may be added.
또한, 희석제, 분산제, 계면활성제, 결합제 및 윤활제를 부가적으로 첨가하여 수용액, 현탁액, 유탁액 등과 같은 주사용 제형, 환약, 캡슐, 과립 또는 정제로 제제화 할 수 있다. 더 나아가 해당분야의 적절한 방법으로 Remington's Pharmaceutical Science, Mack Publishing Company, Easton PA에 개시되어 있는 방법을 이용하여 각 질환에 따라 또는 성분에 따라 바람직하게 제제화 할 수 있다. In addition, diluents, dispersants, surfactants, binders, and lubricants can be additionally added and formulated into injectable solutions, pills, capsules, granules or tablets such as aqueous solutions, suspensions, emulsions and the like. Further, it can be suitably formulated according to each disease or ingredient, using the method disclosed in Remington's Pharmaceutical Science, Mack Publishing Company, Easton PA as an appropriate method in the field.
또한, 본 발명은 상기 생들깨박 추출물을 포함하는 식품 조성물을 제공한다.본 발명에 따른 식품 조성물은 상기 약제학적 조성물과 동일한 방식으로 제제화 되어 기능성 식품으로 이용하거나, 각종 식품에 첨가할 수 있다. 본 발명의 조성물을 첨가할 수 있는 식품으로는 예를 들어, 음료류, 육류, 초코렛, 과자류, 피자, 라면, 면류, 껌류, 캔디, 젤리, 아이스크림류, 유제품, 알코올 음료류, 비타민 복합제, 건강보조 식품류 등이 있다. In addition, the present invention provides a food composition comprising the pericarpella extract. The food composition according to the present invention can be formulated in the same manner as the above pharmaceutical composition, and used as a functional food or added to various foods. Foods to which the composition of the present invention can be added include, for example, beverages, meat, chocolates, confectionery, pizza, ramen noodles, gums, candies, jellies, ice cream, dairy products, alcoholic beverages, .
본 발명은 또한 결핵 예방 및 치료용 의약 또는 식품의 제조를 위한 상기 생들깨박 추출물을 유효 성분으로 포함하는 조성물의 용도를 제공한다. 상기 생들깨박 추출물을 유효 성분으로 포함하는 본 발명의 조성물은 결핵 예방 및 치료와 관련된 질환의 예방 또는 치료용 의약 또는 식품의 제조 용도로 이용될 수 있다. The present invention also provides the use of a composition comprising said raw perilla oil extract as an active ingredient for the manufacture of medicaments or foods for the prevention and treatment of tuberculosis. The composition of the present invention comprising the raw perilla plant extract as an active ingredient can be used for the manufacture of medicines or foods for the prevention or treatment of diseases related to the prevention and treatment of tuberculosis.
또한, 본 발명은 치료상 유효량의 생들깨박 추출물을 투여하는 것을 포함하는 염증성 질환의 예방 또는 치료 방법을 제공한다. The present invention also provides a method of preventing or treating an inflammatory disease, comprising administering a therapeutically effective amount of fresh persimmon extract.
여기에서 사용된 용어 "치료상 유효량"은 연구자, 수의사, 의사 또는 기타 임상의에 의해 생각되는 조직계, 동물 또는 인간에서 생물학적 또는 의학적 반응을 유도하는 유효 성분 또는 약학적 조성물의 양을 의미하는 것으로, 이는 치료되는 질환 또는 장애의 증상의 완화를 유도하는 양을 포함한다. 본 발명의 유효 성분에 대한 치료상 유효 투여량 및 투여횟수는 원하는 효과에 따라 변화될 것임은 당업자에게 자명하다. 그러므로, 투여될 최적의 투여량은 당업자에 의해 쉽게 결정될 수 있으며, 질환의 종류, 질환의 중증도, 조성물에 함유된 유효성분 및 다른 성분의 함량, 제형의 종류, 및 환자의 연령, 체중, 일반 건강 상태, 성별 및 식이, 투여 시간, 투여 경로 및 조성물의 분비율, 치료기간, 동시 사용되는 약물을 비롯한 다양한 인자에 따라 조절될 수 있다. As used herein, the term "therapeutically effective amount " refers to the amount of active ingredient or pharmaceutical composition that induces a biological or medical response in a tissue system, animal or human, as contemplated by a researcher, veterinarian, This includes an amount that induces relief of the symptoms of the disease or disorder being treated. It will be apparent to those skilled in the art that the therapeutically effective dose and the number of administrations of the active ingredient of the present invention will vary depending on the desired effect. Thus, the optimal dosage to be administered can be readily determined by those skilled in the art and will vary with the nature of the disease, the severity of the disease, the amount of active and other ingredients contained in the composition, the type of formulation, and the age, The age, body weight, sex, diet, time of administration, route of administration and fraction of the composition, duration of treatment, concurrent medication, and the like.
본 발명에 있어서의 조성물 중 유효성분인 생들깨박 추출물의 투여량은 환자의 연령, 성별, 증상, 투여방법 또는 예방목적에 따라, 체중 kg 당 6 내지 30을 일일 1회 내지 3회 분복할 수 있다. 특이 증상을 나타내는 환자에 대한 투여용량 수준은 환자의 체중, 연령, 성별, 건강상태, 식이, 투여 시간, 투여 방법, 배설율, 질환의 중증도 등에 따라 당업자가 투여량을 변화시킬 수도 있다. The dose of the perilla perilla extract, which is an active ingredient in the composition of the present invention, may be 6 to 30 per kg of body weight per day, depending on the patient's age, sex, symptom, method of administration, have. Dosage levels for patients exhibiting the specific symptoms may vary depending on the patient's body weight, age, sex, health condition, diet, time of administration, administration method, excretion rate, severity of disease, and the like.
본 발명의 치료방법에서 본 발명의 생들깨박 추출물을 유효 성분으로 포함하는 조성물은 경구, 직장, 정맥내, 동맥내, 복강내, 근육내, 흉골내, 경피, 국소, 안구내 또는 피내 경로를 통해 통상적인 방식으로 투여할 수 있다. In the treatment method of the present invention, the composition comprising the raw perilla plant extract of the present invention as an active ingredient can be administered orally, rectally, intravenously, intraarterially, intraperitoneally, intramuscularly, intrasternally, transdermally, topically, ≪ / RTI > can be administered in a conventional manner.
[생들깨박 추출][Raw persimmon extract]
생들깨박은 충북음성 코메가에서 구입하여, 생들깨박 100 g을 1 L의 에탄올로 72시간동안 상온에서 추출하였다.The perilla seedlings were purchased from Chungbuk Song Komega and 100 g of raw perilla seeds were extracted with 1 L of ethanol at room temperature for 72 hours.
[항산화 시험][Antioxidant test]
시료인 생들깨박 추출물의 DPPH radical 소거 활성은 추출물 용액 30 μL와 에탄올에 용해한 60 μM DPPH 용액 30 μL를 각각 가하고, 상온에서 2분 동안 반응시켜 capillary tube에 옮긴 다음 electron spin resonance (ESR) spectrometer (JES-PX 2300, JEOL, Japan)로 측정하였다. The DPPH radical scavenging activity of the perilla perilla extract was determined by adding 30 μL of the extract solution and 30 μL of the 60 μM DPPH solution dissolved in ethanol. The reaction was carried out at room temperature for 2 minutes, transferred to a capillary tube, and then subjected to electron spin resonance (ESR) spectrometer JES-PX 2300, JEOL, Japan).
이 때 ESR spectrophotometer의 측정조건은 magnetic field의 경우 336.5mT, microwave power의 경우 5 mW, modulation frequency의 경우 9.41 GHz, modulation 항산화 시료에 대한 DPPH free radical 소거 활성 (%)은 (ESR signal intensity for medium containing the additives of concern/ESR signal intensity for the control medium) X100으로 계산한 결과, 10 μg/mL의 농도에서 65% 이상의 DPPH radical 소거활성을 나타내었고, 20 μg/mL의 농도에서는 80% 이상의 높은 DPPH radical 소거활성을 보이는 것을 확인하였다.([도 1]참조)At this time, the measurement conditions of the ESR spectrophotometer were 336.5mT for the magnetic field, 5mW for the microwave power, 9.41 GHz for the modulation frequency, and DPPH free radical scavenging activity (%) for the modulation antioxidant sample The DPPH radical scavenging activity was more than 65% at the concentration of 10 μg / mL and the DPPH radical scavenging activity was higher than 80% at the concentration of 20 μg / mL. (See Fig. 1). ≪ tb > < TABLE &
[세포생존율 시험][Cell survival rate test]
LPS로 자극된 BV-2세포에서 LPS 및 생들깨박 추출물이 세포 생존에 미치는 영향을 확인하기 위해 cell biability를 3-[4,5-Dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium bromide (MTT) 분석법으로 측정하였다. To determine the effect of LPS and perilla perilla extract on cell survival in BV-2 cells stimulated with LPS, cell biability was measured using 3- [4,5-Dimethylthiazol-2-yl] -2,5-diphenyl-tetrazolium bromide (MTT) assay.
세포(1X105 cell/ml)세포를 96-well plate에 180 μL씩 분주하여 12시간 이상 CO2 배양기에서 배양한 다음, 시료를 각각의 조건에 따라 처리하여 24시간 배양하였다. 배양한 후 배양액을 제거하고 0.5 mg/ml MTT가 함유되어 있는 배지 200 μL를 첨가한 다음 4시간 동안 배양하여 MTT가 환원되도록 하였다. 그 후 배양액을 제거하고 dimethylsulfoxide (DMSO) 100 μL 첨가하여 생성된 formazone결정을 용해시킨 후, ELISA reader를 이용하여 540 nm에서 흡광도를 측정하였다. 세포 생존율은 대조군과 비교하여 백분율(%)로 나타내었다. Cells (1 × 10 5 cells / ml) were seeded in a 96-well plate (180 μL), cultured in a CO2 incubator for more than 12 hours, and cultured for 24 hours. After culturing, the culture medium was removed, and 200 μL of the medium containing 0.5 mg / mL MTT was added, followed by incubation for 4 hours to reduce the MTT. After removing the culture medium, 100 μL of dimethylsulfoxide (DMSO) was added and the resulting formazone crystals were dissolved. The absorbance was measured at 540 nm using an ELISA reader. Cell viability was expressed as percentage (%) as compared with control.
Cell viability 측정 결과 LPS 및 생들깨박 추출물을 단독으로 또는 같이 처리한 모든 실험군에서 대조군에 비하여 세포 생존율이 변하지 않음을 확인하였다.([도 2] 참조)As a result of cell viability measurement, it was confirmed that the cell survival rate was not changed in all experimental groups treated with LPS and pericarp extract alone or in the same manner (as compared with the control group) (see FIG. 2).
이는 염증 유도 물질인 LPS와 생들깨박 추출물이 세포 생존에는 영향을 주지 않음을 나타낸다. This indicates that the inflammation inducers LPS and pericarp extract do not affect cell survival.
[LPS로 활성화된 신경교세포에서 생들깨박 추출물의 농도 의존적인 NO 생성저해 작용 시험][Concentration-dependent inhibition of NO production in fresh perilla plant extracts by LPS-activated glial cells]
생들깨박 추출물의 항염증 효능을 분석하기 위하여 본 발명에서는 염증 유발 인자인 LPS를 각 농도별로 자극된 신경교세포에서 생산되는 NO 농도를 의존적으로 효능을 있는지 확인하였다. In order to analyze the anti-inflammatory effect of the perilla plant extract, LPS, which is an inflammation inducer, was examined to determine whether or not the concentration of NO produced in the glial cells stimulated by each concentration was dependent on the efficacy.
NO 측정은 24 well plate에 세포를 5 x 105cell/을 seeding 한 후, LPS와 추출물을 농도차를 두어 첨가 한 후, 24h incubator에서 반응시킨 후, 각각 50 μL씩 Griess reagent (1% sulfanilamide/0.1% N-(1-naphthyl)- ethylenediamine dihydrochloride/2.5% H3PO4)와 반응 시킨 후, 파장이 540nm인 Sunrise basic 96-well microplate spectrophotometer (TECAN AUSTRIA)를 사용하여 값을 측정하였다.NO was determined by seeding 5 × 10 5 cells / well in a 24-well plate, adding LPS and extracts at different concentrations, reacting in a 24 h incubator, and then adding 50 μL each of Griess reagent (1% sulfanilamide / After reacting with 0.1% N- (1-naphthyl) -ethylenediamine dihydrochloride / 2.5% H3PO4), the values were measured using a Sunrise basic 96-well microplate spectrophotometer (TECAN AUSTRIA) with a wavelength of 540 nm.
신경교세포에서 LPS (1 ㎍/mL)에 의해서 유도되는 NO는 약 23 μM로 대조군으로 사용한 1 μM에 비하여 약 23배 이상 증가하였고, 실험군으로 생들깨박 추출물을 농도별로 처리를 한 결과 각각 5, 10, 25, 50 ㎍/mL로 NO 생성이 생들깨박 추출물의 농도 의존적으로 줄어드는 것을 확인하였다.([도 3] 참조)The concentration of NO induced by LPS (1 ㎍ / mL) in the glial cells was about 23 μM, which was about 23 times higher than that of 1 μM used in the control group. 10, 25, and 50 ㎍ / mL, the production of NO decreased depending on the concentration of the perilla seed extract (see FIG. 3).
[생들깨박 추출물 및 홍삼박 추출물][Perilla plant extracts and red ginseng leaves extract]
생들깨박과 홍삼박 각각 100 g을 1 L의 에탄올로 72시간동안 상온에서 추출하여 생들깨박 추출물 및 홍삼박 추출물을 제조하였다.100 g of fresh persimmon leaves and red ginseng leaves were extracted with 1 L of ethanol for 72 hours at room temperature to prepare fresh persimmon leaves and red ginseng leaves.
[항산화 시험][Antioxidant test]
시료인 생들깨박 및 홍삼박 추출물의 DPPH radical 소거 활성은 추출물 용액 30 μL와 에탄올에 용해한 60 μM DPPH 용액 30 μL를 각각 가하고, 상온에서 2분 동안 반응시켜 capillary tube에 옮긴 다음 electron spin resonance (ESR) spectrometer (JES-PX 2300, JEOL, Japan)로 측정하였다. The DPPH radical scavenging activity of each of the samples was investigated by adding 30 μL of the extract solution and 30 μL of 60 μM DPPH solution dissolved in ethanol. The reaction was carried out at room temperature for 2 minutes, transferred to a capillary tube, and then subjected to electron spin resonance ) spectrometer (JES-PX 2300, JEOL, Japan).
이때, 항산화 물질의 가장 특징적인 기작은 유리기와 반응하는 것으로 라디칼 소거 작용은 활성라디칼에 전자를 공여하여 항산화 효과나 인체에서 노화를 억제하는 척도로 이용되고 있다. 들깨박, 홍삼 복합 에탄올추출물의 라디칼 소거활성 측정은 DPPH 라디칼을 이용하여 측정하였다. DPPH는 짙은 자색을 띄는 비교적 안정한 활성 라디칼로서 항산화제, 방향족 아민류 등에 의해 환원되어 색이 탈색되는 것을 이용하여 항산화물질을 검색하였다. 따라서 들깨박, 홍삼 복합 에탄올추출물들과 천연 항산화제로 알려진 L-ascorbic acid의 항산화효과를 DPPH 라디칼의 소거활성을 측정하였고, DPPH 라디칼을 1/2로 소거하는데 필요한 시료의 농도인 RC50 값으로 나타내어 비교하여 [도 4]에 나타내었다. At this time, the most characteristic mechanism of the antioxidant is reacting with the free radical, and the radical scavenging function is used as an antioxidant effect by donating electrons to the active radical and a measure for suppressing the aging in the human body. The radical scavenging activity of perilla, red ginseng ethanol extract was measured by DPPH radical. DPPH is a relatively stable active radical with a deep violet color, which is reduced by antioxidants, aromatic amines, and the like. Therefore, the antioxidant activity of L-ascorbic acid, which is known as a natural antioxidant, was determined by measuring the scavenging activity of the DPPH radical and the RC 50 value, which is the concentration of the sample required to eliminate the DPPH radical, And is shown in Fig. 4 for comparison.
[도 4]에 나타난 결과, 들깨박, 홍삼박 복합 에탄올추출물의 혼합비율 중, 최적비율은 2:8 (홍삼추출물:들깨박추출물)의 비율이 가장 우수한 DPPH 라디칼 소거 활성을 보였다.As shown in FIG. 4, the optimum ratio of 2: 8 (red ginseng extract: perilla oil extract) to DPPH radical scavenging activity was the best among the mixture ratio of perilla and red ginseng mixed ethanol extracts.
이상의 설명은 본 발명의 기술사상을 예시적으로 설명한 것에 불과한 것으로서, 본 발명이 속하는 기술 분야에서 통상의 지식을 가진 자라면 본 발명의 본질적인 특성에서 벗어나지 않는 범위에서 다양한 수정 및 변형이 가능할 것이다. 따라서, 본 발명에 개시된 설명들은 본 발명의 기술 사상을 한정하기 위한 것이 아니라 설명하기 위한 것이고, 이러한 설명에 의하여 본 발명의 기술 사상의 범위가 한정되는 것은 아니다. 본 발명의 보호 범위는 아래의 청구범위에 의하여 해석되어야 하며, 그와 동등한 범위 내에 있는 모든 기술 사상은 본 발명의 권리범위에 포함되는 것으로 해석되어야 할 것이다.The foregoing description is merely illustrative of the technical idea of the present invention, and various changes and modifications may be made by those skilled in the art without departing from the essential characteristics of the present invention. Therefore, the description of the present invention is not intended to limit the scope of the present invention, but is to be construed as limited only by the scope of the present invention. The scope of protection of the present invention should be construed according to the following claims, and all technical ideas within the scope of equivalents should be construed as falling within the scope of the present invention.
Claims (18)
An antioxidative composition characterized by comprising a raw perilla oil extract as an active ingredient
상기 생들깨박 추출물을 유효성분으로 포함하는 항산화 조성물은 홍삼박 추출물을 더 포함하는 것을 특징으로 하는 항산화 조성물
The method according to claim 1,
Wherein the antioxidative composition comprising the raw persimmon leaf extract as an active ingredient further comprises an extract of red ginseng root.
상기 생들깨박 추출물 및 홍삼박 추출물은 생들깨박 에탄올 추출물 및 홍삼박 에탄올 추출물인 것을 특징으로 하는 항산화 조성물
3. The method of claim 2,
The antioxidative composition according to any one of the above items, wherein the raw perilla peach extract and the red ginseng root extract are the perilla perilla ethanol extract and the ethanol extract of red ginseng
상기 항산화 조성물은 항산화 및 항염증 활성 보조성분으로 석류추출액 및 프로폴리스를 더 포함하는 것을 특징으로 하는 항산화 조성물
3. The method of claim 2,
Wherein the antioxidant composition further comprises a pomegranate extract and propolis as an antioxidant and anti-inflammatory activity auxiliary component
An antiinflammatory composition comprising a perilla perilla extract as an active ingredient
상기 생들깨박 추출물을 유효성분으로 포함하는 항염증 조성물은 홍삼박 추출물을 더 포함하는 것을 특징으로 하는 항염증 조성물
6. The method of claim 5,
The anti-inflammatory composition comprising the perilla perilla extract as an active ingredient further comprises an extract of red ginseng root.
상기 생들깨박 추출물 및 홍삼박 추출물은 생들깨박 에탄올 추출물 및 홍삼박 에탄올 추출물인 것을 특징으로 하는 항염증 조성물
The method according to claim 6,
Wherein the raw perilla seed extract and the red ginseng root extract are raw perilla seed ethanol extracts and ethanol extracts of red ginseng seeds.
상기 항염증 조성물은 항산화 및 항염증 활성 보조성분으로 석류추출액 및 프로폴리스를 더 포함하는 것을 특징으로 하는 항염증 조성물
The method according to claim 6,
Wherein said anti-inflammatory composition further comprises a pomegranate extract and propolis as an antioxidant and anti-inflammatory activity adjunct ingredient.
상기 항염증 조성물은 유도성 질소 산화물 합성(iNOS) 및 NO 생산을 약화시키는 것을 특징으로 하는 항염증 조성물
The method of claim 5, wherein
Wherein said antiinflammatory composition is an antiinflammatory composition characterized by weakening inductive nitrogen oxide synthesis (iNOS) and NO production
상기 항염증 조성물은 미세아교세포에서 미세아교세포 활성을 억제하는 것을 특징으로 하는 항염증 조성물
6. The method of claim 5,
Wherein said anti-inflammatory composition inhibits microglial cell activity in microglial cells.
A composition for preventing or treating inflammatory neurodegenerative diseases, which comprises an extract of Perilla frutescens as an active ingredient
상기 생들깨박 추출물을 유효성분으로 포함하는 염증성 신경 퇴행성 질환 예방 또는 치료용 조성물은 홍삼박 추출물을 더 포함하는 것을 특징으로 하는 염증성 신경 퇴행성 질환 예방 또는 치료용 조성물
12. The method of claim 11,
The composition for preventing or treating an inflammatory neurodegenerative disease comprising the above-mentioned Perilla peach extract as an active ingredient further comprises a red ginseng extract. The composition for preventing or treating an inflammatory neurodegenerative disease
상기 생들깨박 추출물 및 홍삼박 추출물은 생들깨박 에탄올 추출물 및 홍삼박 에탄올 추출물인 것을 특징으로 하는 염증성 신경 퇴행성 질환 예방 또는 치료용 조성물
13. The method of claim 12,
The composition for preventing or treating inflammatory neurodegenerative diseases according to any one of the preceding claims, wherein the raw perilla peach extract and the red ginseng root extract are raw perilla seed ethanol extract and red ginseng seed ethanol extract
상기 염증성 신경 퇴행성 질환 예방 또는 치료용 조성물은 항산화 및 항염증 활성 보조성분으로 석류추출액 및 프로폴리스를 더 포함하는 것을 특징으로 하는 염증성 신경 퇴행성 질환 예방 또는 치료용 조성물
13. The method of claim 12,
Wherein the composition for preventing or treating inflammatory neurodegenerative diseases further comprises a pomegranate extract and propolis as an antioxidant and antiinflammatory activity auxiliary ingredient, a composition for preventing or treating inflammatory neurodegenerative diseases
상기 염증성 신경 퇴행성 질환 예방 또는 치료용 조성물은 유도성 질소 산화물 합성(iNOS) 및 NO 생산을 약화시키는 것을 특징으로 하는 염증성 신경 퇴행성 질환 예방 또는 치료용 조성물
12. The method of claim 11,
Wherein the composition for preventing or treating inflammatory neurodegenerative diseases is a composition for preventing or treating inflammatory neurodegenerative diseases, characterized by weakening inductive nitrogen oxide synthesis (iNOS) and NO production
상기 염증성 신경 퇴행성 질환 예방 또는 치료용 조성물은 미세아교세포에서 미세아교세포 활성을 억제하는 것을 특징으로 하는 염증성 신경 퇴행성 질환 예방 또는 치료용 조성물
12. The method of claim 11,
The composition for preventing or treating inflammatory neurodegenerative diseases is a composition for preventing or treating inflammatory neurodegenerative diseases characterized by inhibiting microglial cell activity in microglia
A food composition comprising a composition according to any one of claims 1 to 16
상기 식품 조성물은 음료, 육류, 초코렛, 과자류, 피자, 라면, 면류, 껌류, 캔디, 젤리, 아이스크림류, 유제품, 알코올 음료류, 비타민 복합제로부터 선택된 어느 하나인 것을 특징으로 하는 식품 조성물18. The method of claim 17,
Wherein the food composition is any one selected from beverage, meat, chocolate, confectionery, pizza, ramen, noodles, gum, candy, jelly, ice cream, milk product, alcoholic beverage,
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| KR100614970B1 (en) | 2004-08-05 | 2006-08-25 | 한국식품연구원 | Composition Comprising Defatted Perilla Extracted for Prevention and Attenuation of Cognitive Impairment |
| KR101153870B1 (en) | 2009-12-18 | 2012-07-19 | 대한민국 | Effective Perilla extract for inhibiting neuraminidase |
| KR101440372B1 (en) | 2013-05-30 | 2014-09-18 | 영농조합법인애농 | Food and pharmaceutical composition for anti-inflammation comprising extract of perilla sprout as effective component |
| KR20170024864A (en) | 2015-08-26 | 2017-03-08 | 백경연 | Method for manufacturing perilla oil by using perilla peels |
| KR101694660B1 (en) | 2016-09-27 | 2017-01-10 | 주식회사 더가든오브내추럴솔루션 | Ultrasonicating extract of Perilla frutescens buds under darkroom condition with anti-inflamatory effect |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20210055825A (en) * | 2019-11-07 | 2021-05-18 | 대한민국(농촌진흥청장) | Composition of extracts for antioxidant property containing silk worm pupae and red ginseng |
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