KR102448913B1 - Composition comprising extract of Abeliophyllum distichum for prevention or treatment of bone diseases - Google Patents
Composition comprising extract of Abeliophyllum distichum for prevention or treatment of bone diseases Download PDFInfo
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- KR102448913B1 KR102448913B1 KR1020200058295A KR20200058295A KR102448913B1 KR 102448913 B1 KR102448913 B1 KR 102448913B1 KR 1020200058295 A KR1020200058295 A KR 1020200058295A KR 20200058295 A KR20200058295 A KR 20200058295A KR 102448913 B1 KR102448913 B1 KR 102448913B1
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- bone
- extract
- present
- bone disease
- disease
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Abstract
본 발명은 미선나무 추출물을 유효성분으로 포함하는 골질환의 예방 또는 치료용 조성물에 관한 것으로, 구체적으로 본 발명은 미선나무 추출물을 유효성분으로 포함하는 골질환의 예방 또는 치료용 약학적 조성물 및 건강기능식품에 관한 것이다. 본 발명에 따른 미선나무 추출물은 파골세포의 분화 또는 형성을 억제하는 활성이 우수하고 세포독성이 없으며 부작용을 유발시키지 않아 체내 안정성이 있으므로, 골대사 이상으로 유발되는 각종 골질환에 대한 예방, 치료 및 개선을 목적으로 하는 의약품 및 건강기능식품의 제조에 유용하게 사용할 수 있다. The present invention relates to a composition for the prevention or treatment of bone disease comprising a Miseon tree extract as an active ingredient, and more specifically, the present invention relates to a pharmaceutical composition for preventing or treating bone disease comprising the Miseon tree extract as an active ingredient, and health It is about functional food. The Miseon tree extract according to the present invention has excellent activity for inhibiting the differentiation or formation of osteoclasts, has no cytotoxicity, and has stability in the body by not causing side effects. It can be usefully used in the manufacture of pharmaceuticals and health functional foods for the purpose of
Description
본 발명은 미선나무 추출물을 유효성분으로 포함하는 골질환의 예방 또는 치료용 조성물에 관한 것이다.The present invention relates to a composition for the prevention or treatment of bone diseases comprising the extract of Miseon tree as an active ingredient.
골(bone)은 근육과 연합되어 있는 석회화된 결체조직으로서 표면은 두껍고 단단한 석회화 조직으로 신체균형을 이루는 지주역할을 하며 장기를 보호한다. 골의 안쪽은 골수조직으로 칼슘대사의 중심이 되는 부위이다. 골은 콜라겐(collagen), 오스테오칼신(osteocalcin), 오스테오넥틴(osteonectin) 등 유기질(organic substance)과 칼슘, 인, 불소 등의 무기질(inorganic substance) 및 수분으로 구성되어 있다. 생체에서는 항상 골형성(bone formation)과 골흡수(bone resorption)가 일어나고 있다. 골형성은 소량의 부갑상선 호르몬이나 안드로겐, 에스트로겐, 불소, 인 등에 의하여 촉진되며, 골흡수는 물리적 감압, 무중력상태, 다량의 부갑상선 호르몬, 부신피질 스테로이드 등에 의하여 촉진된다. 따라서 골대사에는 이들 사이의 균형(balance)이 중요하다. 그러므로 골의 항상성은 파골세포(osteoclast)에 의한 골 흡수(bone resoprtion)와 조골세포(osteoblast)에 의한 골 형성(bone formation)의 대등한 작용에 의한 리모델링 과정(bone remodeling)이 지속적으로 조절되어 유지된다.Bone is a calcified connective tissue associated with muscle, and the surface is a thick and hard calcified tissue that serves as a support for body balance and protects organs. The inside of the bone is the bone marrow tissue and is the center of calcium metabolism. Bone is composed of an organic substance such as collagen, osteocalcin, and osteonectin, an inorganic substance such as calcium, phosphorus, and fluorine, and water. In the living body, bone formation and bone resorption are always taking place. Osteogenesis is promoted by a small amount of parathyroid hormone, androgens, estrogen, fluorine, phosphorus, etc., and bone resorption is promoted by physical decompression, weightlessness, a large amount of parathyroid hormone and adrenocortical steroids. Therefore, the balance between them is important for bone metabolism. Therefore, bone homeostasis is maintained by continuously regulating the remodeling process (bone remodeling) by the equal action of bone resorption by osteoclasts and bone formation by osteoblasts. do.
이러한 뼈의 재형성, 즉 리모델링 과정에는 크게 두 종류의 세포가 관여하는 것으로 알려져 있다. 두 세포 중 하나는 뼈를 생성하는 조골세포(osteoblast)이고, 다른 하나는 뼈를 파괴하는 파골세포(osteoclast)이다. 조골세포는 RANKL(receptor activator of NF-κB ligand)과 이것의 유도 수용체(decoy receptor)인 OPG(osteoprotegerin)을 생성한다. RANKL이 파골 전구세포(osteoclast progenitor cells) 표면에 있는 수용체인 RANK(receptor activator of NF-κB)에 결합하면 파골 전구세포가 파골세포로 분화되어 골 흡수(resorption)가 일어난다. 자세하게는 RANKL 자극에 의한 세포 내 칼슘 신호 전달로 파골 전구세포에서 파골세포의 분화를 조절하는 핵심 전사인자로 알려져 있는 NFAT(nuclear factor of activated T cells)의 발현이 유도되고, 그 후 세포 외 신호조절인산화효소(extracellular signal-regulated kinase, ERK), NF-κB, p38 MAP kinase 등의 활성화가 NFAT의 자가증폭(autoamplification)을 유발하여 활성이 지속되는 것이다. 따라서 오래된 뼈의 흡수 또는 파괴는 상기와 같은 기작을 통해 파골세포에 의해 이루어지며, 이는 뼈에 구멍을 내어 적은 양의 칼슘이 혈류로 방출되어 신체기능을 유지하는데 사용된다.It is known that two types of cells are largely involved in this bone remodeling, that is, the remodeling process. One of the two cells is an osteoblast that creates bone, and the other is an osteoclast that destroys the bone. Osteoblasts produce RANKL (receptor activator of NF-κB ligand) and its decoy receptor OPG (osteoprotegerin). When RANKL binds to receptor activator of NF-κB (RANK), a receptor on the surface of osteoclast progenitor cells, osteoclasts are differentiated into osteoclasts and bone resorption occurs. Specifically, intracellular calcium signal transduction by RANKL stimulation induces the expression of NFAT (nuclear factor of activated T cells), which is known as a key transcription factor regulating the differentiation of osteoclasts from osteoclasts, and then regulates extracellular signals. Activation of extracellular signal-regulated kinase (ERK), NF-κB, p38 MAP kinase, etc. induces autoamplification of NFAT, and thus the activity is continued. Therefore, resorption or destruction of old bone is made by osteoclasts through the above mechanism, which makes a hole in the bone and a small amount of calcium is released into the bloodstream, which is used to maintain body functions.
또한 조골세포는 교원질로 구멍을 채우고 칼슘과 인의 침적물(hydroxyapatite)을 덮어서 단단한 새로운 뼈를 만들어 골격을 재건하게 된다. 성인의 경우 매년 골격의 약 10-30%가 이런 과정을 통하여 재형성되며, 파골 속도와 조골 속도가 동일해야만 전과 같은 골밀도를 유지할 수 있다. 정상 성인에서는 골 흡수 양과 골 형성 양 사이에는 항상 균형이 유지되고 있다. 이와 같이 중요한 뼈에 균형이 깨질 경우 많은 질병이 야기될 수 있는데, 이러한 질환을 골 질환이라 한다.In addition, osteoblasts fill the pores with collagen and cover with calcium and phosphorus deposits (hydroxyapatite) to form new hard bones and rebuild the skeleton. In the case of adults, about 10-30% of the skeleton is remodeled every year through this process, and the bone density as before can be maintained only when the osteoclast rate and the osteoblast rate are the same. In normal adults, there is always a balance between the amount of bone resorption and the amount of bone formation. When the balance of these important bones is broken, many diseases can be caused, and these diseases are called bone diseases.
한편, 최근에는 골질환 치료제가 갖는 부작용을 최소하기 위하여 독성이 적고, 일상생활에서 식용으로 섭취 가능한 식품과 약용식물 등 천연물 소재를 이용하여 골 손실을 최소화하고 더불어 골 형성을 촉진시킬 수 있는 새로운 골질환 예방 또는 치료 물질에 대한 연구가 다양하게 시도되고 있는 실정이나, 지금까지 미선나무를 대상으로 골질환 치료제로서의 사용 가능성에 대한 연구가 시도된 바가 없다.On the other hand, recently, in order to minimize the side effects of bone disease treatments, it is less toxic and uses natural materials such as foods and medicinal plants that can be eaten in daily life to minimize bone loss and promote bone formation. Although research on disease prevention or treatment substances is being attempted in various ways, there has been no attempt to study the possibility of using the bald eagle as a therapeutic agent for bone diseases so far.
이에 본 발명자들은 미선나무의 추출물을 제조하고 이들에 대한 파골세포 분화 억제 활성여부를 분석한 결과, 미선나무 추출물이 우수한 파골세포 분화 억제가 있음을 확인함으로써 골질환의 새로운 치료제로서의 사용 가능성을 규명함에 따라 본 발명을 완성하였다.Accordingly, the present inventors prepared extracts of Miseon tree and analyzed whether they inhibit osteoclast differentiation activity, and as a result, confirmed that Miseon tree extract has excellent inhibition of osteoclast differentiation, thereby identifying the possibility of use as a new therapeutic agent for bone diseases. Accordingly, the present invention was completed.
따라서 본 발명의 목적은 미선나무 추출물을 유효성분으로 포함하는, 골질환의 예방 또는 치료용 약학적 조성물을 제공하는 것이다.Accordingly, it is an object of the present invention to provide a pharmaceutical composition for the prevention or treatment of bone diseases, comprising the extract of Miseon tree as an active ingredient.
본 발명의 다른 목적은 미선나무 추출물을 유효성분으로 포함하는, 골질환의 예방 또는 개선용 건강기능식품을 제공하는 것이다.Another object of the present invention is to provide a health functional food for the prevention or improvement of bone disease, comprising the extract of Miseon tree as an active ingredient.
상기와 같은 본 발명의 목적을 달성하기 위해서, 본 발명은 미선나무 추출물을 유효성분으로 포함하는, 골질환의 예방 또는 치료용 약학적 조성물을 제공한다.In order to achieve the object of the present invention as described above, the present invention provides a pharmaceutical composition for the prevention or treatment of bone diseases, comprising an extract of Miseon tree as an active ingredient.
본 발명의 일실시예에 있어서, 상기 추출물은 미선나무의 잎 추출물일 수 있다.In one embodiment of the present invention, the extract may be a leaf extract of Miseon tree.
본 발명의 일실시예에 있어서, 상기 추출물은 물, 메탄올, 에탄올, 프로판올, 이소프로판올, 부탄올, 아세톤, 에테르, 벤젠, 클로로포름, 에틸아세테이트, 메틸렌클로라이드, 헥산 및 시클로헥산으로 이루어진 군 중에서 선택되는 용매를 이용하여 수득한 것일 수 있다.In one embodiment of the present invention, the extract is a solvent selected from the group consisting of water, methanol, ethanol, propanol, isopropanol, butanol, acetone, ether, benzene, chloroform, ethyl acetate, methylene chloride, hexane and cyclohexane It may be obtained using
본 발명의 일실시예에 있어서, 상기 추출물은 파골세포의 분화 또는 형성을 억제하는 것일 수 있다.In one embodiment of the present invention, the extract may be to inhibit the differentiation or formation of osteoclasts.
본 발명의 일실시예에 있어서, 상기 골질환은 골다공증, 파세트 골질환, 구루병, 골연화증, 신부전 환자의 신성골이영양증, 류머티스성 골질환, 퇴행성 골질환, 뼈전이성 병소(bone metastatic lesion), 원발성으로 뼈에 생성된 종양, 치주질환, 염증성 치조골 흡수질환 및 염증성 뼈 흡수질환으로 이루어진 군 중에서 선택되는 것일 수 있다.In one embodiment of the present invention, the bone disease is osteoporosis, facet bone disease, rickets, osteomalacia, renal osteodystrophy in patients with renal failure, rheumatic bone disease, degenerative bone disease, bone metastatic lesion, primary As a result, it may be selected from the group consisting of a bone-generated tumor, periodontal disease, inflammatory alveolar bone resorption disease, and inflammatory bone resorption disease.
또한 본 발명은 미선나무 추출물을 유효성분으로 포함하는, 골질환의 예방 또는 개선용 건강기능식품을 제공한다.In addition, the present invention provides a health functional food for the prevention or improvement of bone diseases, comprising the extract of Miseon tree as an active ingredient.
본 발명의 일실시예에 있어서, 상기 추출물은 미선나무의 잎 추출물일 수 있다.In one embodiment of the present invention, the extract may be a leaf extract of Miseon tree.
본 발명의 일실시예에 있어서, 상기 추출물은 물, 메탄올, 에탄올, 프로판올, 이소프로판올, 부탄올, 아세톤, 에테르, 벤젠, 클로로포름, 에틸아세테이트, 메틸렌클로라이드, 헥산 및 시클로헥산으로 이루어진 군 중에서 선택되는 용매를 이용하여 수득한 것일 수 있다. In one embodiment of the present invention, the extract is a solvent selected from the group consisting of water, methanol, ethanol, propanol, isopropanol, butanol, acetone, ether, benzene, chloroform, ethyl acetate, methylene chloride, hexane and cyclohexane It may be obtained using
본 발명의 일실시예에 있어서, 상기 추출물은 파골세포의 분화 또는 형성을 억제하는 것일 수 있다.In one embodiment of the present invention, the extract may be to inhibit the differentiation or formation of osteoclasts.
본 발명의 일실시예에 있어서, 상기 골질환은 골다공증, 파세트 골질환, 구루병, 골연화증, 신부전 환자의 신성골이영양증, 류머티스성 골질환, 퇴행성 골질환, 뼈전이성 병소(bone metastatic lesion), 원발성으로 뼈에 생성된 종양, 치주질환, 염증성 치조골 흡수질환 및 염증성 뼈 흡수질환으로 이루어진 군 중에서 선택되는 것일 수 있다.In one embodiment of the present invention, the bone disease is osteoporosis, facet bone disease, rickets, osteomalacia, renal osteodystrophy in patients with renal failure, rheumatic bone disease, degenerative bone disease, bone metastatic lesion, primary As a result, it may be selected from the group consisting of a bone-generated tumor, periodontal disease, inflammatory alveolar bone resorption disease, and inflammatory bone resorption disease.
본 발명에 따른 미선나무 추출물은 파골세포의 분화 또는 형성을 억제하는 활성이 우수하고 세포독성이 없으며 부작용을 유발시키지 않아 체내 안정성이 있으므로, 골대사 이상으로 유발되는 각종 골질환에 대한 예방, 치료 및 개선을 목적으로 하는 의약품 및 건강기능식품의 제조에 유용하게 사용할 수 있다. The Miseon tree extract according to the present invention has excellent activity for inhibiting the differentiation or formation of osteoclasts, has no cytotoxicity, and has stability in the body by not causing side effects. It can be usefully used in the manufacture of pharmaceuticals and health functional foods for the purpose of
도 1은 본 발명의 일실시예에서 미선나무의 열매, 줄기 및 잎에 대한 각 부위를 이용하여 추출물을 수득하는 과정을 모식도로 나타낸 것이다.
도 2는 본 발명의 일실시예에서 미선나무의 각 부위별 메탄올 추출물에 대한 파골세포 분화 억제 활성 및 세포독성 여부를 분석한 결과를 나타낸 것이다.
도 3은 본 발명의 일실시예에서 용매 종류별 미선나무 잎 추출물에 대한 파골세포 분화 억제 활성을 나타낸 것이다.1 is a schematic diagram showing the process of obtaining an extract using each part for the fruit, stem and leaf of Miseon tree in an embodiment of the present invention.
Figure 2 shows the results of analyzing the osteoclast differentiation inhibitory activity and cytotoxicity of the methanol extract for each part of Miseon tree in an embodiment of the present invention.
Figure 3 shows the osteoclast differentiation inhibitory activity on the leaf extract of Miseon tree by type of solvent in one embodiment of the present invention.
본 발명은 파골세포의 형성을 효과적으로 억제하여 골질환을 예방 또는 치료할 수 있는 미선나무 추출물의 새로운 용도를 규명한 점에 특징이 있다.The present invention is characterized in that it has identified a new use of the Miseon tree extract, which can prevent or treat bone diseases by effectively inhibiting the formation of osteoclasts.
상기 미선나무(Abeliophyllum distichum)는 우리나라에서 자생하는 식물로서, 항암 활성, 아토피 치료 효능, 탈모방지와 같은 효능이 알려져 있으나 아직까지 많은 연구가 진행되고 있지 못하다. The Abeliophyllum distichum is a plant that grows in Korea and is known for its anti-cancer activity, atopy treatment efficacy, and anti-hair loss, but many studies have not been conducted yet.
따라서 우리의 자원인 미선나무에 대한 다양한 효능을 밝히고, 이를 적극적으로 번식시켜 활용하기 위한 연구가 필요하다.Therefore, there is a need for research to reveal various effects of Miseon tree, our resource, and to actively propagate and utilize it.
이에 본 발명자들은 이러한 미선나무의 유용한 효능을 연구하던 중, 미선나무의 추출물이 파골세포의 분화를 효과적으로 억제하는 활성이 있음을 최초로 규명함으로써, 미선나무 추출물을 유효성분으로 포함하는 골질환의 예방 또는 치료 용도로 사용할 수 있음을 확인하였다. Accordingly, the present inventors first identified that the extract of Miseon tree effectively inhibits the differentiation of osteoclasts while studying the useful efficacy of the Miseon tree, thereby preventing or preventing bone disease containing Miseon tree extract as an active ingredient. It was confirmed that it can be used for therapeutic purposes.
본 발명에서 상기 “파골세포”란, 골 기질을 파괴하고 뼈의 미네랄을 분해하는 역할을 담당하여 조직의 파괴 및 흡수 기능을 수행하는 세포를 의미한다. 상기 파골세포는 골 내에서 조골세포와의 불균형이 발생하는 경우 골질환을 유발할 수 있는 것으로 보고되어 있다.In the present invention, the “osteoclast” refers to cells that destroy the bone matrix and play a role in decomposing bone minerals to perform tissue destruction and absorption functions. It has been reported that the osteoclasts can cause bone disease when an imbalance with osteoblasts occurs in the bone.
상기 “조골세포”는 척추동물들의 골세포를 만드는 세포로, 간엽줄기세포에서 기원하여 형성되는데 조골세포의 분화에 의한 칼슘형성과 같은 무기질화는 뼈의 강도를 유지시켜줄 뿐만 아니라, 신체 전체의 칼슘 및 호르몬 대사의 항상성에도 매우 중요한 기능을 한다. 조골세포의 분화에 의한 칼슘형성은 비타민 D 및 부갑상선 호르몬(parathyroid hormone) 등에 의해 조절되며, 조골세포의 분화에 의한 골형성은 세포 내에서 뼈 형태형성 단백질(bone morphogenetic protein; BMP), Wnt, MAP 키나아제, 칼시뉴린-칼모듈린 키나아제(calcineurin-calmodulin kinase), NF-κB, AP-1 등의 다양한 신호전달 체계의 상호작용(cross-talk)에 의해 조골세포의 분화와 관련된 알칼린 포스파타제(alkaline phosphatase, ALP)가 초기 분화단계에서 합성된 후, 무기질화와 관련된 오스테오폰틴(osteopontin), 오스테오칼신(osteocalcin), 제1형 콜라겐(type 1 collagen) 등이 합성됨으로써 이루어진다고 알려져 있다. 따라서 이러한 조골세포의 분화와 형성에 이상이 초래하게 되면 뼈 형성에 문제가 발생하여 골질환이 유발된다.The “osteoblast” is a cell that makes osteocytes of vertebrates, and is formed by originating from mesenchymal stem cells. Mineralization such as calcium formation by the differentiation of osteoblasts not only maintains bone strength, but also provides calcium and It also plays a very important role in the homeostasis of hormonal metabolism. Calcium formation by the differentiation of osteoblasts is regulated by vitamin D and parathyroid hormone, and the formation of bone by differentiation of osteoblasts within the cells is bone morphogenetic protein (BMP), Wnt, MAP. Alkaline phosphatase related to the differentiation of osteoblasts by the interaction (cross-talk) of various signaling systems such as kinase, calcineurin-calmodulin kinase, NF-κB, and AP-1. It is known that phosphatase, ALP) is synthesized in the initial differentiation stage, and then, osteopontin, osteocalcin,
이와 같이 상기 조골세포와 파골세포의 상호작용의 균형이 깨지게 되면 전신적(systemic) 또는 국부적(local) 골질환을 일으키게 되며, 이러한 질환들은 대부분 파골세포의 수와 활성의 증가로 인한 것으로 알려져 있다. As such, when the balance of the interaction between the osteoclasts and osteoclasts is broken, systemic or local bone diseases are caused, and these diseases are mostly known to be due to an increase in the number and activity of osteoclasts.
따라서 이러한 파골세포의 형성 및 분화를 억제한다면 파골세포로 인해 유발되는 골질환을 예방 또는 치료할 수 있다.Therefore, if the formation and differentiation of these osteoclasts is suppressed, bone diseases caused by osteoclasts can be prevented or treated.
또한 본 발명의 다른 일실시예에서는, 미선나무 추출물의 파골세포 분화억제 활성에 대해, 미선나무의 부위별 추출물에 대한 효능을 확인한 결과, 미선나무의 잎이 줄기 및 열매 추출물에 비해 더 우수한 파골세포 분화 억제 활성이 있는 것으로 나타났다.Also, in another embodiment of the present invention, for the osteoclast differentiation inhibitory activity of the Miseon tree extract, as a result of confirming the efficacy of the extract for each part of Miseon tree, the leaf of Miseon tree is superior to osteoclasts compared to the stem and fruit extracts. It was shown to have differentiation inhibitory activity.
그러므로 본 발명은 미선나무 추출물을 유효성분으로 포함하는, 골질환의 예방 또는 치료용 약학적 조성물을 제공한다.Therefore, the present invention provides a pharmaceutical composition for the prevention or treatment of bone diseases, comprising the extract of Miseon tree as an active ingredient.
본 발명에서 미선나무 추출물로 예방, 개선 또는 치료하고자 하는 상기 골질환은 조골세포와 파골세포의 상호작용의 불균형으로 유발될 수 있는 질환 및 과도한 파골세포의 분화 및 활성으로 유발될 수 있는 질환이라면 모두 포함할 수 있으며, 이에 제한되지는 않으나, 골다공증, 파세트 골질환, 구루병, 골연화증, 신부전 환자의 신성골이영양증, 류머티스성 골질환, 퇴행성 골질환, 뼈전이성 병소(bone metastatic lesion), 원발성으로 뼈에 생성된 종양, 치주질환, 염증성 치조골 흡수질환 및 염증성 뼈 흡수질환을 포함할 수 있다. In the present invention, the bone disease to be prevented, improved or treated with the extract of Miseon tree is any disease that can be caused by an imbalance in the interaction between osteoblasts and osteoclasts and a disease that can be caused by excessive differentiation and activity of osteoclasts. It may include, but is not limited to, osteoporosis, facet bone disease, rickets, osteomalacia, renal osteodystrophy in patients with renal failure, rheumatic bone disease, degenerative bone disease, bone metastatic lesion, primary bone It may include tumors generated in the cyst, periodontal disease, inflammatory alveolar bone resorption disease, and inflammatory bone resorption disease.
또한 본 발명에 따른 상기 미선나무 추출물은 당업계에 공지된 추출 및 분리하는 방법을 사용하여 천연으로부터 추출 및 분리하여 수득한 것을 사용할 수 있으며, 본 발명에서 정의된 추출물은 적절한 용매를 이용하여 미선나무로부터 추출한 것이며, 예를 들어, 미선나무의 조추출물, 극성용매 가용 추출물 또는 비극성용매 가용 추출물을 모두 포함한다. In addition, the extract of Miseon tree according to the present invention may be obtained by extraction and separation from nature using extraction and separation methods known in the art, and the extract defined in the present invention may be obtained by using an appropriate solvent. It is extracted from, for example, includes all of the crude extract of Miseon tree, polar solvent-soluble extract or non-polar solvent-soluble extract.
상기 미선나무로부터 추출물을 수득하기 위한 적절한 용매로는 약학적으로 허용되는 유기용매라면 어느 것을 사용해도 무방하며, 물 또는 유기용매를 사용할 수 있으며, 이에 제한되지는 않으나, 예를 들어, 정제수, 메탄올(methanol), 에탄올(ethanol), 프로판올(propanol), 이소프로판올(isopropanol), 부탄올(butanol) 등을 포함하는 탄소수 1 내지 4의 알코올, 아세톤(acetone), 에테르(ether), 벤젠(benzene), 클로로포름(chloroform), 에틸아세테이트(ethyl acetate), 메틸렌클로라이드(methylene chloride), 헥산(hexane) 및 시클로헥산(cyclohexane) 등의 각종 용매를 단독으로 혹은 혼합하여 사용할 수 있다. 바람직하게는 메탄올을 사용할 수 있다.As a suitable solvent for obtaining the extract from the Miseon tree, any pharmaceutically acceptable organic solvent may be used, and water or an organic solvent may be used, but is not limited thereto, for example, purified water, methanol (methanol), ethanol (ethanol), propanol (propanol), isopropanol (isopropanol), alcohol having 1 to 4 carbon atoms including butanol (butanol), acetone (acetone), ether (ether), benzene (benzene), chloroform Various solvents such as (chloroform), ethyl acetate, methylene chloride, hexane and cyclohexane may be used alone or in combination. Preferably, methanol can be used.
추출 방법으로는 열수추출법, 냉침추출법, 환류냉각추출법, 용매추출법, 수증기증류법, 초음파추출법, 용출법, 압착법 등의 방법 중 어느 하나를 선택하여 사용할 수 있다. 또한, 목적하는 추출물은 추가로 통상의 분획 공정을 수행할 수도 있으며, 통상의 정제 방법을 이용하여 정제될 수도 있다. As the extraction method, any one of methods such as hot water extraction, cold extraction, reflux cooling extraction, solvent extraction, steam distillation, ultrasonic extraction, elution, and compression may be selected and used. In addition, the desired extract may be further subjected to a conventional fractionation process, and may be purified using a conventional purification method.
본 발명의 다른 일실시예에서는, 미선나무 잎 추출물에 대한 파골세포 분화 억제 효능과 관련하여 가장 우수한 파골세포 분화 억제 활성을 갖는 추출물을 수득하기 위해 다양한 용매를 이용하여 추출물을 수득하고 그에 대한 효능 분석을 수행하였는데, 다른 종류의 용매에 비해 메탄올을 사용한 경우 가장 우수한 파골세포 분화 억제 활성이 있는 것으로 나타났다.In another embodiment of the present invention, in order to obtain an extract having the best osteoclast differentiation inhibitory activity with respect to the osteoclast differentiation inhibitory effect on the Miseon tree leaf extract, the extract is obtained using various solvents and the efficacy analysis thereof was performed, and it was found that methanol had the most excellent osteoclast differentiation inhibitory activity compared to other types of solvents.
따라서 이러한 결과를 통해, 본 발명자들은 미선나무의 잎을 대상으로 메탄올을 첨가하여 수득한 추출물, 즉 상기 추출물에 함유된 미선나무 잎 유래의 유용성분들이 파골세포의 분화를 효과적으로 억제할 수 있으므로, 골질환의 새로운 치료제로서 가장 유용하게 사용할 수 있음을 알 수 있었다. Therefore, through these results, the present inventors have found that the extract obtained by adding methanol to the leaves of Miseon tree, that is, useful components derived from Miseon wood leaves contained in the extract can effectively inhibit the differentiation of osteoclasts, It was found that it can be most usefully used as a new therapeutic agent for diseases.
본 발명의 상기 약학적 조성물은 본 발명의 미선나무 추출물을 유효성분으로 포함하는 약제학적 조성물로서 이러한 본 발명의 유효성분 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.The pharmaceutical composition of the present invention is a pharmaceutical composition comprising the Miseon tree extract of the present invention as an active ingredient. As the auxiliary agent, an excipient, a disintegrant, a sweetener, a binder, a coating agent, an expanding agent, a lubricant, a lubricant, or a flavoring agent may be used.
상기 약제학적 조성물은 투여를 위해서 상기 본 발명의 유효성분 이외에 추가로 약제학적으로 허용 가능한 담체를 1종 이상 포함하여 약제학적 조성물로 바람직하게 제제화할 수 있다.The pharmaceutical composition may be preferably formulated into a pharmaceutical composition including one or more pharmaceutically acceptable carriers in addition to the active ingredient of the present invention 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. 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. 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 포함한다. 붕해제는 이에 제한되는 것은 아니나, 녹말, 메틸 셀룰로스, 아가, 벤토니트, 잔탄 검 등을 포함한다. 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. 또한 희석제, 분산제, 계면활성제, 결합제 및 윤활제를 부가적으로 첨가하여 수용액, 현탁액, 유탁액 등과 같은 주사용 제형, 환약, 캡슐, 과립 또는 정제로 제제화할 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.Formulations of the pharmaceutical composition may be granules, powders, tablets, coated tablets, capsules, suppositories, solutions, syrups, juices, suspensions, emulsions, drops or injectable solutions. For formulation in the form of, for example, tablets or capsules, the active ingredient may be combined with an orally, non-toxic, pharmaceutically acceptable inert carrier such as ethanol, glycerol, water, and the like. In addition, if desired or required, suitable binders, lubricants, disintegrants and color-developers may also be included in the mixture. Suitable binders include, but are not limited to, starch, gelatin, natural sugars such as glucose or beta-lactose, corn sweeteners, natural and synthetic gums such as acacia, tracacanth or sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride, and the like. Disintegrants include, but are not limited to, starch, methyl cellulose, agar, bentonite, xanthan gum, and the like. In the composition formulated as a liquid solution, acceptable pharmaceutical carriers are sterile and biocompatible, and include saline, sterile water, Ringer's solution, buffered saline, albumin injection, dextrose solution, maltodextrin solution, glycerol, ethanol and One or more of these components may be mixed and used, and other conventional additives such as antioxidants, buffers, and bacteriostats may be added as needed. In addition, diluents, dispersants, surfactants, binders and lubricants may be additionally added to form an injectable formulation such as an aqueous solution, suspension, emulsion, etc., pills, capsules, granules or tablets. Furthermore, it can be preferably formulated according to each disease or component using the method disclosed in Remington's Pharmaceutical Science, Mack Publishing Company, Easton PA by an appropriate method in the field.
본 발명의 미선나무 추출물은 본 발명의 조성물에 1 내지 100㎍/㎖의 농도로 포함될 수 있으며, 또한 본 발명의 미선나무 추출물은 조성물 총 중량에 대하여 0.1~95 중량%로 포함될 수 있다.The Miseon tree extract of the present invention may be included in the composition of the present invention at a concentration of 1 to 100 μg/ml, and the Miseon tree extract of the present invention may be included in an amount of 0.1 to 95% by weight based on the total weight of the composition.
또한 본 발명의 조성물은 식품 조성물일 수 있는데, 식품 조성물은 유효성분인 미선나무 추출물을 함유하는 것 외에 통상의 식품 조성물과 같이 여러 가지 향미제 또는 천연 탄수화물 등을 추가 성분으로서 함유할 수 있다.In addition, the composition of the present invention may be a food composition, and the food composition may contain various flavoring agents or natural carbohydrates as an additional component like a conventional food composition in addition to containing the Miseon tree extract as an active ingredient.
상술한 천연 탄수화물의 예는 모노사카라이드, 예를 들어, 포도당, 과당 등; 디사카라이드, 예를 들어 말토스, 슈크로스 등; 및 폴리사카라이드, 예를 들어 덱스트린, 시클로덱스트린 등과 같은 통상적인 당, 및 자일리톨, 소르비톨, 에리트리톨 등의 당알콜이다. 상술한 향미제는 천연 향미제 (타우마틴), 스테비아 추출물(예를 들어 레바우디오시드 A, 글리시르히진 등) 및 합성 향미제 (사카린, 아스파르탐 등)를 유리하게 사용할 수 있다.Examples of the above-mentioned natural carbohydrates include monosaccharides such as glucose, fructose and the like; disaccharides such as maltose, sucrose and the like; and polysaccharides such as conventional sugars such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. The above-mentioned flavoring agents can advantageously use natural flavoring agents (Taumatine), stevia extract (eg rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.).
본 발명의 식품 조성물은 상기 약제학적 조성물과 동일한 방식으로 제제화되어 기능성 식품으로 이용하거나, 각종 식품에 첨가할 수 있다. 본 발명의 조성물을 첨가할 수 있는 식품으로는 예를 들어, 음료류, 육류, 초코렛, 식품류, 과자류, 피자, 라면, 기타 면류, 껌류, 사탕류, 아이스크림류, 알코올 음료류, 비타민 복합제 및 건강보조식품류 등이 있다.The food composition of the present invention may be formulated in the same manner as the 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, chocolate, foods, confectionery, pizza, ramen, other noodles, gums, candy, ice cream, alcoholic beverages, vitamin complexes and health supplements. There is this.
또한 상기 식품 조성물은 유효성분인 미선나무 추출물 외에 여러 가지 영양제, 비타민, 광물 (전해질), 합성 풍미제 및 천연 풍미제 등의 풍미제, 착색제 및 중진제 (치즈, 초콜릿 등), 펙트산 및 그의 염, 알긴산 및 그의 염, 유기산, 보호성 콜로이드 증점제, pH 조절제, 안정화제, 방부제, 글리세린, 알콜, 탄산음료에 사용되는 탄산화제 등을 함유할 수 있다. 그밖에 본 발명의 식품 조성물은 천연 과일 쥬스 및 과일 쥬스 음료 및 야채 음료의 제조를 위한 과육을 함유할 수 있다.In addition, the food composition contains various nutrients, vitamins, minerals (electrolytes), synthetic flavors and natural flavoring agents, colorants and thickeners (cheese, chocolate, etc.), pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated beverages, and the like. In addition, the food composition of the present invention may contain natural fruit juice and pulp for the production of fruit juice beverages and vegetable beverages.
이러한 본 발명의 유효성분인 미선나무 추출물은 천연소재로부터 추출된 물질로서 부작용이 거의 없으므로 장기간 복용 시에도 안심하고 사용할 수 있다. The active ingredient of the present invention, Miseon tree extract, is a material extracted from natural materials and has almost no side effects, so it can be safely used even when taken for a long period of time.
또한, 본 발명은 상기 미선나무 추출물을 유효성분으로 포함하는 골질환의 예방 또는 개선을 위한 건강기능식품을 제공한다.In addition, the present invention provides a health functional food for the prevention or improvement of bone diseases comprising the extract of Miseon tree as an active ingredient.
본 발명의 건강기능식품은 정제, 캅셀, 분말, 과립, 액상, 환 등의 형태로 제조 및 가공할 수 있다.The health functional food of the present invention can be manufactured and processed in the form of tablets, capsules, powders, granules, liquids, pills, and the like.
본 발명에서 “건강기능식품”이라 함은 건강기능식품에 관한 법률 제6727호에 따른 인체에 유용한 기능성을 가진 원료나 성분을 사용하여 제조 및 가공한 식품을 말하며, 인체의 구조 및 기능에 대하여 영양소를 조절하거나 생리학적 작용 등과 같은 보건 용도에 유용한 효과를 얻을 목적으로 섭취하는 것을 의미한다.In the present invention, the term “health functional food” refers to food manufactured and processed using raw materials or ingredients useful for the human body according to Act No. 6727 of the Health Functional Food Act, and provides nutrients for the structure and function of the human body. It refers to ingestion for the purpose of obtaining useful effects for health purposes, such as controlling or physiological effects.
본 발명의 건강기능식품은 통상의 식품 첨가물을 포함할 수 있으며, 식품 첨가물로서의 적합 여부는 다른 규정이 없는 한, 식품의약품안전청에 승인된 식품 첨가물 공전의 총칙 및 일반시험법 등에 따라 해당 품목에 관한 규격 및 기준에 의하여 판정한다.The health functional food of the present invention may contain normal food additives, and unless otherwise specified, whether it is suitable as a food additive is related to the item according to the general rules and general test method of food additives approved by the Food and Drug Administration. It is judged according to the standards and standards.
상기 “식품 첨가물 공전”에 수재된 품목으로는 예를 들어, 케톤류, 글리신, 구연산칼슘, 니코틴산, 계피산 등의 화학적 합성물; 감색소, 감초추출물, 결정셀룰로오스, 고량색소, 구아검 등의 천연첨가물; L-글루타민산나트륨제제, 면류첨가알칼리제, 보존료제제, 타르색소제제 등의 혼합제제류 등을 들 수 있다.The items listed in the “Food Additives Code” include, for example, chemical compounds such as ketones, glycine, calcium citrate, nicotinic acid, and cinnamic acid; natural additives such as persimmon pigment, licorice extract, crystalline cellulose, high pigment, and guar gum; and mixed preparations such as sodium L-glutamate preparations, noodles-added alkalis, preservatives, and tar dye preparations.
예를 들어, 정제 형태의 건강기능식품은 본 발명의 유효성분인 상기 미선나무 추출물을 부형제, 결합제, 붕해제 및 다른 첨가제와 혼합한 혼합물을 통상의 방법으로 과립화한 다음, 활택제 등을 넣어 압축성형하거나, 상기 혼합물을 직접 압축 성형할 수 있다. 또한 상기 정제 형태의 건강기능식품은 필요에 따라 교미제 등을 함유할 수도 있다.For example, a health functional food in tablet form is granulated in a conventional manner by mixing a mixture of the Miseon tree extract, which is the active ingredient of the present invention, with an excipient, binder, disintegrant and other additives, and then a lubricant is added thereto. Compression molding, or direct compression molding of the mixture. In addition, the health functional food in the form of tablets may contain a corrosive agent or the like, if necessary.
캅셀 형태의 건강기능식품 중 경질 캅셀제는 통상의 경질 캅셀에 본 발명의 유효성분인 상기 미선나무 추출물을 부형제 등의 첨가제와 혼합한 혼합물을 충진하여 제조할 수 있으며, 연질 캅셀제는 상기 미선나무 추출물을 부형제 등의 첨가제와 혼합한 혼합물을 젤라틴과 같은 캅셀기제에 충진하여 제조할 수 있다. 상기 연질 캅셀제는 필요에 따라 글리세린 또는 소르비톨 등의 가소제, 착색제, 보존제 등을 함유할 수 있다.Among health functional foods in capsule form, hard capsules can be prepared by filling a conventional hard capsule with a mixture of the Miseon tree extract, which is the active ingredient of the present invention, mixed with additives such as excipients. It can be prepared by filling a mixture mixed with additives such as excipients in a capsule base such as gelatin. The soft capsules may contain a plasticizer such as glycerin or sorbitol, a colorant, a preservative, and the like, if necessary.
환 형태의 건강기능식품은 본 발명의 유효성분과 부형제, 결합제, 붕해제 등을 혼합한 혼합물을 기존에 공지된 방법으로 성형하여 조제할 수 있으며, 필요에 따라 백당이나 다른 제피제로 제피할 수 있으며, 또는 전분, 탈크와 같은 물질로 표면을 코팅할 수도 있다.A health functional food in the form of a ring can be prepared by molding a mixture of the active ingredient of the present invention with an excipient, a binder, a disintegrant, etc. by a known method, and if necessary, it can be coated with sucrose or other peeling agent, Alternatively, the surface may be coated with a material such as starch or talc.
과립 형태의 건강기능식품은 본 발명의 유효성분과 부형제, 결합제, 붕해제 등을 혼합한 혼합물을 기존에 공지된 방법으로 입상으로 제조할 수 있으며, 필요에 따라 착향제, 교미제 등을 함유할 수 있다.The health functional food in the form of granules can be prepared in granular form by a conventionally known method by mixing the active ingredient of the present invention with an excipient, binder, disintegrant, etc. have.
본 발명의 상기 미선나무 추출물을 유효성분으로 포함하는 건강기능식품은 하기 실시예에서도 확인한 바와 같이 파골세포의 분화 및 형성 억제 활성이 우수하여, 골질환의 예방 또는 개선에 효과적이다.The health functional food containing the Miseon tree extract of the present invention as an active ingredient has excellent osteoclast differentiation and formation inhibitory activity, as confirmed in the following Examples, and is effective in preventing or improving bone diseases.
상기 건강기능식품은 음료류, 육류, 초코렛, 식품류, 과자류, 피자, 라면, 기타 면류, 껌류, 사탕류, 아이스크림류, 알코올 음료류, 비타민 복합제 및 건강보조식품류 등일 수 있다.The health functional food may be beverages, meat, chocolate, foods, confectionery, pizza, ramen, other noodles, gums, candy, ice cream, alcoholic beverages, vitamin complexes and health supplements.
이하, 실시예를 통하여 본 발명을 보다 상세히 설명하고자 한다. 이들 실시예는 본 발명을 보다 구체적으로 설명하기 위한 것으로, 본 발명의 범위가 이들 실시예에 한정되는 것은 아니다.Hereinafter, the present invention will be described in more detail through examples. These Examples are for illustrating the present invention in more detail, and the scope of the present invention is not limited to these Examples.
<실시예 1><Example 1>
미선나무 부위별 추출물의 제조Preparation of extracts for each part of Miseon tree
미선나무의 부위별 추출물을 수득하기 위해, 동결건조된 미선나무의 잎, 줄기 및 열매를(각 부위별 50g) 각각 메탄올을 이용하여 상온에서 6시간씩 3회에 걸쳐 추출하였다. 이후 각 부위별 메탄올 추출물을 Whatman 여과지를 이용하여 여과시킨 후, 남아있는 고형분은 분리하였고, 여과된 추출물을 40℃에서 감압농축기를 이용하여 감압농축하여 각 부위별 메탄올 추출물을 수득하였다. 상기 과정의 모식도는 도 1에 나타내었다.In order to obtain extracts for each part of Miseon tree, lyophilized leaves, stems and fruits (50 g for each part) of Miseon tree were each extracted three times at room temperature for 6 hours at room temperature using methanol. Thereafter, the methanol extract for each part was filtered using Whatman filter paper, the remaining solid was separated, and the filtered extract was concentrated under reduced pressure at 40° C. using a vacuum concentrator to obtain a methanol extract for each part. A schematic diagram of the above process is shown in FIG. 1 .
<실시예 2><Example 2>
미선나무의 각 부위별 추출물에 대한 파골세포 억제 효과분석Analysis of osteoclast inhibition effect on extracts of each part of Miseon tree
본 발명자들은 상기 실시예 1에서 수득한 미선나무의 각 부위별 추출물, 즉 잎, 줄기 및 열매의 메탄올 추출물에 대한 파골세포 억제 효능 여부를 분석하였다. The present inventors analyzed the osteoclast inhibitory efficacy of the extract for each part of the Miseon tree obtained in Example 1, that is, the methanol extract of leaves, stems and fruits.
이를 위해 수컷 6주령 C57BL/6 생쥐를 100% CO2 가스를 이용하여 안락사 시킨 뒤 대퇴골과 경골을 분리하였다. 분리된 경골과 대퇴골의 양끝을 절단한 뒤 1mL 주사기로 골수강 내의 세포를 채취하고 RBCL(red blood cell lysis buffer)를 이용하여 적혈구를 용해시켜 제거하였다. 이후 얻어진 골수세포를 10% FBS, M-CSF (30 ng/mL)가 포함된 α-MEM 배지에서 3일간 배양하였다. 배양 3일 후, 부착된 세포를 대식세포(bone marrow-derived macrophages, BMMs)로 실험에 사용하였다. 구체적으로 BMMs 세포를 48웰 플레이트에 분주한 후, RANKL(100ng/ml)을 첨가하여 파골세포로의 분화를 유도하였다. 이 과정에 본 발명의 미선나무 부위별 메탄올 추출물을 각 농도별(0.1, 1, 10 μg/ml)로 처리하고 3일 후에 TRAP 염색분석을 실시하였다. For this purpose, 6-week-old male C57BL/6 mice were euthanized using 100% CO 2 gas, and then the femur and tibia were separated. After cutting both ends of the separated tibia and femur, cells in the intramedullary cavity were collected with a 1mL syringe, and red blood cells were lysed and removed using RBCL (red blood cell lysis buffer). Then, the obtained bone marrow cells were cultured in α-MEM medium containing 10% FBS and M-CSF (30 ng/mL) for 3 days. After 3 days of culture, the adherent cells were used for experiments as bone marrow-derived macrophages (BMMs). Specifically, BMMs cells were seeded in a 48-well plate, and RANKL (100 ng/ml) was added to induce differentiation into osteoclasts. In this process, the methanol extract of each part of Miseon tree of the present invention was treated at each concentration (0.1, 1, 10 μg/ml), and TRAP staining analysis was performed 3 days later.
그 결과, 도 2에 나타낸 바와 같이, 미선나무의 열매 추출물은 다른부위의 추출물에 비해 파골세포 분화에 영향을 미치지 않은 것으로 나타났고, 잎과 줄기는 파골세포의 분화에 영향을 미치는 것으로 나타났다. 그러나 줄기 추출물의 경우, 세포독성이 유발되어 세포가 사멸되는 것으로 나타났다.As a result, as shown in FIG. 2 , the fruit extract of Miseon tree did not appear to have an effect on osteoclast differentiation compared to extracts from other parts, and the leaves and stems were found to affect the differentiation of osteoclasts. However, in the case of the stem extract, it was found that cytotoxicity was induced and cells were killed.
한편, 미선나무의 잎 추출물만이 세포독성의 유발 없이 파골세포를 효과적으로 억제할 수 있는 것으로 나타났다.On the other hand, it was found that only the leaf extract of Miseon tree can effectively inhibit osteoclasts without inducing cytotoxicity.
따라서 이러한 결과를 통해 본 발명자들은 미선나무 열매의 경우 파골세포 분화 억제 활성이 없으며, 줄기의 경우 세포독성이 유발되어 유해성의 문제가 발생할 수 있어 열매와 줄기 추출물은 골질환 치료제의 소재로 사용하기에는 문제가 있음을 확인하였다.Therefore, through these results, the present inventors found that, in the case of Miseon tree fruit, there is no osteoclast differentiation inhibitory activity, and in the case of stem, cytotoxicity is induced, which may cause a problem of toxicity. It was confirmed that there is
반면, 미선나무의 잎 추출물은 세포독성 없이 파골세포의 분화를 효과적으로 억제할 수 있음을 확인함으로써, 미선나무의 잎 추출물만이 안전하게 골질환의 새로운 치료제의 소재로 사용 가능함을 알 수 있었다. On the other hand, by confirming that the leaf extract of Miseon tree can effectively inhibit the differentiation of osteoclasts without cytotoxicity, it was found that only the leaf extract of Miseon tree can be safely used as a material for a new therapeutic agent for bone diseases.
<실시예 3><Example 3>
미선나무의 잎의 용매별 추출물에 대한 파골세포 억제 효과분석Analysis of osteoclast inhibition effect on extracts of leaves of Miseon tree by solvent
상기 실시예 2의 결과를 통해 본 발명자들은 미선나무를 이용한 골질환의 치료제로서 미선나무의 잎의 사용이 가장 유용함을 알 수 있었다. 이에 본 발명자들은 미선나무 잎을 대상으로 추출물을 수득하는 과정에서 용매를 달리하여 수득한 추출물에 대하여 파골세포 분화 억제 활성을 분석하였다.From the results of Example 2, the present inventors found that the use of leaves of Miseon tree as a treatment for bone disease using Miseon tree is the most useful. Accordingly, the present inventors analyzed the osteoclast differentiation inhibitory activity of the extract obtained by changing the solvent in the process of obtaining the extract from Miseon tree leaves.
구체적으로 상기 실시예 1의 과정에서, 미선나무 잎에 대하여 메탄올 이외에 다른 용매인 에틸아세테이트, 헥산 및 부탄올을 각각 사용하여 각 용매별 미선나무 잎 추출물을 수득하였고, 수득한 각 용매별 추출물에 대하여 상기 실시예 2의 실험과 동일하게 파골세포 분화 억제 실험을 수행하였다. Specifically, in the process of Example 1, ethyl acetate, hexane, and butanol, other solvents other than methanol, were respectively used for Miseon tree leaves to obtain Miseon tree leaf extracts for each solvent. In the same manner as in Example 2, an osteoclast differentiation inhibition experiment was performed.
그 결과, 도 3에 나타낸 바와 같이, 에탄올, 에틸아세테이트, 헥산 및 부탄올을 이용하여 수득한 미선나무 잎 추출물에 비해 미선나무 잎의 메탄올 추출물이 가장 우수한 파골세포 분화 억제 활성이 있는 것으로 나타났다.As a result, as shown in FIG. 3 , it was found that the methanol extract of Miseon tree leaf had the best osteoclast differentiation inhibitory activity compared to the Miseon tree leaf extract obtained using ethanol, ethyl acetate, hexane and butanol.
따라서 효과적인 파골세포의 분화억제를 위해서는 추출 용매로써 메탄올을 선택하여 사용하는 것이 매우 중요하다는 것을 알 수 있었다. Therefore, it was found that it is very important to select and use methanol as an extraction solvent for effective inhibition of differentiation of osteoclasts.
이제까지 본 발명에 대하여 그 바람직한 실시예들을 중심으로 살펴보았다. 본 발명이 속하는 기술 분야에서 통상의 지식을 가진 자는 본 발명이 본 발명의 본질적인 특성에서 벗어나지 않는 범위에서 변형된 형태로 구현될 수 있음을 이해할 수 있을 것이다. 그러므로 개시된 실시예들은 한정적인 관점이 아니라 설명적인 관점에서 고려되어야 한다. 본 발명의 범위는 전술한 설명이 아니라 특허청구범위에 나타나 있으며, 그와 동등한 범위 내에 있는 모든 차이점은 본 발명에 포함된 것으로 해석되어야 할 것이다.So far, with respect to the present invention, the preferred embodiments have been looked at. Those of ordinary skill in the art to which the present invention pertains will understand that the present invention may be implemented in a modified form without departing from the essential characteristics of the present invention. Therefore, the disclosed embodiments are to be considered in an illustrative rather than a restrictive sense. The scope of the present invention is indicated in the claims rather than the foregoing description, and all differences within the scope equivalent thereto should be construed as being included in the present invention.
Claims (10)
상기 추출물은 파골세포의 분화 또는 형성을 억제하는 것을 특징으로 하는, 골다공증, 파세트 골질환, 구루병, 골연화증, 신부전 환자의 신성골이영양증, 류머티스성 골질환 및 퇴행성 골질환으로 이루어진 군 중에서 선택되는 골질환의 예방 또는 치료용 약학적 조성물.The method of claim 1,
The extract is characterized in that it inhibits the differentiation or formation of osteoclasts, osteoporosis, facet bone disease, rickets, osteomalacia, renal osteodystrophy of renal failure patients, rheumatoid bone disease and bone selected from the group consisting of degenerative bone disease A pharmaceutical composition for preventing or treating a disease.
상기 추출물은 파골세포의 분화 또는 형성을 억제하는 것을 특징으로 하는, 골다공증, 파세트 골질환, 구루병, 골연화증, 신부전 환자의 신성골이영양증, 류머티스성 골질환 및 퇴행성 골질환으로 이루어진 군 중에서 선택되는 골질환의 예방 또는 개선용 건강기능식품.7. The method of claim 6,
The extract is characterized in that it inhibits the differentiation or formation of osteoclasts, osteoporosis, facet bone disease, rickets, osteomalacia, renal osteodystrophy of renal failure patients, rheumatoid bone disease and bone selected from the group consisting of degenerative bone disease Health functional food for preventing or improving disease.
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| KR102043354B1 (en) * | 2017-10-17 | 2019-11-12 | 주식회사 프로셀컴퍼니 | Composition for Prophylaxis and Treatment of Osteoporosis Comprising Abeliophyllum Distichum Extract |
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