KR102527480B1 - Composition for preventing, improving or treating osteoporosis comprising camellia leaf extract - Google Patents
Composition for preventing, improving or treating osteoporosis comprising camellia leaf extract Download PDFInfo
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- KR102527480B1 KR102527480B1 KR1020220037928A KR20220037928A KR102527480B1 KR 102527480 B1 KR102527480 B1 KR 102527480B1 KR 1020220037928 A KR1020220037928 A KR 1020220037928A KR 20220037928 A KR20220037928 A KR 20220037928A KR 102527480 B1 KR102527480 B1 KR 102527480B1
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- 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)
- A61K36/82—Theaceae (Tea family), e.g. camellia
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- 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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- A23V2200/00—Function of food ingredients
- A23V2200/30—Foods, ingredients or supplements having a functional effect on health
- A23V2200/306—Foods, ingredients or supplements having a functional effect on health having an effect on bone mass, e.g. osteoporosis prevention
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Abstract
Description
본 발명은 골다공증의 예방, 개선 또는 치료용 조성물에 관한 것으로, 구체적으로는 동백 잎 추출물을 포함하는 골다공증의 예방, 개선 또는 치료용 조성물 및 상기 조성물의 제조방법에 관한 것이다.The present invention relates to a composition for preventing, improving or treating osteoporosis, and more specifically, to a composition for preventing, improving or treating osteoporosis comprising a camellia leaf extract and a method for preparing the composition.
동백나무는 차나무와 동일 속으로 지금까지 주로 관상용 같은 원예자원으로 이용되어 왔고, 부위별로는 고품질 지방이 함유되어 있는 종실이 전통적으로 식용유와 화장유 자원으로 이용되어 왔으며, 줄기는 고급 숯의 원료로 활용되어 왔으나 아직까지 식용 및 약용자원으로 이용되고 있지 않고 일부 연구만 진행되고 있는 실정이다. 동백나무는 우리나라의 울릉도, 제주도 및 남부 지방의 바닷가나 섬에서 자라며 개화기는 12월~5월이고 결실기는 9월~10월이다. 약재로 쓰이는 말린 동백꽃은 유연하고 좋은 향기가 나는데, 색이 붉고 곰팡이가 피지 않았으며 꽃봉오리가 크고 아직 피지않은 것이 제일 높은 등급을 갖는다. 산다화의 맛과 성질은 달고 약간 매우며 서늘하다. 동백꽃의 효능으로는 간경, 폐경에 들어가며 양혈, 지혈, 산어혈에 효과가 있다고 알려져 있다.Camellia, the same genus as tea, has been mainly used as a horticultural resource such as ornamental. By part, the seeds containing high-quality fat have traditionally been used as a resource for cooking oil and cosmetic oil, and the stem is used as a raw material for high-quality charcoal. However, it has not yet been used as a food or medicinal resource, and only some research is being conducted. Camellia grows in the beaches and islands of Ulleungdo, Jeju Island, and southern regions of Korea. The flowering period is December to May, and the fruiting period is September to October. Dried camellia flowers used as medicine are flexible and have a good scent. Those with red color, no mold, and large buds that have not yet bloomed have the highest grade. The taste and properties of sandalwood are sweet, slightly spicy and cool. As for the efficacy of camellia flower, it is known that it enters liver cirrhosis and menopause, and is effective for yang blood, hemostasis, and acid fish blood.
한편, 골다공증은 그 증세 자체보다는 골의 약화에 따라 용이하게 초래되는 각종 골절, 특히 대퇴골 골절 또는 척추골절 등이 장기간 활동을 제한하여 건강한 생활을 영위할 수 없고, 결과적으로 노인층 사망의 15%에 대한 원인제공을 하는 것으로 알려져 있다. 골량은 유전적 요인, 영양 섭취, 호르몬의 변화, 운동 및 생활 습관의 차이 등 여러 가지 요인들에 의해 영향을 받으며, 골다공증의 원인으로는 노령, 운동 부족, 저체중, 흡연, 저칼슘 식이, 폐경, 난소 절제 등이 알려져 있다.On the other hand, osteoporosis, rather than the symptom itself, is easily caused by bone weakness, and various fractures, especially femur fractures or vertebral fractures, restrict activity for a long time and cannot lead a healthy life, resulting in about 15% of deaths among the elderly. It is known to provide a cause. Bone mass is influenced by many factors, including genetic factors, nutritional intake, hormonal changes, and differences in exercise and lifestyle. Ovariectomy and the like are known.
개인 차이는 있지만 백인보다는 흑인이 골 재흡수 수준(bone resorption level)이 낮아 골량이 더 높으며, 대개 골량은 14~18세에 가장 높고 노후에는 1 년에 약 1 %씩 감소한다. 특히 여성의 경우 30 세 이후부터 골감소가 지속적으로 진행되며, 폐경기에 이르면 호르몬 변화에 의해 골 감소가 급격히 진행된다. 이와 같이 골다공증은 정도에 차이는 있으나 노년층, 특히 폐경기 이후의 여성에게 있어서는 피할 수 없는 증상으로, 선진국에서는 인구가 노령화됨에 따라 골다공증 및 그 치료제에 대한 관심이 점차 증가되고 있으나 동백나무, 동백꽃, 동백 잎 등을 이용한 골다골증의 예방 또는 치료에 대한 연구는 전무한 상태이다. Although there are individual differences, blacks have higher bone mass than whites because their bone resorption level is lower. Bone mass usually peaks between the ages of 14 and 18 and decreases by about 1% per year in old age. In particular, in the case of women, bone loss continues to progress after the age of 30, and bone loss rapidly progresses due to hormonal changes when reaching menopause. As described above, osteoporosis is an unavoidable symptom in the elderly, especially postmenopausal women, although the degree varies. As the population ages in developed countries, interest in osteoporosis and its treatment is gradually increasing, but camellia, camellia flower, and camellia leaf There is no study on the prevention or treatment of osteoporosis using the back.
상기와 같은 문제점을 해결하기 위해 본 발명은 동백 잎 추출물을 함유하는 골다공증 예방, 개선 또는 치료용 조성물을 제공하고자 한다.In order to solve the above problems, the present invention is to provide a composition for preventing, improving or treating osteoporosis containing a camellia leaf extract.
또한, 상기의 동백 잎 추출물의 제조하는 방법 및 동백 잎 추출물을 함유하는 골다공증 예방, 개선 또는 치료용 조성물의 제조방법을 제공하고자 한다.In addition, it is intended to provide a method for preparing the camellia leaf extract and a method for preparing a composition for preventing, improving or treating osteoporosis containing the camellia leaf extract.
상기와 같은 목적을 달성하기 위해 본 발명은 동백 잎 추출물을 유효성분으로 포함하는 골다공증의 예방, 개선, 치료 효과가 우수한 조성물을 제공한다.In order to achieve the above object, the present invention provides a composition containing a camellia leaf extract as an active ingredient and excellent in preventing, improving, and treating osteoporosis.
본 발명에 따른 동백 잎 추출물은 다음의 단계를 포함하는 제조 방법을 통해 제조될 수 있다: The camellia leaf extract according to the present invention can be prepared through a manufacturing method comprising the following steps:
(a) 동백 잎 준비 단계;(a) Camellia leaf preparation step;
(b) 건조된 동백 잎을 전처리하는 단계;(b) pre-treating the dried camellia leaves;
(c) 전처리된 동백 잎을 추출하는 단계. (c) extracting the pretreated camellia leaves.
상기 (a) 동백 잎 준비 단계는 추출에 사용할 동백 잎을 준비하는 단계로, 상기 동백 잎은 생신엽 또는 건신엽을 사용할 수 있다. 건신엽을 사용할 경우, 25 내지 35℃, 바람직하게는 30℃에서 2 내지 7일, 바람직하게는 3 내지 5일간 건조시켜준다. 상기 온도 및 기간 범위 미만이면 동백 잎이 제대로 건조되지 않고, 상기 온도 및 기간을 초과하면 잎이 너무 건조되어 이후의 찌는 단계, 덖음 단계에서 잎이 부서질 수 있다.The (a) camellia leaf preparation step is a step of preparing camellia leaves to be used for extraction, and fresh or dried leaves may be used as the camellia leaves. When dried leaves are used, they are dried at 25 to 35°C, preferably at 30°C for 2 to 7 days, preferably 3 to 5 days. If the temperature and period are less than the above ranges, the camellia leaves are not dried properly, and if the temperature and period are exceeded, the leaves are too dry, and the leaves may be broken in the subsequent steaming and roasting steps.
상기 동백 잎은 제한되는 것은 아니며, 성엽 또는 어린 잎을 사용할 수 있다. 어린 잎을 사용하는 경우 바람직하게는 1cm 이상 18cm 미만의 어린 잎일 수 있으며, 어린 잎을 사용할 때 골다공증의 예방, 개선 또는 치료 효과가 더욱 증대될 수 있다.The camellia leaves are not limited, and adult leaves or young leaves may be used. When using a young leaf, it may preferably be a young leaf of 1 cm or more and less than 18 cm, and when using a young leaf, the effect of preventing, improving or treating osteoporosis may be further increased.
상기 (b) 단계는 준비된 동백 잎을 비벼주는 유념, 덖고 비벼주는 덖음, 잎을 찌는 찜, 찌고 덖고 비벼주는 찜/덖음 처리 중 어느 하나 이상을 수행하여 전처리하는 단계이다. 덖음하는 단계는 180 내지 260℃, 바람직하게는 200 내지 240℃에서 1 내지 10분간, 바람직하게는 3 내지 5분간 덖음하고, 덖음된 동백 잎을 상온(20 내지 25℃)에서 3 내지 5분간 식혀준다. 이때, 햇빛이 들지 않는 그늘에서 식혀줘야 한다.Step (b) is a step of pre-processing the prepared camellia leaves by performing one or more of the following: mindfulness of rubbing, roasting and rubbing, steaming of leaves, and steaming/steaming of steaming and rubbing. The roasting step is 180 to 260 ° C, preferably 200 to 240 ° C for 1 to 10 minutes, preferably 3 to 5 minutes, and the boiled camellia leaves are cooled at room temperature (20 to 25 ° C) for 3 to 5 minutes. give. At this time, it should be cooled in the shade without sunlight.
동백 잎을 찌는 단계는 90 내지 110℃, 바람직하게는 100℃에서 10 내지 120분, 바람직하게는 30 내지 60분간 쪄준다. 상기 온도 범위 및 시간을 벗어나는 경우 골다공증의 예방, 개선 또는 치료 효과가 저하될 수 있다.The step of steaming the camellia leaves is steamed at 90 to 110 ° C, preferably 100 ° C for 10 to 120 minutes, preferably 30 to 60 minutes. Outside the above temperature range and time, the effect of preventing, improving or treating osteoporosis may be reduced.
찜/덖음 처리는 상기 찜 처리, 덖음 처리 조건으로 각각 수행될 수 있다.The steaming/roasting process may be performed under the conditions of the steaming process and the roasting process, respectively.
상기 (c) 단계는 (b) 단계에서 전처리된 동백 잎을 추출하여 동백 잎 추출물을 수득하는 단계이다.Step (c) is a step of obtaining a camellia leaf extract by extracting the camellia leaves pretreated in step (b).
상기 추출물에 사용되는 용매는 제한되는 것은 아니지만 바람직하게는 물 또는 정제수를 용매로 하여 추출될 수 있다.The solvent used for the extract is not limited, but may preferably be extracted using water or purified water as a solvent.
또한, 상기 추출물은 동백 잎을 60 내지 80℃에서 3 내지 10분, 바람직하게는 5 내지 10분, 더욱 바람직하게는 5분 간 추출한 것일 수 있다. 상기 온도 및 시간 외의 조건으로 추출될 경우 유효성분이 충분하게 추출되지 않을 수 있다.In addition, the extract may be obtained by extracting camellia leaves at 60 to 80° C. for 3 to 10 minutes, preferably 5 to 10 minutes, and more preferably 5 minutes. When extracted under conditions other than the above temperature and time, active ingredients may not be sufficiently extracted.
본 발명은 또한, 동백 잎 추출물을 포함하는 골다공증의 예방, 개선 또는 치료용 조성물을 제공한다.The present invention also provides a composition for preventing, improving or treating osteoporosis comprising a camellia leaf extract.
상기 골다공증의 예방, 개선 또는 치료용 조성물은 약학적 조성물로 제공될 수 있다.The composition for preventing, improving or treating osteoporosis may be provided as a pharmaceutical composition.
또는 상기 조성물이 식품 조성물로 제공되는 경우, 골다공증의 예방 또는 개선용 식품 조성물로 제공된다.Alternatively, when the composition is provided as a food composition, it is provided as a food composition for preventing or improving osteoporosis.
본 발명에서의 약학적 조성물 또는 식품 조성물은 본 발명에 따른 동백 잎 추출물은 0 초과, 바람직하게는 10ug/mL 이상의 농도로 포함될 수 있으나, 이에 제한되는 것은 아니다.In the pharmaceutical composition or food composition of the present invention, the camellia leaf extract according to the present invention may be included in a concentration of greater than 0, preferably 10ug/mL or more, but is not limited thereto.
또한, 상기 약학적 조성물은 약학적으로 허용되는 담체, 결합제, 활탁제, 붕해제, 부형제, 가용화제, 분산제, 안정화제, 현탁화제 등을 추가적으로 더 포함할 수 있고, 예를 들면 락토즈, 덱스트로즈, 수크로즈, 솔비톨, 만니톨, 자일리톨, 에리스리톨, 말디톨, 전분, 알지네이트, 칼슘 포스페이트, 칼슘 실리케이트, 말토덱스트린, 이산화규소, 셀룰로오스, 메틸 셀룰로오스, 미정질 셀룰로오스, 폴리비닐피롤리돈, 메틸하이드록시벤조에이트, 프로필하이드록시벤조에이트, 탈크, 마그네슘 스테아레이트 등일 수 있으나 이에 제한되는 것은 아니다.In addition, the pharmaceutical composition may further include a pharmaceutically acceptable carrier, binder, lubricant, disintegrant, excipient, solubilizer, dispersant, stabilizer, suspending agent, etc., for example, lactose, dex Trose, sucrose, sorbitol, mannitol, xylitol, erythritol, malditol, starch, alginate, calcium phosphate, calcium silicate, maltodextrin, silicon dioxide, cellulose, methyl cellulose, microcrystalline cellulose, polyvinylpyrrolidone, methylhydride It may be hydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, etc., but is not limited thereto.
또한, 상기 약학 조성물은 당 업계에서 제제화에 통상적으로 사용되는 방법으로 정제, 캡슐, 서방형 제제, 엘릭서, 서스펜션, 시럽, 웨이퍼, 단위 투약 주사제 앰플, 현탁액 등의 제형으로 제조될 수 있다.In addition, the pharmaceutical composition may be prepared in a formulation such as a tablet, capsule, sustained-release preparation, elixir, suspension, syrup, wafer, unit dose injection ampoule, suspension, etc. by a method commonly used for formulation in the art.
상기 약학 조성물의 투여 방식은 경구 투여, 정맥내 투여, 근육내 투여, 동맥내 투여, 골수내 투여, 경막내 투여, 심장내 투여, 경피 투여, 피하 투여, 복강내 투여, 비강내 투여, 장관 투여, 구강 및 설하는 포함하는 국소 투여, 직장 투여, 국소 투여, 진피 투여, 점막 투여, 척추내 투여 등의 방법으로 투여될 수 있고, 바람직하게는 경구 투여일 수 있으나, 이에 제한되는 것은 아니다. 또한, 상기 투여 시 각 투여 방식에 절절한 제형으로 제제화되어 투여될 수 있으며, 제제화는 당 업계에서 널리 알려져 있는 방법으로 제제화될 수 있다.The administration method of the pharmaceutical composition is oral administration, intravenous administration, intramuscular administration, intraarterial administration, intramedullary administration, intrathecal administration, intracardiac administration, transdermal administration, subcutaneous administration, intraperitoneal administration, intranasal administration, enteral administration , It may be administered by methods such as topical administration, rectal administration, topical administration, dermal administration, mucosal administration, intravertebral administration, including oral and sublingual administration, preferably oral administration, but is not limited thereto. In addition, at the time of administration, it may be formulated and administered in a formulation suitable for each administration method, and the formulation may be formulated by a method widely known in the art.
상기 약학 조성물은 투여 대상의 연령, 체중, 건강 상태, 성별, 투여시간, 투여 경로, 배출율, 유효성분의 함량, 예방 또는 치료될 특정 질환의 중증도 등에 따라 적절하게 선택될 수 있고, 투여 횟수는 1일 1회 또는 2회, 격일마다, 격주마다, 격월 등의 방식으로 투여할 수 있다.The pharmaceutical composition may be appropriately selected according to the age, weight, health condition, sex, administration time, administration route, excretion rate, active ingredient content, severity of a specific disease to be prevented or treated, etc., and the number of administrations is 1 It may be administered once or twice a day, every other day, every other week, or every other month.
또한, 상기 식품 조성물은 당 업계에서 식품을 제조하기 위해 통상적으로 사용하는 향미제, 풍미제, 안정화제, 증점제, 방부제 등을 추가적으로 더 포함할 수 있고, 예를 들면 락토즈, 덱스트로즈, 수크로즈, 솔비톨, 만니톨, 자일리톨, 에리스리톨, 말디톨, 전분, 알지네이트, 칼슘 포스페이트, 칼슘 실리케이트, 말토덱스트린, 이산화규소, 셀룰로오스, 메틸 셀룰로오스, 미정질 셀룰로오스, 폴리비닐피롤리돈, 메틸하이드록시벤조에이트, 프로필하이드록시벤조에이트, 탈크, 마그네슘 스테아레이트 등일 수 있으나 이에 제한되는 것은 아니다.In addition, the food composition may additionally include flavoring agents, flavoring agents, stabilizers, thickeners, preservatives, etc. commonly used in the art to prepare food, for example, lactose, dextrose, water chlorose, sorbitol, mannitol, xylitol, erythritol, malditol, starch, alginate, calcium phosphate, calcium silicate, maltodextrin, silicon dioxide, cellulose, methyl cellulose, microcrystalline cellulose, polyvinylpyrrolidone, methylhydroxybenzoate, It may be propyl hydroxybenzoate, talc, magnesium stearate, etc., but is not limited thereto.
상기 식품 조성물은 음료, 껌, 차, 비타민 복합제, 분말, 과립, 정제, 캡슐,과자, 등의 형태로 제조될 수 있다. The food composition may be prepared in the form of beverages, gum, tea, vitamin complexes, powders, granules, tablets, capsules, confectionery, and the like.
상기 식품 조성물은 섭취 대상의 연령, 체중, 건강 상태, 성별, 투여시간, 유효성분의 함량, 질환의 중증도 등에 따라 적절하게 선택될 수 있고, 섭취 횟수는 1일 1회 또는 2회, 격일마다, 격주마다, 격월 등의 방식으로 섭취할 수 있다.The food composition may be appropriately selected according to the age, weight, health condition, sex, administration time, content of the active ingredient, severity of the disease, etc. of the subject to be consumed, and the number of intakes is once or twice a day, every other day, You can take it every other week, every other month, etc.
본 발명은 동백 잎 추출물을 포함함으로써 골다골증의 예방, 개선 또는 치료 효과가 우수한 조성물을 제공할 수 있으며, 상기 동백 잎 추출물의 제조방법 및 동백 잎 추출물을 포함하는 골다공증의 예방, 개선 또는 치료용 조성물의 제조방법을 제공할 수 있다.The present invention can provide a composition containing a camellia leaf extract that has excellent effects on preventing, improving or treating osteoporosis, and the method for preparing the camellia leaf extract and the composition for preventing, improving or treating osteoporosis containing the camellia leaf extract A manufacturing method can be provided.
도 1은 본 발명에 따라 제조된 어린 동백 잎 추출물의 농도에 따른 조골세포 분화 개선 효능 평가 결과를 나타낸 것이다.
도 2는 본 발명에 따라 제조된 어린 동백 잎 추출물의 농도에 따른 조골세포 분화 개선 효능 평가 결과를 나타낸 것이다.
도 3은 본 발명에 따라 제조된 동백 성엽 추출물의 농도에 따른 조골세포 분화 개선 효능 평가 결과를 나타낸 것이다.
도 4는 본 발명에 따라 제조된 어린 동백 찜/덖음 잎의 추출물의 시간에 따른 조골세포 분화 개선 효능 평가 결과를 나타낸 것이다.1 shows the results of evaluating the osteoblast differentiation improvement efficacy according to the concentration of the young camellia leaf extract prepared according to the present invention.
Figure 2 shows the results of osteoblast differentiation improvement efficacy evaluation according to the concentration of the young camellia leaf extract prepared according to the present invention.
Figure 3 shows the osteoblast differentiation improvement efficacy evaluation results according to the concentration of the camellia seongyeop extract prepared according to the present invention.
Figure 4 shows the results of the evaluation of the osteoblast differentiation improvement efficacy over time of the extract of young camellia steamed / boiled leaves prepared according to the present invention.
이하, 본 발명을 실시예 및 실험예에 의해 상세히 설명한다.Hereinafter, the present invention will be described in detail by examples and experimental examples.
단, 하기 실시예 및 실험예는 본 발명을 예시하는 것일 뿐, 본 발명의 내용이 하기 실시예 및 실험예에 한정되는 것은 아니다.However, the following Examples and Experimental Examples are merely illustrative of the present invention, and the contents of the present invention are not limited to the following Examples and Experimental Examples.
<실시예 1 내지 3> 본 발명에 따른 생 동백 잎 추출물 제조<Examples 1 to 3> Preparation of fresh camellia leaf extract according to the present invention
동백 잎 차를 제조하기 위해 생잎을 사용하였고, 사용된 동백 잎 중에서 어린잎은 새로 자란 18cm미만을 기준으로 채취하였다. 생엽(생신엽)은 덖고 비벼주는 조건 (덖음), 찌기만한 조건 (찜), 찌고 덖어주고 비벼주는 조건 (찜/덖음)으로 각각 제조하였다. 유념은 동백 잎을 면보에 쌓아서 비벼주었고, 덖음은 200℃~230℃정도에서 3분간 덖어주고 식혀주기를 2회 반복하였다. 찜은 100℃에서 30분간 쪄주고 건조하였고, 찜/덖음은 100℃에서 30분간 찌고 식힌 다음 200℃~230℃정도에서 3분간 덖어주고 식혀주기를 2회 반복하였다.Fresh leaves were used to prepare camellia leaf tea, and among the used camellia leaves, young leaves were collected based on newly grown less than 18 cm. Saengyeop (saengsinyeop) was prepared under the conditions of roasting and rubbing (steaming), steaming conditions (steaming), and steaming and rubbing conditions (steaming/steaming). For mindfulness, camellia leaves were stacked on a cotton cloth and rubbed, and for 3 minutes at 200℃~230℃ for 3 minutes, the cooling cycle was repeated twice. The steam was steamed at 100 ° C for 30 minutes and dried, and the steam / roast was steamed at 100 ° C for 30 minutes, cooled, then steamed at 200 ° C to 230 ° C for 3 minutes, and the cooling cycle was repeated twice.
상기 과정에 의해 얻어진 각각의 동백 잎 1g을 차우림백에 넣고 약 60℃~80℃ 물에 5분간 담궈 우린 다음 차우림백을 제거하였다. 동백 잎 추출물을 원심분리 후 상층액을 0.45μm 주사기 필터를 이용하여 여과하고 -70℃ 초저온냉동고에서 24시간 보관한 다음 동결 건조하였다. 동결 건조된 샘플은 상온에 보관하였다.1 g of each camellia leaf obtained by the above process was put in a tea rim bag, soaked in water at about 60 ° C to 80 ° C for 5 minutes, and then the tea rim bag was removed. After the camellia leaf extract was centrifuged, the supernatant was filtered using a 0.45 μm syringe filter, stored in a -70 ° C ultra-low temperature freezer for 24 hours, and then freeze-dried. Freeze-dried samples were stored at room temperature.
<실시예 4 내지 7> 본 발명에 따른 건조 동백 잎 추출물 제조<Examples 4 to 7> Preparation of dried camellia leaf extract according to the present invention
본 발명에 따른 건조된 동백 잎 추출물 제조 새로 자란 18cm미만을 기준의 어린 동백 잎을 채취하고, 건조를 위해 세척한 다음 30℃에서 5일간 건조하였다. Preparation of dried camellia leaf extract according to the present invention Newly grown young camellia leaves of less than 18 cm were collected, washed for drying, and then dried at 30 ° C. for 5 days.
실시예 1 내지 3과 마찬가지로, 건조된 신엽을 각각 비벼주기만 한 조건 (유념), 덖고 비벼주는 조건 (덖음), 찌기만한 조건 (찜), 찌고 덖어주고 비벼주는 조건 (찜/덖음)으로 제조하였다. 유념은 동백 잎을 면보에 쌓아서 비벼주었고, 덖음은 200℃~230℃정도에서 3분간 덖어주고 식혀주기를 2회 반복하였다. 찜은 100℃에서 30분간 쪄주고 건조하였고, 찜/덖음은 100℃에서 30분간 찌고 식힌 다음 200℃~230℃정도에서 3분간 덖어주고 식혀주기를 2회 반복하였다.As in Examples 1 to 3, the dried new leaves were prepared under the condition of only rubbing (mind), the condition of rubbing and rubbing (steaming), the condition of steaming (steaming), and the condition of steaming, rubbing and rubbing (steaming/steaming). . For mindfulness, camellia leaves were stacked on a cotton cloth and rubbed, and for 3 minutes at 200℃~230℃ for 3 minutes, the cooling cycle was repeated twice. The steam was steamed at 100 ° C for 30 minutes and dried, and the steam / roast was steamed at 100 ° C for 30 minutes, cooled, then steamed at 200 ° C to 230 ° C for 3 minutes, and the cooling cycle was repeated twice.
상기 과정에 의해 얻어진 각각의 동백 잎 1g을 차우림백에 넣고 약 60~80℃ 물에 5분간 담궈 우린 다음 차우림백을 제거하였다. 동백 잎 추출물을 원심분리 후 상층액을 0.45μm 주사기 필터를 이용하여 여과하고 -70℃ 초저온냉동고에서 24시간 보관한 다음 동결 건조하였다. 동결 건조된 샘플은 상온에 보관하였다.1 g of each camellia leaf obtained by the above process was put in a tea rinsing bag, soaked in water at about 60 to 80 ° C for 5 minutes, and then the tea rinsing bag was removed. After the camellia leaf extract was centrifuged, the supernatant was filtered using a 0.45 μm syringe filter, stored in a -70 ° C ultra-low temperature freezer for 24 hours, and then freeze-dried. Freeze-dried samples were stored at room temperature.
<실시예 8 내지 10> 본 발명에 따른 동백 성엽 추출물 제조<Examples 8 to 10> Preparation of camellia sage leaf extract according to the present invention
상기 실시예 1 내지 3과 동일하게 제조하되, 사용된 동백 잎은 어린 잎이 아닌 성엽을 사용하였다.It was prepared in the same manner as in Examples 1 to 3, but the used camellia leaves were adult leaves, not young leaves.
<실시예 11 내지 14> 본 발명에 따른 건조된 동백 성엽 추출물 제조 2<Examples 11 to 14> Preparation 2 of dried camellia sex leaf extract according to the present invention
상기 실시예 4 내지 7과 동일한 방법으로 제조하되, 사용된 동백 잎은 어린 잎이 아닌 5월에 채취한 성엽을 사용하였다.It was prepared in the same way as in Examples 4 to 7, but the camellia leaves used were adult leaves collected in May, not young leaves.
<실시예 15 내지 18> 본 발명에 따른 건조된 동백 성엽 추출물 제조 3<Examples 15 to 18>
상기 실시예 4 내지 7과 동일한 방법으로 제조하되, 사용된 동백 잎은 어린 잎이 아닌 9월에 채취한 성엽을 사용하였다.It was prepared in the same way as in Examples 4 to 7, but the camellia leaves used were adult leaves collected in September, not young leaves.
<실험예 1> 동백 어린잎 차의 농도에 따른 조골세포 분화 개선 효과 평가 1<Experimental Example 1> Evaluation of the osteoblast differentiation improvement effect according to the concentration of camellia young leaf tea 1
실시예 1 내지 7의 추출물의 농도를 각각 10 μg/mL 및 20 μg/mL으로 설정하고, 이를 이용하여 조골세포 분화 개선 효능을 평가하였다. The concentrations of the extracts of Examples 1 to 7 were set to 10 μg/mL and 20 μg/mL, respectively, and the efficacy of improving osteoblast differentiation was evaluated using this.
조골세포 분화 정도를 분석하기 위해서 ALP 염색 키트인 TRACP & ALP double-stain Kit (MK300, TaKaRa)를 이용하였다. 6일 후 배양액을 제거하고 PBS로 씻어준 다음 고정액 200μl를 각 well에 넣어주고 5분간 반응시켜 고정해주었다. 2ml의 멸균수를 이용하여 2번 씻어준 다음 ALP 기질을 200μl 넣어주고 37℃에서 30분간 반응시켰다.In order to analyze the degree of osteoblast differentiation, an ALP staining kit, TRACP & ALP double-stain kit (MK300, TaKaRa) was used. After 6 days, the culture medium was removed, washed with PBS, and 200 μl of the fixative was added to each well, followed by reaction for 5 minutes to fix. After washing twice with 2ml of sterile water, 200μl of ALP substrate was added and reacted at 37℃ for 30 minutes.
분화 6일 후의 평가 결과는 도 1에 나타내었고, 염색이 진하게 될수록 ALP 활성이 높고 조골세포로 분화가 잘 된 것이다.The evaluation results after 6 days of differentiation are shown in FIG. 1, and the darker the staining, the higher the ALP activity and the better the differentiation into osteoblasts.
평가 결과, 조골 유사 세포인 MG-63세포를 분화 유도하고 동백 어린잎 추출물을 각각 처리했을 때 추출물 처리시 분화 배양액에 비해 조골세포 분화가 더욱 잘 되는 것을 확인할 수 있었다. 특히 찜/덖음 처리 동백 잎 추출물이 다른 추출물에 비해 가장 분화를 잘 유도했다. 이러한 결과를 통해 동백 잎 추출물은 골다공증 예방, 개선 또는 치료가 가능하다고 판단된다.As a result of the evaluation, it was confirmed that when MG-63 cells, which are osteoblast-like cells, were differentiated and treated with the camellia young leaf extract, respectively, osteoblast differentiation was better than that of the differentiation culture medium when the extract was treated. In particular, the steamed/roasted camellia leaf extract induced differentiation the best compared to other extracts. Through these results, it is judged that the camellia leaf extract can prevent, improve or treat osteoporosis.
<실험예 2> 동백 어린잎 차의 농도에 따른 조골세포 분화 개선 효과 평가 2<Experimental Example 2> Evaluation of the osteoblast differentiation improvement effect according to the concentration of camellia young leaf tea 2
실시예 4 내지 7의 추출물의 농도를 각각 10 μg/mL 및 20 μg/mL으로 설정하고, 이를 이용하여 조골세포 분화 개선 효능을 평가하였다. Concentrations of the extracts of Examples 4 to 7 were set to 10 μg/mL and 20 μg/mL, respectively, and the efficacy of improving osteoblast differentiation was evaluated using this.
조골세포 분화 정도를 분석하기 위해 ALP 활성을 TRACP & ALP Assay Kit (MK301, TaKaRa)를 이용하여 진행하였다. 6일 후 배양액을 제거하고 PBS로 씻어준 다음 세포 용해액 50μl를 각 well에 넣어주고 세포를 용해시켰다. 세포 용해액 40μl에 p-nitro-phenyl phosphate 기질 40μl를 넣어주고 37℃에서 1시간 반응시킨 후 0.5N NaOH 40μl를 넣어 반응을 정지시키고 흡광도 405nm에서 측정하였다.In order to analyze the degree of osteoblast differentiation, ALP activity was performed using TRACP & ALP Assay Kit (MK301, TaKaRa). After 6 days, the culture medium was removed, washed with PBS, and 50 μl of the cell lysate was added to each well to lyse the cells. After adding 40 μl of p -nitro-phenyl phosphate substrate to 40 μl of the cell lysate and reacting at 37° C. for 1 hour, 40 μl of 0.5N NaOH was added to stop the reaction, and the absorbance was measured at 405 nm.
분화 6일 후의 평가 결과는 도 2에 나타내었고, * p<0.05, ** p<0.01 이다.The evaluation results after 6 days of differentiation are shown in Figure 2, * p <0.05, ** p <0.01.
평가 결과, 조골 유사 세포인 MG-63세포를 분화 유도하고 동백 어린잎 추출물을 각각 처리했을 때 추출물 처리시 분화 배양액에 비해 ALP 활성이 높게 나타는 것으로 보아 조골세포 분화가 더욱 잘 되는 것을 확인할 수 있었다. 하지만 찜/덖음 처리 추출물에서 ALP염색이 가장 진하게 된 것과는 다르게 ALP 활성에서는 덖음 처리 추출물과 찜/덖음 처리 추출물에서 차이가 없었고, 상기 추출물 모두 골다공증 예방, 개선 또는 치료가 가능하다고 판단된다.As a result of the evaluation, when MG-63 cells, which are osteoblast-like cells, were differentiated and treated with camellia young leaf extract, respectively, ALP activity was higher than that of the differentiation culture medium when the extract was treated, confirming that osteoblast differentiation was better. . However, unlike the darkest ALP staining in the steamed/roasted extract, there was no difference in ALP activity between the steamed/roasted extract and the steamed/roasted extract, and all of the extracts are considered to be capable of preventing, improving or treating osteoporosis.
<실험예 3> 동백 성엽 차의 농도에 따른 조골세포 분화 개선 효과 평가<Experimental Example 3> Evaluation of the osteoblast differentiation improvement effect according to the concentration of camellia seongyeop tea
실시예 8 내지 18의 추출물의 농도를 각각 10 μg/mL 및 20 μg/mL으로 설정하고, 이를 이용하여 조골세포 분화 개선 효능을 평가하였다. The concentrations of the extracts of Examples 8 to 18 were set to 10 μg/mL and 20 μg/mL, respectively, and the efficacy of improving osteoblast differentiation was evaluated using this.
조골세포 분화 정도를 분석하기 위해서 ALP 염색 키트인 TRACP & ALP double-stain Kit (MK300, TaKaRa)를 이용하였다. 6일 후 배양액을 제거하고 PBS로 씻어준 다음 고정액 200μl를 각 well에 넣어주고 5분간 반응시켜 고정해주었다. 2ml의 멸균수를 이용하여 2번 씻어준 다음 ALP 기질을 200μl 넣어주고 37℃에서 30분간 반응시켰다. In order to analyze the degree of osteoblast differentiation, an ALP staining kit, TRACP & ALP double-stain kit (MK300, TaKaRa) was used. After 6 days, the culture medium was removed, washed with PBS, and 200 μl of the fixative was added to each well, followed by reaction for 5 minutes to fix. After washing twice with 2ml of sterile water, 200μl of ALP substrate was added and reacted at 37℃ for 30 minutes.
분화 6일 후의 평가 결과는 도 3에 나타내었고, 염색이 진하게 될수록 ALP 활성이 높고 조골세포로 분화가 잘 된 것이다.The evaluation results after 6 days of differentiation are shown in FIG. 3, and the darker the staining, the higher the ALP activity and the better the differentiation into osteoblasts.
평가 결과, 조골 유사 세포인 MG-63세포를 분화 유도하고 동백 성엽 추출물을 각각 처리했을 때 추출물 처리시 분화 배양액에 비해 조골세포 분화가 더욱 잘 되는 것을 확인할 수 있었다. 어린잎 추출물과는 다르게 성엽 추출물의 경우 찜 처리 추출물이 다른 추출물에 비해 가장 분화를 잘 유도했다. 이러한 결과를 통해 동백 잎 추출물은 처리 방법에 관계없이 골다공증을 예방, 개선 또는 치료할 수 있는 효과가 우수하다고 판단된다. As a result of the evaluation, when MG-63 cells, which are osteoblast-like cells, were differentiated and treated with the camellia seongyeop extract, respectively, it was confirmed that the differentiation of osteoblasts was better than that of the differentiated culture medium when the extract was treated. Unlike the young leaf extract, the steamed extract induced the best differentiation compared to other extracts. Through these results, it is judged that the camellia leaf extract has an excellent effect in preventing, improving or treating osteoporosis regardless of the treatment method.
<실험예 4> 동백 어린잎의 찜/덖음 처리에 따른 조골세포 분화 개선 효과 평가<Experimental Example 4> Evaluation of osteoblast differentiation improvement effect according to steaming / roasting treatment of camellia young leaves
동백 어린잎 추출물 중 조골세포 분화 개선능이 가장 우수하였던 건신엽 찜/덖음 처리 추출물(실시예 7)을 이용하여 시간에 따른 분화 개선능을 확인하였다. Among camellia young leaf extracts, the ability to improve differentiation over time was confirmed using the steamed/roasted extract of dried leaves, which had the highest ability to improve osteoblast differentiation (Example 7).
조골세포 분화 정도를 분석하기 위해서 ALP 염색 키트인 TRACP & ALP double-stain Kit (MK300, TaKaRa)를 이용하였다. 6일 후 배양액을 제거하고 PBS로 씻어준 다음 고정액 200μl를 각 well에 넣어주고 5분간 반응시켜 고정해주었다. 2ml의 멸균수를 이용하여 2번 씻어준 다음 ALP 기질을 200μl 넣어주고 37℃에서 30분간 반응시켰다.In order to analyze the degree of osteoblast differentiation, an ALP staining kit, TRACP & ALP double-stain kit (MK300, TaKaRa) was used. After 6 days, the culture medium was removed, washed with PBS, and 200 μl of the fixative was added to each well, followed by reaction for 5 minutes to fix. After washing twice with 2ml of sterile water, 200μl of ALP substrate was added and reacted at 37℃ for 30 minutes.
염색이 진하게 될수록 ALP 활성이 높은 것이며, 분화 3일, 6일 후의 평가 결과는 도 4에 나타내었다.The darker the staining, the higher the ALP activity, and the evaluation results after 3 days and 6 days of differentiation are shown in FIG. 4 .
평가 결과, 추출물 처리시 분화 배양액에 비해 조골세포 분화가 시간에 따라 더욱 잘 되는 것을 확인할 수 있었고, 결과적으로 동백 잎 추출물을 이용하여 골다공증 예방, 개선 또는 치료가 가능하다고 판단된다.As a result of the evaluation, it was confirmed that osteoblast differentiation is better with time than in the differentiation culture medium when the extract is treated, and as a result, it is determined that osteoporosis can be prevented, improved, or treated using the camellia leaf extract.
상기 실험 결과를 종합하면, 본 발명에 따른 모든 동백 잎 추출물은 골다공증 예방 및 개선에 효과가 있을 수 있음을 증명하였고 그 중 동백 어린잎 추출물이, 어린잎 추출물 중에서는 덖음차와 찜/덖음 처리 추출물이, 특히, 이 중에서 찜/덖음 처리 추출물이 골다공증 예방, 개선 또는 치료에 보다 효과가 있을 수 있음을 증명하였다.Summarizing the above experimental results, all camellia leaf extracts according to the present invention proved to be effective in preventing and improving osteoporosis. , In particular, it was proved that steamed/roasted extracts may be more effective in preventing, improving, or treating osteoporosis.
Claims (8)
상기 동백 잎은 건조시킨 1cm 이상 18cm 미만의 어린 잎을 찜/덖음 전처리한 것이며,
덖음은 200 내지 240℃에서 1 내지 10분간 덖음하는 것이고, 찜은 90 내지 110℃에서 30 내지 60분간 찌는 것을 특징으로 하는 골다공증의 예방, 개선 또는 치료용 약학적 조성물.A pharmaceutical composition for preventing, improving or treating osteoporosis comprising a camellia leaf extract as an active ingredient,
The camellia leaves are pretreated by steaming / roasting dried young leaves of 1 cm or more and less than 18 cm,
A pharmaceutical composition for the prevention, improvement or treatment of osteoporosis, characterized by roasting at 200 to 240 ° C for 1 to 10 minutes, and steaming at 90 to 110 ° C for 30 to 60 minutes.
상기 동백 잎은 건조시킨 1cm 이상 18cm 미만의 어린 잎을 찜/덖음 전처리한 것이며,
덖음은 200 내지 240℃에서 1 내지 10분간 덖음하는 것이고, 찜은 90 내지 110℃에서 30 내지 60분간 찌는 것을 특징으로 하는 골다공증의 예방 또는 개선용 식품 조성물.A food composition for preventing or improving osteoporosis comprising a camellia leaf extract as an active ingredient,
The camellia leaves are pretreated by steaming / roasting dried young leaves of 1 cm or more and less than 18 cm,
Roasting is to roast at 200 to 240 ° C for 1 to 10 minutes, and steaming is a food composition for preventing or improving osteoporosis, characterized in that steaming at 90 to 110 ° C for 30 to 60 minutes.
(a) 동백 잎 준비 단계;
(b) 건조된 동백 잎을 찜/덖음 처리로 전처리하는 단계;
(c) 전처리된 동백 잎을 추출하는 단계;
상기 동백 잎은 건조시킨 1cm 이상 18cm 미만의 어린 잎이며,
덖음은 200 내지 240℃에서 1 내지 10분간 덖음하는 것이고, 찜은 90 내지 110℃에서 30 내지 60분간 찌는 것을 특징으로 하는 동백 잎 추출물의 제조방법.A method for producing the camellia leaf extract of claim 1 or 2, comprising the following steps:
(a) Camellia leaf preparation step;
(b) pre-treating the dried camellia leaves by steaming/roasting;
(c) extracting the pretreated camellia leaves;
The camellia leaves are dried young leaves of 1 cm or more and less than 18 cm,
The method of producing a camellia leaf extract, characterized in that the roasting is roasting at 200 to 240 ° C for 1 to 10 minutes, and the steaming is steaming at 90 to 110 ° C for 30 to 60 minutes.
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KR100702327B1 (en) | 2005-07-22 | 2007-03-30 | 보령메디앙스 주식회사 | Cosmetic material including extract of camellia flower for skin abirritation |
KR101838354B1 (en) | 2015-09-18 | 2018-03-15 | 주식회사 바이오에프디엔씨 | Anti-aging composition for skin external application comprising Camellia japonica Plant Cell Culture Extract and Method for Preparing the Same |
JP2018080166A (en) * | 2016-11-07 | 2018-05-24 | 石川県公立大学法人 | Bone metabolism regulator |
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KR100702327B1 (en) | 2005-07-22 | 2007-03-30 | 보령메디앙스 주식회사 | Cosmetic material including extract of camellia flower for skin abirritation |
KR101838354B1 (en) | 2015-09-18 | 2018-03-15 | 주식회사 바이오에프디엔씨 | Anti-aging composition for skin external application comprising Camellia japonica Plant Cell Culture Extract and Method for Preparing the Same |
JP2018080166A (en) * | 2016-11-07 | 2018-05-24 | 石川県公立大学法人 | Bone metabolism regulator |
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