KR101954695B1 - A pharmaceutical composition for treating or prevebting bon disease - Google Patents

Abstract

The present invention relates to a pharmaceutical composition for preventing or treating bone diseases in which therapeutic adverse effects are significantly reduced by using any one or more natural extracts selected from the group consisting of rosewood plant extracts, lamellar plant extracts, mulberry plant extracts and horseradish extracts, And a food composition for improving or alleviating bone diseases, wherein the composition effectively inhibits differentiation into osteoclasts, thereby effectively preventing or treating bone diseases.

Description

FIELD OF THE INVENTION The present invention relates to a pharmaceutical composition for preventing or treating bone diseases,

The present invention relates to a plant or a plant selected from the group consisting of (a) at least one plant selected from the Rosales plant group consisting of a rock horse chestnut, a plum horsetail, a mustache, a cherry tree, a radish tree, a mountain cherry tree, an apricot tree, Plant extracts; (b) one or more plant extracts selected from the group consisting of bear plants, dogwood trees, dogwood trees, white-tailed trees, cornus and corn plant; (c) one or more botanical extracts selected from the group of Boehmeria plants consisting of anchovy and turtle tail; And (d) one or more plant extracts selected from the group consisting of Carpinus plants consisting of aphids and aphids, as an active ingredient. ≪ / RTI >

The osteoclast, which is a polynuclear large cell, has a function of destroying and absorbing bone tissue, and is said to be responsible for destroying the bone matrix and degrading the bone mineral. Activated osteoclasts have three or more nuclei. To differentiate into osteoclasts from mature osteoclast precursor cells, M-CSF (macrophage colony stimulating factor) and RANKL (receptor activator of nuclear factor-kappa B ligand) (Mojtaba A., et al., Cancer Biol Ther., 7: 1,3-9; 1 (2008)). The osteoclasts cause abnormal bone tissue destruction and absorption due to imbalance with osteoblasts in the bone, resulting in osteoporosis in which bone mass and bone density are decreased, osteoporosis in bone, It is the cause of osteomalacia in which lime is removed, fibrous osteitis in which bone marrow is fibrous, periodontitis in which alveolar bone is lost, rheumatoid arthritis which causes joint destruction and deformation It is known.

Although the mechanism of the diseases is not clearly understood, the imbalance between osteoclasts and osteoblasts may be caused by various causes such as bone metastasis of cancer cells, endocrine disorders, hyperparathyroidism, A variety of drugs and treatments capable of effectively inhibiting osteoclast destruction and absorption by osteoclasts have been actively studied. Currently, bisphosphonate-based therapeutic agents such as Fosamax (aledronate), Actonel (ingredient name: risedronate), Zometa (ingredient name: zoledronate) and the like are widely used for treatment of bone damage by osteoclasts such as osteoporosis However, most of these agents induce the death of osteoclasts to delay or inhibit the loss of bones. Therefore, various kinds of side effects such as osteonecrosis, severe atrial fibrillation, inability to bones or joints, and pain in the musculoskeletal system (Coleman RE., Br J Cancer, 98: 1736-1740 (2008)).

On the other hand, cancer cells that have metastasized to the bone from breast cancer and prostate cancer also promote osteoclast formation, resulting in serious bone diseases.

Therefore, there is a need to develop medicines which can overcome the disadvantages of the existing bisphosphonates.

One object of the present invention is to improve the disadvantages of existing bisphosphonates by using one or more natural extracts selected from the group consisting of rosemary plant extracts, lamellar plant extracts, mulberry plant extracts and horseradish extracts, And to provide a pharmaceutical composition for preventing or treating osteopathy which is significantly reduced.

However, the technical problem to be solved by the present invention is not limited to the above-mentioned problems, and other matters not mentioned can be clearly understood by those skilled in the art from the following description.

The present inventors have found that when the extract of any one or more selected from the group consisting of rosemary plant extract, lamellar plant extract, mulberry plant extract and horseradish extract is used as an effective ingredient, The present invention has been completed.

The term " osteoclast " refers to a cell that performs a function of destroying a bone matrix and degrading minerals of a bone, thereby performing tissue destruction and absorption. It has been reported that the osteoclast can cause osteopathy when imbalance with osteoblasts occurs in the bone. Therefore, for the purpose of the present invention, the bone disease may be selected from the group consisting of osteoporosis, osteomalacia, rickets, osteopenia, fibrotic bone disease, osteoarthritis, osteoarthritis, bone atrophy, paget's disease, periodontitis, rheumatoid arthritis rheumatoid arthritis, metabolic bone disease, and bone damage caused by bone metastasis of cancer cells. However, it is not limited to any disease that can be induced by osteoclast tissue destruction and bone resorption No.

In one embodiment of the present invention, the present invention provides a method for producing roses, comprising the steps of: (a) preparing a roses comprising rosemary spikelets, plum spikelets, anchovy, cherry, radish, cherry, apricot, cherry, cherry, At least one plant extract selected from the group of plants; (b) one or more plant extracts selected from the group consisting of bear plants, dogwood trees, dogwood trees, white-tailed trees, cornus and corn plant; (c) one or more botanical extracts selected from the group of Boehmeria plants consisting of anchovy and turtle tail; And (d) one or more plant extracts selected from the group consisting of Carpinus plants consisting of aphids and aphids, as an active ingredient. Lt; / RTI >

Here, the rock sparrow (tree) is a deciduous shrub that is one of the seed breeding plants, and is known to grow in rock crevices at the foot of a mountain. It is about 1m high, has small hairs on the hairs, and the old branch is grayish brown. The opposite hairs are 3 ~ 6cm long ovate and have serrate on the edge. It blooms in April to May, and flowers are white. It can be planted for ornamental purposes and can be used as a raw material. Preferably, the extract may be obtained by using at least one of leaves or flowers of the rock spikelets. More preferably, the name of the rock spikelets is Deutzia grandiflora var. Baroniana Diels. < / RTI >

Also, the plum blossom (tree) is also called dangganggok, and it grows in a rock gap in the mountain. About 1m high, the bark is ash-colored and irregularly peeled off. Leaves are opposite, oval or wide, with serrate on the edge, 4 ~ 6cm in length on both sides. Flowers bloom in May, white and 1 ~ 3 on old branches. One or two leaves under the flowers may also be running. The calyx tube is attached to the ovary and the Virgin Mary is closely packed with the small peduncle. Preferably, the extract may be obtained by using any one or more of the above-mentioned plum blossom foliage or flowers, and more preferably, the scientific name of the plum blossom foliage may be Deutzia uniflora Shirai, but is not limited thereto.

In addition, the above-mentioned petiole (tree) grows on both sides of the foot of the mountain and around the village, and the height is 5 to 10 meters. The bark is cork developed, the small branch is brown, the leaves are slanting, wide ovate, pointed end, and irregular double sawtooth. The length is 5 ~ 12cm, the back side of the leaf is green, and the petiole is 2 ~ 3cm long. Flower is pale red or white and blooms one by one in April-May before leaf, small peduncle is 8mm long, calyx is oval, and has serrate-like sawtooth. Petal is round and length is 10 ~ 12mm. The pistil is about 15mm long, similar to surgery, and the stigma has a cup shape. Ovaries are hairy. The fruit is egg-shaped as a nucleus and it has a sweet taste and ripens yellow in July-August. Preferably, the extract can be obtained by using at least one of leaves, stalks, bark, fruits, and roots of the auricula, more preferably, Prunus mandshurica var. glabra. < / RTI >

In addition, the cherry tree is a deciduous broad-leaved arboreous tree, and its origin is Korea, and it is distributed in Japan and China and inhabits in mountainous regions. Its height is 10 ~ 20m and it is peeled sideways with dark brown. Leaves are alternate phyllotaxis, egg-shaped or egg-shaped, 6 ~ 12cm long, with small mounts on the edge. The back side of the leaf is grayish green. Flowers bloom in April or May in pink or white with arthropods or phloem phloem, with flower stalks, with no calyx hairs. Fruit is a nucleus and ripens in June to July, and is round. Preferably, the extract can be obtained by using at least one of leaves, stems, barks, fruits and roots of the cherry tree, more preferably the name of the cherry tree is Prunus serrulata var. spontanea (Maxim.) E.H. Wilson, but is not limited thereto.

In addition, here, the radiant tree (peach tree) belongs to Rosaceae, which is arboreal deciduous and watery, about 6 m in height, has no hairs on small branches, and has hair on winter buds. Leaves are alternate, lanceolate, obovate, and lanceolate, 8 ~ 15㎝ long, 1.5 ~ 3.5㎝ wide, with no hairs on both sides and with dull serrated edges. Flower is pale red, blooms earlier than leaves in April to May, and fruit is ovary, which is an ovate-like form, ripened in August-September. Preferably, the extract may be obtained using at least one of leaves, stalks, bark, fruits, and roots of the radiant tree, more preferably, the scientific name of the radiant tree may be Prunus persica (L.) BATSCH , But is not limited thereto.

In addition, the above-mentioned mountain cherry tree is a deciduous broad-leaved bark tree growing in mountain forests of the whole country, and its stem reaches 20m in height. The stem skin is dark brown, peeled sideways, and the lobes are long side. Leaves are alternate, oval or obovate. The petiole has no hairs and purplish red, and there is a pair of honey glands on the upper part. Flowers bloom in April to May in white or pale red, 2-3 or so to form phloem. The petals are round and have no fragrance. Peduncle, operating table, style, ovary hairless. The fruit is nuclear and ripens in black-purple in June-June. It is distributed worldwide in Japan, Russia Sakhalin region. Preferably, the extract may be obtained by using at least one of leaves, stems, bark, fruits, and roots of the above-mentioned cherry tree, more preferably the scientific name of the cherry tree is Prunus sargentii Rehder, It is not.

Here, the apricot tree is a deciduous tall tree planted near the house, about 5 m in height, and the bark is reddish brown, and the hawk branch is reddish brown. Leaves are alternate wide oval or broad ovate. Flowers bloom in April with a light red color. Calyx leaves are 5, reddish purple and tilted backward. There are 5 petals and a round shape. The fruit is a round nucleus, about 3cm in diameter, and ripens in yellow or reddish yellow in July. They plant in the southern central region of Korea. It is planted as ornament for Chinese origin, the fruit is edible or medicinal, and the wood is used as an appliance material. Preferably, the extract can be obtained by using at least one of leaves, stalks, bark, fruit, and roots of the apricot tree, more preferably the apricot tree is Prunus armeniaca var. ansu Maxim, but is not limited thereto.

In addition, here, the above-mentioned cherry tree grows in the forest, its height is about 10m, and the bark is gray-brown. Leaves are alternate phyllotaxis, long oval, narrowed at both ends, with small sawtooth on the edge and hairs on the vein axillary. Petiole is 8 ~ 12cm. The flower blooms in May and is white, and runs on a gun inflorescence grown from an old branch. Calyx is short, dull end, teeth-like sawtooth. The petals are 5, oval, shorter than the surgery, and have no hairs in the ovary. Fruits are round with nucleus and ripen in black purple in July, and calyx and stamen remain intact. It is widely distributed in Korea (Jeju) and Japan. Preferably, the extract may be obtained using at least one of the leaves, stems, barks, fruits, and roots of the senberry cherry tree. More preferably, the scientific name of the senberry cherry tree may be Prunus buergeriana Miq. It is not.

In addition, the island cherry tree is one of the Ulleungdo-specific plants and its height reaches 20m. The bark is gray-brown and the small branches are grayish brown or gray-purple and have no hairs. The winter snow is long oval and has no hairs. Leaves are alternate phyllotaxis, ovate or ovate, long pointed end, lower heart or base, 8 ~ 15㎝ long, 4 ~ 9㎝ wide, no hairs on both sides. The surface is green, the back side is gray green, and the sawtooth is pointed like hair. The petiole is 2.5 ~ 3㎝ long with no hairs, yellowish green and has a nectar at the end. Flowers bloom later than leaves in April and have a diameter of 2.5㎝. There are no peduncle and 2 ~ 5 flowers are arranged in umbrella shape. Small peduncle is 15 ~ 18㎜ long and has no hair. The calyx leaf is backward tilted, has no sawtooth, and has a length of about 5mm and has a triangular shape. The calyx tube is 6 ~ 7㎜ in length. The petals are obovate or wide elliptical, 12 ~ 13㎜ long, and the ends are pierced. Surgery is 30 ~ 40, length is 5 ~ 10㎜, pistil is hairless and length is about 1㎝. The fruit is round or ovate, pointed or rounded, 1 ~ 2㎝ in length, 7㎜ in width, and rips in red in June. The bovine suture is durable, 2㎝ in length, and 2mm in diameter. Preferably, the extract can be obtained using at least one of the leaves, stems, barks, fruits and roots of the island cherry tree, more preferably the scientific name of the island cherry tree is Prunus takesimensis NAKAI. It is not.

In addition, here, the above-mentioned cherry tree reaches a height of 10 m, the bark is grayish brown, and the small branches have hairs. Leaves are alternate phyllotaxis or oval, with sawtooth on edge, hairs with petiole at first. There is hair on the back side of leaf, and sawtooth. Flowers bloom earlier than leaves in April, light pink and 2 ~ 5 flowers bloom in mountain-shaped inflorescence. Small flower stalks are hairy. The bottom of the calyx tub is round and swollen. The petal is elliptical, concave at the end, 10 ~ 12mm in length, and the hair is denser at the bottom of the pistil. The fruit is round and ripens black from June to July. It is distributed in Korea (Jangsan-goat, Wibongsan, Jiri-san, Bogil-do, Jeju) and Japan. Preferably, the extract may be obtained by using at least one of the leaves, stems, bark, fruits and roots of the above-mentioned cherry tree, more preferably, Prunus pendula for. but are not limited to, ascendens.

In addition, the Yoshino cherry tree is a plant belonging to the genus Rosaceae (Prunus L.). It grows in the temperate zone of the northern hemisphere and is known to be able to eat its fruit or bark. Preferably, the extract is obtained by using the leaves of the Prunus yedoensis Matsum, but is not limited thereto

In addition, the above-mentioned alder tree is a deciduous arboreous tree that grows in the foothills of the southern central region and leaves. The growth environment is sunny, grows in a lot of low-tillage soil, height is 3 ~ 8m, the leaves are egg or round, and the edges are wavy. The flower has no peduncle, 20 ~ 30 branches at the end of small branch, heading toward the sky, 3 ~ 8㎝ in length, 2 ~ 3㎝ in butterfly, white or white green, similar to petal shape. The nuts are rounded in red in October, and the shell surrounding the seeds is meaty and edible. It may contain isoquercitrin and gallic acid. It is used for ornamental and industrial purposes. The fruit is used for edible and medicinal purposes. Preferably, the extract can be obtained by using the leaves of the above-mentioned cornstalks, more preferably the cornstarch can be Cornus kousa F.Buerger ex Miquel, but is not limited thereto.

In addition, here, the bear's name is also referred to as a ' It grows at the foot of a mountain or at the edge of a forest and is more than 15 meters high. The bark is grayish brown and cracks vertically irregularly. Leaves are 8 ~ 18cm long, green, facing, oval or wide ovate. There are small hairs on the front side of the leaves and a lot of white hairs on the back side. The petiole is 1 ~ 3cm long and has no hairs. From June to August, the four-branch flower from the tip of the twig is gathered closely to the flower of the saccharide. Inflorescence is 8 ~ 14cm in diameter and has 1 pistil and 4 stamens. The peduncle is 3 ~ 4cm long and has no hairs. The petals are wide and narrow. The fruit is round and about 6mm in diameter, ripened in October with a bluish black color. Seeds are round and have many concave spots. It is also used as a gardening tool, and wood is used as a craft material. It is distributed in Korea, Japan, Taiwan and China. Preferably, the extract can be obtained by using any one or more of the leaf, stem, bark, fruit, and root of the bear, more preferably the scientific name of the bear is Cornus macrophylla. However, It is not.

Also, the dogwood grows in the valley, and the height is about 10 m. The bark is blackish brown and cracks like a net. Leaves are opposite, broad ovate or elliptical, with slightly wavy hair on both sides, plain edges and 4-5 pairs of side veins. Length 5 ~ 8cm, butterfly 3 ~ 5cm, the back is white. Petiole is 1 ~ 3cm long. Flowers bloom in May-June, white, hanging in the cyme inflorescence. The peduncle is 1.5 ~ 2.5cm in length and petals are small. The pistil is shorter than the surgery and the operating table is similar to the petal length. The fruit is round as a nucleus and has a diameter of 6-7mm and ripens black in September-October. It is planted as a garden, and wood is used as a building material or an appliance material. It is distributed in Korea and China. Preferably, the extract may be obtained using at least one of leaves, stalks, bark, fruits, and roots of the tree, more preferably, Cornus walteri may be a scientific name of the tree. However, no.

Here, the stratum corneum is grown in the valley forest of the mountain. It reaches a height of 20m, and the branches spread horizontally running to the stratum. Small branches are colored with deep reddish purple in winter, and water is flowing when branches are cut in spring. Leaves are alternate, wide oval, pointed end. Leaf edges are flat, side veins are 5 to 8 in number, petiole is red, and leaf back is white. There are fine hairs on both sides of leaf. Flowers bloom from May to June, white, and form a flower bed. The petals are wide and bloom with calyxes and hairs on the outside. There are 4 stamens, anthers are T-shaped, and one pistil. The fruit is round and round in black color. It is distributed in Korea, Japan and China. Preferably, the extract may be obtained using at least one of leaves, stalks, bark, fruits, diseases and roots of the stratum, and more preferably, the stratum may be a Cornus controversa, It is not.

In addition, the above-mentioned white wood is also called red mulberry tree. It grows in mountainous waters, and its height is about 3m. The bark is red, the bone is white and the young branches have no hair. Leaves are opposite, oval or oval, 5 ~ 10cm long and 3 ~ 4cm wide. The tip is pointed, the bottom is round or wide wedge shape, and the edge is flat. The surface is green, with loose hairs and white on the back. The petiole is 6 pairs and petiole is 1 ~ 2.5cm long and has no hairs. Flowers bloom in May to June with yellowish-white color, hanging in the form of an umbrella flower. The inflorescence is 4 ~ 5cm in diameter and the peduncle is 5 ~ 10mm in length. The calyx is divided into 4 branches, and the branches are pointed and short. Petals are 4, egg-shaped, with a length of about 3mm. Four stamens are similar in length to petals, pistils are shorter than stomachs, and ovaries are in the lower part. Fruit is elliptical nucleus and white or blueish white, ripened from August to September. The seeds are narrow and flat at both ends. They breed with seeds and bones. It is planted in a garden with excellent ornamental value. It also has anti-inflammatory and hemostatic effects on the bark and leaves, so it is also used as medicinal herb. It is distributed in Korea (South Pyongan, Pyeongan, North Hamgyong, North Hamgyong), China, Sakhalin, Mongolia, Siberia and Europe. Preferably, the extract may be obtained by using at least one of leaves, stalks, bark, fruit, diseases and roots of the above-mentioned P. lanceolata. More preferably, the name of the stratum may be Cornus alba, It is not.

In addition, the above-mentioned corn oil is the fruit of the cornstalks which is a deciduous tree with the stratum corneum. Elliptical nuclei are green at first, but ripen reddish from October to October. Seeds are long oval and have ridges. It has a bitter sweetness and strong acidity. It is harvested after the frost fall in mid-October. The flesh is separated from the seeds and the meat is used as a material for sake, tea and Chinese medicine. The pulp contains the glycosides such as cornin, morroniside, loganin, tannin and saponin and organic acids such as wine, malic and tartaric acid. In addition, it contains vitamin A and a large amount of sugars. Seeds include palmitic acid, oleic acid, linoleic acid, and the like. Among the components, cornin is known to have an excitatory action of parasympathetic nerves. From the past, the pulp was weakened in one batch. Sansui is native to Korea and China, and is planted in the middle of South Korea. In Korea, Shandong, Gyeonggi Province, Gyeonggi Province, Gyeonggi Province, and Gyeongsangbukdo, Yusung Gun, and other local products are shipped every year. Preferably, the extract may be obtained by using at least one of leaves, stalks, bark, fruit, diseases and roots of the above-mentioned corn oil, more preferably the corn tree may be Cornus officinalis, no.

In addition, the above-mentioned dogs are perennial plants, growing mainly in the vicinity of the sea, and about 1 meter in height. The stem is gathered and the leaf is opposite, the leaf is ovoid, with regular sawtooth on the edge. Flowers bloom in July-October with a matured abandon, light green, and bloom on the axilla. Fruit is achene, obovate, white hairs on the upper part. Preferably, the extract can be obtained by using at least one of outpox, leaf, stem, bark, fruit, disease, and root of the canola, more preferably, Boehmeria platanifolia Franch. et Sav, but is not limited thereto.

Here, the tortoise tail is 50-100 cm high, and the stem is reddish brown but sometimes it is greenish brown. It has hair but gradually disappears and branches are thin. Leaves are opposite to each other and are egg-shaped, 4 ~ 8cm long, 2.5 ~ 4cm wide, with long tail like tail and 5 ~ 6 large sawtooth on edge. There are hairs on the front, hairs on the veins on the back, and petiole length is 1 ~ 3cm, and it is reddish. The flower is one male and one female, but sometimes there is a dandelion, and it blooms from July to August. The female flower is running on the axilla of the lower part of the stem. The female flower stamens are on the axilla of the upper part of the stem. Male flower has four foliage pieces and surgery. Several female flowers run together. There is one ovary in the tubular flower shape, and one style. Fruit is aquatic, ripened in October, and is a long egg shape, but several are gathered together and appear as several, with hair at the end of the fruit. Korea is native to Korea, China, and Japan. It lives near the valley of the valley and at the edge of the forest. Preferably, the extract can be obtained using at least one of outposts, leaves, stems, bark, fruits, diseases and roots of the turtle tail, more preferably, the oecheria spicata (Thunb.) Thunb But is not limited thereto.

In addition, here, the above-mentioned trees grow in the forest below the hillside. Height is about 15m and diameter is about 70cm. The bark is ash-colored and rugged but does not burst, and white lobes are scattered. Leaves are elliptical or long egg-shaped, with 2 rows of phyllotaxis, with pointed ends, thin sawtooth, 4 ~ 8cm long. The vein is 12 ~ 15 pairs in length and has hair on the front and back veins. The petiole is 4 ~ 12mm long and hairy. Flowers bloom in May to May as a bloom. The male flower ear does not have a base, it runs on the back, and the female flower ear has a base and is attached sparsely. The fruit is nuts, which are contained in the perianths and have a broad egg shape and ripen in October. The wood is dense, hard and elastic. It is used for equipment, wood, woodwork, letterpress, wood for mushroom cultivation, etc. It is distributed in Korea (Jeonnam, Jeonbuk, Gyeongnam), Japan, and China. Preferably, the extract can be obtained by using at least one of the leaves, stalks, bark, fruits, diseases and roots of the above-mentioned cervical spine, more preferably the carpinus tschonoskii is the scientific name of the cervical spine But is not limited to.

In addition, here, the cedar tree is also called a standing tree, and the origin of the tree name is not certain, but it is assumed to be a tree in the west. It grows mainly in mountainous areas and is commonly found. Seedwood is used as an alley for cultivation of shiitake mushroom, but its production is less than that of oak and it is widely used for firewood. The types of trees that are similar in appearance are dogwood, magpie, and sosa. The height is 15m and the diameter is 1m. The bark is gray and bumpy like muscles. Leaves are alternate phyllotaxis, 5.5 ~ 7.5cm long elliptical or long ovate, pointed end with double sawtooth on edge, hairs on back vein. The flower is a male and female, blooming in May and running in unspecified inflorescence. The male flower sprout hangs on the small branches and sags downward. The male flower rides one by one in each bubble, has 8 operations, and the operating table is divided into 2 pieces. There is a canopy of female flower ear, two female flowers in each flower, and two pistils. Females have long cylindrical shape, 4 ~ 8cm in length, with sagging downward, 4 ~ 8 pods, with deeply pitted teeth on one side and one protrusion on the opposite side. Fruit is egg yolk with 3mm long egg shape and ripen in October. It is distributed in Korea (south of the Yellow Sea), Japan, and China. Preferably, the extract may be obtained using at least one of the leaves, stems, bark, fruits, diseases and roots of the cirrus trees. More preferably, the name of the cirrus can be Carpinus laxiflora, It is not.

In one embodiment of the present invention, the pharmaceutical composition according to the present invention may inhibit the differentiation into osteoclast.

The extract according to the present invention preferably inhibits the activity of a macrophage colony stimulating factor (M-CSF) and / or a receptor activator of nuclear factor-kappa B ligand (RANKL) required for differentiation of osteoclast precursor cells To inhibit osteoclast differentiation. However, there is no limit as long as it can be used for prevention and treatment of osteopathy by inhibiting osteoclast differentiation.

The above-mentioned " macrophage colony stimulating factor (M-CSF) " is a cytokine that expresses osteoblasts and bone marrow stromal cells, plays an important role in osteoclast formation and mainly plays a role in cell proliferation, survival, cytoskeletal organization (Kim SY, et al., J Korean Orthop Assoc., 44: 151-158 (2009)).

In addition, the "RANKL (receptor activator of nuclear factor-kappa B ligand)" is a factor expressing osteoblast cells, which adhere to RANKL receptors in osteoclast precursor cells to induce osteoclast growth and differentiation (Mojtaba A., Cancer biology & Therapy, 7: 1,3-9; 1 (2008)). RANKL promotes osteoclast differentiation by activating transcription factors such as c-fos, NFATc1 (nuclear Factor Of Activated T-Cells, cytoplasmic, calcineurin-dependent 1) and NF-kB (nuclear factor kappa B) (LEE ZH., et al., Biochem Biophys. (2002)). In addition, it is known that activation of signal transduction such as phosphatidylinositol 3-kinase and extracellular signal-regulated kinase (ERK) plays a role in promoting osteoclast survival and function Res Commun., 305: 211-213 (2003)).

In another embodiment of the present invention, a) a Rosales plant group consisting of a rock horse chestnut, a plum horsetail, an apricot, a cherry tree, a radiant tree, a mountain cherry tree, an apricot tree, a cherry tree, a cherry tree, ; (b) Cornus flora, consisting of the bear's words, dogwood, dogwood, white-tailed wood, cornus and corn oil; (c) Plants of the Boehmeria family, consisting of the dogs and turtle tail; And (d) a Carpinus plant group consisting of an open-top tree and a perennial tree, and pulverizing the same; B. Extraction of the powder with an organic solvent in a solvent extraction device or an Accelerated Solution Extractor followed by filtration, and a method for producing a pharmaceutical composition for preventing or treating bone diseases.

In one embodiment of the present invention, the temperature of the solvent extraction apparatus may be 30 ° C to 60 ° C, preferably 50 ° C, but is not limited thereto. When the temperature is below 30 ° C, the temperature is not sufficiently raised and the active ingredient can not sufficiently dissolve in the organic solvent. If the temperature is higher than 60 ° C, the components of the extract may be denatured.

In another embodiment of the present invention, the organic solvent used in the solvent extraction apparatus may be ethanol or methanol, preferably ethanol, but is not limited thereto. In the case of using ethanol in the present invention, there is an advantage that the yield of the active ingredient contained in the extract can be further improved as compared with the case of using methanol.

In another embodiment of the present invention, the method may further comprise the step of concentrating the filtered filtrate. The step of concentrating the filtrate may be a step of concentrating using a vacuum concentrator or a vacuum concentrator, but the present invention is not limited thereto, so long as the extracted components can be concentrated.

The pharmaceutical composition of the present invention can further be administered in combination with other anticancer drugs, thereby effectively inhibiting general cancer cell proliferation and cancer metastasis, thereby treating bone damage caused by cancer and treating cancer at the same time.

Examples of the anticancer agent include nitrogene mustard, imatinib, oxaliplatin, rituximab, elotinib, neratinib, lapatinib, zetitib, bandetanib, nilotinib, semathanib, conservinib, acitinib, , Corticosteroids, cisplatin, cetuximab, bismuth alum, asparaginase, tretinoin, hydroxycarbamate, terephthalic acid, tretinoin, Amlodipine, amlodipine, amlodipine, amlodipine, amlodipine, amide, amlodipine, amlodipine, amlodipine, amlodipine, But are not limited to, chitosan, gemcitabine, doxifluridine, femetrexed, tegafur, capecitabine, gimeracin, oteracil, azacytidine, methotrexate, uracil, cytarabine, fluorouracil, Tabin, flutamide, But are not limited to, paclitavine, decitabine, mercaptopurine, thioguanine, cladribine, calmophor, ralitriptycde, docetaxel, paclitaxel, irinotecan, velotecan, topotecan, vinorelbine, , Mitomycin, bromomycin, dactinomycin, pyra rubicin, aclarubicin, pepromycin, temsirolimus, temozolomide, epidermal, iopospermide, cyclophosphamide, melphalan, altret But are not limited to, corticosteroids, corticosteroids, corticosteroids, corticosteroids, corticosteroids, corticosteroids, corticosteroids, corticosteroids, At least one selected from the group consisting of tamoxifen, toremifene, testolactone, anastrozole, letrozole, borozol, bicalutamide, rosmutin, 5FU, borinostat, entinostate and carmustine may be used However, The.

In the present invention, the pharmaceutical composition may be in the form of capsules, tablets, granules, injections, ointments, powders or beverages, and the pharmaceutical composition may be a human.

The pharmaceutical composition of the present invention may be formulated in the form of an oral preparation, an external preparation, a suppository, and a sterile injection solution in the form of powders, granules, capsules, tablets, aqueous suspensions, etc., have. The pharmaceutical composition of the present invention may comprise a pharmaceutically acceptable carrier. The pharmaceutically acceptable carrier may be a binder, a lubricant, a disintegrant, an excipient, a solubilizing agent, a dispersing agent, a stabilizer, a suspending agent, a coloring matter, a perfume or the like in the case of oral administration. A solubilizing agent, an isotonic agent, a stabilizer and the like may be mixed and used. In the case of topical administration, a base, an excipient, a lubricant, a preservative and the like may be used. Formulations of the pharmaceutical compositions of the present invention may be prepared in a variety of ways by mixing with pharmaceutically acceptable carriers as described above. For example, oral administration may be in the form of tablets, troches, capsules, elixirs, suspensions, syrups, wafers, etc. In the case of injections, they may be formulated in unit dosage ampoules or in multiple dosage forms have. Other, solutions, suspensions, tablets, capsules, sustained release formulations and the like.

Examples of suitable carriers, excipients and diluents for formulation include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltoditol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, Cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate or mineral oil. Further, it may further include a filler, an anticoagulant, a lubricant, a wetting agent, a flavoring agent, an emulsifying agent, an antiseptic, and the like.

The route of administration of the pharmaceutical compositions according to the present invention may be, but is not limited to, oral, intravenous, intramuscular, intraarterial, intramedullary, intrathecal, intracardiac, transdermal, subcutaneous, intraperitoneal, intranasal, Sublingual or rectal. Oral or parenteral administration is preferred. The term "parenteral" as used herein includes subcutaneous, intradermal, intravenous, intramuscular, intraarticular, intrasynovial, intrasternal, intrathecal, intralesional and intracranial injection or infusion techniques. The pharmaceutical compositions of the present invention may also be administered in the form of suppositories for rectal administration.

The pharmaceutical composition of the present invention varies depending on various factors including the activity of the specific compound used, age, weight, general health, sex, diet, time of administration, route of administration, rate of excretion, drug combination and the severity of the particular disease to be prevented or treated. And the dose of the pharmaceutical composition may be appropriately selected by a person skilled in the art depending on the condition of the patient, the body weight, the degree of disease, the type of drug, the route of administration and the period of time, and may be 0.0001 to 50 mg / kg or 0.001 to 50 mg / kg. The administration may be carried out once a day or divided into several times. The dose is not intended to limit the scope of the invention in any way. The pharmaceutical composition according to the present invention can be formulated into pills, dragees, capsules, solutions, gels, syrups, slurries and suspensions.

Another embodiment of the present invention provides a method for treating bone diseases using the pharmaceutical composition.

In another embodiment of the present invention, there is provided a plant of Rosales plant consisting of (a) Rosaceae, Plum Horseradish, Anchovy, Cherry, Radish, Mountain Prunus, Apricot Tree, Prunus Prunus, At least one plant extract selected from the group consisting of: (b) one or more plant extracts selected from the group consisting of bear plants, dogwood trees, dogwood trees, white-tailed trees, cornus and corn plant; (c) one or more botanical extracts selected from the group of Boehmeria plants consisting of anchovy and turtle tail; And (d) one or more plant extracts selected from the group consisting of Carpinus plants consisting of aphids and hemlines, as an active ingredient, for improving or alleviating bone diseases, ≪ / RTI >

As used herein, the term " food composition " is used variously for improving and preventing bone diseases. The food composition containing the composition of the present invention as an active ingredient may be used in various foods such as beverages, gums, tea, vitamins Granules, tablets, capsules, confections, rice cakes, breads, and the like. Since the food composition of the present invention is modified and prepared from an existing food-grade food having little toxicity and side effects, it can be safely used for prolonged use for preventive purpose. When the composition of the present invention is contained in the food composition, the amount thereof may be added in a proportion of 0.1 to 100% of the total weight. Here, when the food composition is prepared in a beverage form, there are no particular limitations other than those containing the food composition at the indicated ratios and may contain various flavors or natural carbohydrates such as ordinary beverages as an additional ingredient. That is, natural carbohydrates include monosaccharides such as glucose, disaccharides such as fructose, sucrose and the like, and conventional sugars such as polysaccharide, dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol and erythritol can do. Examples of the flavors include natural flavors (such as tau martin, stevia extract (for example, rebaudioside A and glycyrrhizin), and synthetic flavors (for example, saccharin and aspartame). The food composition of the present invention can be used as a food composition containing various nutrients, vitamins, minerals (electrolytes), flavors such as synthetic flavors and natural flavors, colorants, pectic acid and its salts, alginic acid and its salts, Stabilizers, antiseptics, glycerin, alcohols, carbonating agents used in carbonated beverages, etc. These components may be used independently or in combination. The ratio of such additives is usually in the range of 100 parts by weight per 100 parts by weight of the composition of the present invention But is not limited to, 0.1 to 100 parts by weight.

The present invention relates to a method for inhibiting osteoclast differentiation using a pharmaceutical composition comprising at least one natural extract selected from the group consisting of rosemary plant extracts, lamellar plant extracts, mulberry plant extracts and horseradish extracts as active ingredients It is possible to prevent or treat bone diseases effectively with low side effects.

FIG. 1 is a view showing the result of confirming toxicity of a rock horse chestnut and its plant extracts on cells according to an embodiment of the present invention. FIG.
FIG. 2 is a view showing the result of confirming toxicity of a cornus and its herbal extracts on cells according to an embodiment of the present invention. FIG.
FIG. 3 is a graph showing the results of examining toxicity of the extract of the canola and its plant extract according to an embodiment of the present invention.
FIG. 4 is a graph showing the results of confirming toxicity of Sosa tree and its plant extracts on cells according to an embodiment of the present invention.
FIG. 5 is a graph showing the results of examining the toxicity of Wang cherry tree and its plant extracts to cells according to an embodiment of the present invention.
FIG. 6 is a graph showing the results of confirming the osteoclast differentiation inhibitory effect of the rock horse chestnut and its inborn plant extract according to an embodiment of the present invention.
FIG. 7 is a graph showing the results of confirming the osteoclast differentiation inhibitory effect of the rock horse chestnut and its inborn plant extract according to an embodiment of the present invention.
FIG. 8 is a graph showing the results of confirming the osteoclast differentiation inhibition effect of the extract of the nursery tree and the herbaceous plant according to an embodiment of the present invention.
FIG. 9 is a graph showing the results of confirming the osteoclast differentiation inhibitory effect of a nursery tree and its plant extracts according to an embodiment of the present invention.
FIG. 10 is a graph showing the results of confirming the osteoclast differentiation inhibitory effect of canola and its plant extracts according to an embodiment of the present invention.
FIG. 11 is a graph showing the results of confirming the osteoclast differentiation inhibitory effect of canola and its plant extracts according to an embodiment of the present invention.
12 is a graph showing the results of confirming the osteoclast differentiation inhibitory effect of Sosa tree and its inborn plant extract according to an embodiment of the present invention.
FIG. 13 is a graph showing the results of confirming the osteoclast differentiation inhibitory effect of Sosa tree and its inborn plant extract according to an embodiment of the present invention.
FIG. 14 is a graph showing the results of confirming the osteoclast differentiation inhibitory effect of the Yoshino cherry tree and its plant extracts according to an embodiment of the present invention.
FIG. 15 is a graph showing the results of confirming the osteoclast differentiation inhibitory effect of the Yoshino cherry tree and its in vivo plant extract according to an embodiment of the present invention.

Hereinafter, the present invention will be described in more detail with reference to Examples. It will be apparent to those skilled in the art that these embodiments are only for describing the present invention in more detail and that the scope of the present invention is not limited by these embodiments in accordance with the gist of the present invention .

[Example]

[Manufacturing Example] Materials used for extraction and extraction conditions

In order to examine the effect of the extract according to the present invention on preventing or treating bone diseases, 33 kinds of plants were extracted under extraction condition 1 or 2. The types of plants and extraction conditions are shown in Tables 1 and 2 below.

number Botanical name Extraction site Scientific name extraction
Condition 7-1 Leather tree leaf Ailanthus altissima (Mill.) Swingle for. Altissima 2 7-4 Geese strawberry leaf There is Rubus longisepalus. dustawai (Nakai) T.B.Lee 2 7-5 Bark tree leaf Platycarya strobilacea Siebold & Zucc. there is. strobilacea for. strobilacea 2 7-6 Goldmatt Above ground Patrinia saniculaefolia Hemsl. 2 7-7 Cold Above ground + flower Capsella bursapastoris (L.) L.W.Medicus 2 7-8 Ivy vine Leaf + stem Parthenocissus tricuspidata (Siebold & Zucc.) Planch. 2 7-9 Horseshoe Leaf + stem Thlaspi arvense L. One 7-10 Rock spruce tree Leaf + flower There is Deutzia grandiflora. baroniana Diels 2 7-11 Flea spot Leaf + stem + seed Arenaria serpyllifolia L. 2 7-12 Bogor Root Psoralea corylifolia L. One 7-13 barley Flower I have Hordeum vulgare. hexastichon (L.) Asch. 2 7-14 barley Root I have Hordeum vulgare. hexastichon (L.) Asch. 2 7-15 barley Leaf + stem I have Hordeum vulgare. hexastichon (L.) Asch. 2 7-16 mulberry tree stem Morus alba L. One 7-17 Fir tree leaf Eurya japonica Thunb. 2 7-18 A mountain tree leaf Cornus kousa F.Buerger ex Miquel 2 7-19 Mountain leek Above ground Allium thunbergii G.Don One 7-20 Sanjo Pop Tree leaf Spiraea blumei G.Don 2 7-21 Triple seal Above ground Bupleurum falcatum L. One 7-22 Shout Fruit Rumex cripus L 2 7-23 Sosa tree leaf Carpinus turczaninovii Hance 2 7-25 celandine Leaf + stem I have Chelidonium majus. asiaticum (Hara) Ohwi One 7-26 Virtue tree leaf Mallotus japonicus (Thunb.) Muell. Arg. 2 7-27 Wando thorn tree leaf Ilex x wandoensis C.F.Miller 2 7-28 Dog wrestling leaf Boehmeria platanifolia Franch. et Sav 2 7-29 Yoshino cherry tree leaf Prunus yedoensis Matsum 2 7-30 The leaf Akebia quinata (Thunb.) Decne. 2 7-31 Overhead leaf Phytolacca esculenta VanHoutte 2 7-32 Chestnut leaf I have prunus serrulata. Pubescens (Makino) Nakai 2 7-33 Poppy tree Flower Spiraea prunifolia for. simpliciflora Nakai One

discrimination Extraction condition One Each sample powder was extracted with methanol (MethOH) using an Accelerated Solution Extractor (Dinex, USA) at 50 ° C, filtered and concentrated 2 Each sample powder was extracted with ethanol (EtOH) using an Accelerated Solution Extractor (Dinex, USA) at 50 ° C, filtered and concentrated.

[Comparative Example] Plants for comparison belonging to the plant and the copper according to the present invention

In order to compare the efficacy of the extract according to the present invention with that of the plants belonging to the genus Dongkuk, We purchased plants belonging to the family of the Yoshino cherry tree. The genera of the plants are shown in Table 3 below.

Invention of the present invention Inbred plant Scientific name Rock spruce tree
Deutzia prunifolia Plum blossoms Deutzia coreana Geography Deutzia coreana var. triradiata Water bounty Deutzia glabrata A lot of chaos Deutzia crenata for. plena A mountain tree
Cornus kousa Corn oil Cornus officinalis Dogwood Cornus controversa Dogwood Cornus walteri White wood Cornus alba Bear Cornus macrophylla Dog wrestling
Boehmeria platanifolia Moschino Boehmeria nivea Wang Moche Boehmeria pannosa Island Moschip Boehmeria nipononivea A little waxy tree Boehmeria spicata Turtle tail Boehmeria tricuspis Turtle tail There is Boehmeria tricuspis. unicuspis Sosa tree
Carpinus laxiflora Heaven tree Carpinus laxiflora Dogwood tree Carpinus tschonoskii Magpie Carpinus cordata Yoshino cherry tree
Prunus yedoensis Mountain cherry tree Prunus sargentii Dugong tree Prunus padus Cherry tree Prunus buergeriana cherry I have prunus serrulata. spontanea All cherry trees Prunus pendula for. ascendens Copy tree Prunus persica Island cherry tree Prunus takesimensis All cherry trees Prunus pendula for. ascendens Seoul Gulong I have a prunus padus. seoulensis Dog cherry tree Prunus leveilleana Seahorse Prunus mandshurica var. glabra Hairy dog I have Prunus leveilleana. pilosa Mountain copy Prunus davidiana Plum tree Prunus salicina Anchors Prunus tomentosa Apricot tree There is Prunus armeniaca. ansu A lady's neck I got a prunus salicina. columnalis

[Example 1] Isolation and culture of mouse bone marrow macrophages

In order to confirm the effect of preventing or treating bone diseases, the mouse bone marrow macrophages were isolated and cultured in the following manner for the extracts obtained in the above Preparation Examples and Comparative Examples.

Three-week-old male ICR mice (Nari Biotech, Korea) were sacrificed by cervical dislocation, and then the hindlimb outer skin of the rat was removed using a saddle seat. minimum essential medium alpha; Gibco, USA). In addition, the bones in the muscles were separated using tweezers, transferred to a new α-MEM, 600 μl of α-MEM was placed in a syringe and inserted into the central spinal part of the separated leg bone and injected 2 to 3 times Bone marrow cells were harvested. The extracted bone marrow cells were centrifuged to remove the supernatant, and then a fresh α-MEM was added thereto to separate the bone marrow macrophages from the bone marrow cells. A histopaque (Sigma -Aldrich, USA) was used to finally isolate the bone marrow macrophages.

The bone marrow macrophages isolated from the above procedure were treated with 1% antibiotic-antimycotic (Gibco, USA), 10% fetal bovine serum (FBS) (Gibco, USA) and M-CSF culture medium containing 30 ng / ml of macrophage-colony stimulating factor (R & D system Inc., USA) at 37 ° C in a 5% CO ₂ incubator.

[Example 2] Determine whether the extract is cytotoxic

In order to confirm the toxicity of the extracts of the preparation examples and the comparative examples according to the present invention to cells, a cytotoxicity experiment was conducted. The cell culture medium used was DMEM (Dulbecco's Modified Eagle's medium) supplemented with 1% antibiotic-antimicrobial solution and 10% FBS (fetal bovine serum). To each well of a 96-well plate, 1 x 10 ⁴ mouse macrophages were dispensed and cultured for 24 hours at 37, 5% CO ₂ . Then, each extract was added to 96-well plates in which mouse macrophages were dispensed, and further cultured for 24 hours. A 5mg / ml MTT (Thiazolyl Blue Tetrazolium Bromide) solution was added in each 20μl per well. After 24 hours, the reaction was carried out at 37, 5% CO ₂ cell incubator for 4 hours. After removing the medium, 100 μl of DMSO was added, dissolved in a shaker for 20 minutes, and the absorbance was measured at 570 nm using an ELISA reader to confirm cell viability and toxicity of the sample. Cell viability was calculated as a percentage of the survival rate of the experimental group (each well treated with each extract) versus the survival rate of the control (well treated with well). The results are shown in Figs. 1 to 5 and Tables 4 to 8 as percentages based on the survival rate of the control group.

As shown in Fig. 1, Table 4 and Table 5, it was found that only the plum blossom tree trunk extract showed low cell viability at the high concentration and the other plant extracts did not have cytotoxicity .

Botanical name The extract concentration (占 퐂 / ml) Absorbance value Cell survival rate (%) - - 1.059 + 0.117 100 A diver
Frequently
Leaves, flowers One 1.326 + - 0.252 125 3 1.333 + 0.129 126 5 1.309 + 0.050 124 10 1.547 + 0.147 146 20 1.768 + 0.064 167 40 2.075 + - 0.262 196 80 1.817 ± 0.296 172 A diver
Frequently
stem One 1.081 + - 0.105 102 3 1.056 + 0.082 100 5 1.093 + 0.077 103 10 1.094 + - 0.100 103 20 1.201 + 0.168 113 40 1.415 ± 0.191 134 80 1.640 + 0.125 155 Plum
Valdor
stem One 0.926 + 0.160 87 3 0.925 + 0.091 87 5 1.020 + - 0.233 96 10 1.228 + 0.166 116 20 1.175 + 0.097 111 40 0.834 + 0.088 79 80 0.309 + 0.084 29 Plum
Valdor
leaf One 0.965 + 0.016 91 3 1.111 ± 0.163 105 5 1.190 + - 0.052 112 10 1.545 + - 0.234 146 20 1.541 + 0.081 146 40 1.829 + 0.065 173 80 2.321 + 0.130 219 Water bounty
petal One 1.002 + 0.068 95 3 1.001 + 0.146 95 5 1.123 + 0.063 106 10 1.350 + 0.074 128 20 1.614 + 0.186 153 40 2.325 + 0.248 220 80 2.380 + - 0.211 225

Botanical name Extract concentration
(占 퐂 / ml) Absorbance value Cell survival rate (%) - - 1.102 + 0.156 100 Water bounty
stem




One 1.088 + 0.127 99 3 1.170 + 0.018 106 5 1.343 + 0.127 122 10 1.375 + 0.149 125 20 1.702 + 0.263 154 40 1.136 + 0.246 103 80 1.162 + - 0.01 105 Rock horse
Valdor
leaf



One 1.127 + 0.138 102 3 1.104 + 0.13 100 5 1.276 + - 0.102 116 10 1.276 + 0.036 116 20 1.631 + 0.131 148 40 1.592 + 0.133 144 80 1.975 + 0.143 179 Rock horse
Valdor
stem



One 0.953 + 0.208 86 3 1.276 + 0.224 116 5 1.265 ± 0.051 115 10 1.36 + 0.134 123 20 1.489 + 0.073 135 40 1.305 + 0.139 118 80 2.297 + 0.235 208 Geography
Leaves, stem




One 0.878 + 0.191 80 3 1.017 + 0.226 92 5 1.057 + - 0.153 96 10 1.229 + 0.111 112 20 1.525 + 0.177 138 40 1.523 + - 0.218 138 80 2.424 + 0.211 220

As shown in FIG. 2, Table 6, and Table 7, among the nursery tree and its plants, a high concentration treatment was performed on the leaf and stem extracts of the seedlings, the leafy and stem extracts of the woody trees, Cell survival rate.

Botanical name Extract concentration
(占 퐂 / ml) Absorbance value Cell survival rate (%) - - 0.912 + 0.064 100 Bear
leaf




One 0.926 + 0.038 102 3 1.168 ± 0.058 128 5 1.15 + 0.024 126 10 1.809 + 0.116 198 20 2.251 + - 0.113 247 40 2.266 + 0.086 248 80 1.236 + 0.119 135 Bear
Stem-bark




One 1.05 + 0.166 115 3 1.172 + 0.038 128 5 1.353 + - 0.054 148 10 1.82 + 0.118 199 20 2.423 + 0.073 266 40 2.112 + 0.097 231 80 1.075 + 0.095 118 Dogwood
Leaves, stem




One 0.963 + 0.093 106 3 1.023 0.011 121 5 1.165 + 0.115 134 10 1.187 ± 0.058 130 20 1.473 + 0.176 161 40 1.466 + 0.033 161 80 0.551 + 0.04 60 Dogwood
Stem-bark




One 0.957 + 0.096 105 3 1.205 + 0.112 132 5 1.487 + 0.095 163 10 2.01 + 0.094 220 20 2.438 + 0.17 267 40 1.926 + 0.122 211 80 0.708 + 0.169 78 Dogwood
Leaves, stem




One 0.961 + 0.079 105 3 1.03 + - 0.112 113 5 1.269 + 0.033 139 10 1.688 + 0.144 185 20 2.119 + 0.065 232 40 2.115 + 0.133 232 80 0.774 + 0.076 85 Botanical name Extract concentration
(占 퐂 / ml) Absorbance value Cell survival rate (%) - - 0.793 + - 0.059 100 Dogwood
Disease




One 0.89 + 0.094 112 3 1.307 + 0.023 165 5 1.495 + 0.087 189 10 2.018 ± 0.127 255 20 2.569 + 0.124 324 40 0.947 + 0.124 119 80 0.379 ± 0.056 48 Dogwood
Fruit




One 0.991 + 0.147 125 3 0.916 ± 0.001 116 5 1.026 + 0.117 129 10 1.067 + 0.067 135 20 1.105 ± 0.05 139 40 1.13 + 0.076 143 80 1.472 + 0.133 186 White wood
Leaves, stem




One 1.027 + 0.065 130 3 1.141 ± 0.05 144 5 1.121 + - 0.121 141 10 1.249 + 0.011 158 20 1.978 + - 0.12 250 40 1.229 + 0.151 155 80 0.457 + 0.109 58 A mountain tree
stem




One 0.86 + 0.019 108 3 0.873 + 0.083 110 5 0.991 + 0.097 125 10 0.993 + 0.109 125 20 1.283 + 0.137 162 40 1.807 + 0.061 228 80 1.715 + 0.108 216 A mountain tree
Fruit




One 0.843 + 0.039 106 3 0.885 + 0.042 112 5 0.915 + 0.078 115 10 0.955 + 0.113 120 20 1.014 + 0.043 128 40 1.021 + 0.041 129 80 0.97 + 0.068 122

Botanical name Extract concentration
(占 퐂 / ml) Absorbance value Cell survival rate (%) - - 0.836 ± 0.05 100 A mountain tree
leaf




One 0.842 + 0.124 101 3 1.069 + 0.085 128 5 1.01 ± 0.119 121 10 1.378 + 0.093 165 20 1.469 + 0.092 176 40 1.213 + 0.124 145 80 0.943 + 0.108 113 A mountain tree
Stem-bark




One 0.8 ± 0.115 96 3 0.88 + 0.033 105 5 0.904 + 0.128 108 10 1.068 + 0.084 128 20 1.059 + 0.109 127 40 1.481 + 0.082 177 80 1.938 + 0.135 232 Corn oil
Leaves, stem




One 0.778 + 0.024 93 3 0.961 + 0.089 115 5 1.156 + 0.048 138 10 1.575 + 0.119 188 20 1.607 + 0.126 192 40 1.435 + 0.093 172 80 1.395 + 0.13 167 Corn oil
Fruit




One 0.788 + 0.061 94 3 1.018 + 0.119 122 5 1.124 + 0.048 135 10 1.14 ± 0.093 136 20 1.399 + 0.088 167 40 1.502 + 0.082 180 80 0.426 + 0.036 51

As shown in FIG. 3 and Table 8, it was confirmed that low cell survival rate was observed in the high concentration treatment of the outbred extract of the canola and the turtle tail outpost extract among the canine and its copepods. It was confirmed that Wang Mousypole exhibited cytotoxicity at lower concentrations than other plant extracts.

Botanical name Extract concentration
(占 퐂 / ml) Absorbance value Cell survival rate (%) - - 1.43 + 0.031 100 Moschino
outpost


One 0.835 + 0.031 58 5 1.249 + 0.137 87 10 1.126 + 0.091 79 50 1.849 + 0.134 129 100 2.171 ± 0.2 152 Wang Moche
outpost


0.01 1.419 + - 0.057 99 0.05 1.432 + 0.032 100 0.1 1.332 ± 0.007 93 0.5 1.179 + 0.133 82 One 0.983 + 0.122 69 Island Moschip
Above ground


One 1.832 + 0.026 128 5 1.692 + 0.025 118 10 1.713 + - 0.1 120 50 1.46 ± 0.101 102 100 1.2 ± 0.183 84 Dog wrestling
outpost


One 1.691 + 0.086 118 5 1.674 + 0.094 117 10 1.55 + 0.082 108 50 0.19 + 0.055 13 100 0.209 + 0.045 15 Little bitch
tree
outpost

One 1.455 + 0.0815 102 5 1.482 + 0.2888 104 10 1.933 + 0.359 135 50 2.483 + 0.2737 174 100 2.24 + 0.3269 157 Turtle tail
outpost


One 1.895 + 0.092 132 5 1.809 + 0.189 126 10 1.803 + 0.172 126 50 0.271 + 0.071 19 100 0.27 + 0.076 19 Turtle tail
outpost


One 1.374 + 0.123 96 5 1.276 + 0.259 89 10 1.19 0.355 83 50 1.001 ± 0.08 70 100 0.712 + - 0.104 50

As shown in Fig. 4, Table 9 and Table 10, there was no cytotoxicity in the extracts of black currant leaf and cercus leaf and in the other extracts, some cytotoxicity was observed in the high concentration treatment Respectively.

Botanical name Extract concentration
(占 퐂 / ml) Absorbance value Cell survival rate (%) - - 1.403 + - 0.185 100 Dogwood tree
Stem-bark




One 1.414 + 0.078 101 3 1.279 + 0.034 91 5 1.401 + - 0.1 100 10 1.638 + 0.149 117 20 0.99 ± 0.167 71 40 0.136 + 0.034 10 80 0.165 ± 0.007 12 Dogwood tree
Leaves, stem




One 1.336 + - 0.104 95 3 1.49 + - 0.054 106 5 1.585 + 0.124 113 10 1.316 + 0.231 94 20 0.815 + 0.031 58 40 0.29 + 0.082 21 80 0.288 ± 0.059 20 Magpie
Branch




One 1.22 + 0.079 87 3 1.214 + 0.213 86 5 1.223 + - 0.211 87 10 1.424 + 0.173 101 20 1.584 + 0.125 113 40 0.217 + - 0.107 15 80 0.231 + 0.033 16 Magpie
leaf




One 1.037 + 0.067 74 3 1.099 + 0.043 78 5 1.167 + 0.066 83 10 1.163 + 0.075 83 20 1.431 + - 0.286 102 40 1.649 + 0.176 118 80 1.455 + 0.046 104

Botanical name Extract concentration
(占 퐂 / ml) Absorbance value Cell survival rate (%) - - 0.95 + 0.064 100 Heaven tree
Flower




One 1.359 + 0.022 143 3 1.43 + 0.218 150 5 1.241 + 0.228 131 10 1.417 + 0.17 149 20 1.526 + 0.17 161 40 0.868 + 0.117 91 80 0.175 + 0.048 18 Heaven tree
leaf




One 1.304 + 0.163 137 3 1.366 + 0.129 144 5 1.16 ± 0.066 122 10 1.187 + 0.196 125 20 1.219 ± 0.2 128 40 1.336 + 0.132 141 80 0.589 + 0.136 62 Heaven tree
Stem-bark




One 0.91 + 0.146 96 3 0.98 + 0.085 103 5 1.13 + - 0.105 119 10 1.359 + 0.07 143 20 1.595 + 0.095 168 40 0.147 ± 0.007 16 80 0.126 0.003 13 Sosa tree
Leaves, stem




One 1.218 + 0.144 128 3 1.322 + 0.121 139 5 1.318 + 0.07 139 10 0.526 + 0.074 55 20 0.154 + - 0.01 16 40 0.2 ± 0.026 21 80 0.196 + 0.023 21

As shown in Fig. 5 and Tables 11 to 15, it was confirmed that both the Yoshino cherry and its plants were not cytotoxic.

Botanical name Extract concentration
(占 퐂 / ml) Absorbance value Cell survival rate (%) - - 0.842 + 0.039 100 Dog cherry tree
Stem-bark




One 0.902 + 0.055 107 3 1.11 + 0.066 132 5 1.119 + 0.036 133 10 1.453 + - 0.059 173 20 1.741 + 0.067 207 40 2.004 ± 0.141 238 80 2.219 + 0.127 264 Seahorse
leaf




One 0.874 + 0.043 104 3 0.87 ± 0.108 103 5 0.874 + 0.123 104 10 0.905 + 0.107 108 20 0.996 + 0.085 118 40 1.143 + - 0.102 136 80 2.05 + 0.064 244 Seahorse
stem




One 0.725 + 0.072 86 3 0.845 + 0.067 100 5 0.932 + 0.116 111 10 1.14 ± 0.037 135 20 1.142 + 0.078 136 40 1.764 + 0.077 210 80 2.165 ± 0.106 257 Seahorse
Stem-bark




One 0.803 + 0.069 95 3 0.842 + 0.039 100 5 0.932 + 0.033 111 10 1.248 + 0.031 148 20 1.47 ± 0.003 175 40 1.788 + 0.143 212 80 1.901 + 0.098 226 Dugong tree
Leaf, stem, flower




One 0.804 + 0.076 96 3 0.881 + 0.037 105 5 0.878 ± 0.053 104 10 1 ± 0.133 119 20 1.068 + 0.096 127 40 1.564 ± 0.059 186 80 1.957 + 0.163 232

Botanical name Extract concentration
(占 퐂 / ml) Absorbance value Cell survival rate (%) - - 0.734 + 0.094 100 cherry
leaf




One 0.56 + 0.034 76 3 0.575 ± 0.059 78 5 0.767 ± 0.059 104 10 1.044 + 0.075 142 20 1.149 + 0.098 157 40 1.631 + 0.096 222 80 1.996 + 0.136 272 cherry
Stem-bark




One 0.812 + 0.048 111 3 0.936 + 0.13 127 5 0.987 + - 0.102 134 10 1.458 ± 0.122 199 20 1.511 + - 0.051 206 40 1.63 + - 0.108 222 80 1.808 0.11 246 Copy tree
Flower




One 0.729 + 0.109 99 3 0.819 + 0.036 112 5 0.77 + 0.055 105 10 0.802 + 0.137 109 20 0.792 + 0.04 108 40 1.144 + 0.115 156 80 1.678 ± 0.073 228 Copy tree
Stem-bark




One 1.003 ± 0.132 137 3 1.18 ± 0.113 161 5 1.425 + 0.114 194 10 1.755 + - 0.12 239 20 2.062 + 0.084 281 40 2.208 + 0.116 301 80 2.218 + 0.117 302 Copy tree
Leaves, stem




One 0.922 + 0.135 126 3 0.966 + 0.093 131 5 1.082 + 0.101 147 10 1.121 + 0.036 153 20 1.325 + 0.106 180 40 2.113 + 0.091 288 80 2.139 + 0.038 291

Botanical name Extract concentration
(占 퐂 / ml) Absorbance value Cell survival rate (%) - - 0.713 ± 0.05 100 Mountain cherry tree
leaf




One 0.73 + 0.045 102 3 0.876 + 0.017 123 5 1.058 + 0.088 148 10 1.194 + 0.105 167 20 1.329 + 0.03 186 40 2.036 + 0.04 285 80 2.181 + - 0.111 306 Mountain cherry tree
stem




One 0.866 ± 0.051 121 3 0.888 + 0.103 124 5 1.133 + 0.06 159 10 1.345 + 0.067 189 20 1.532 + 0.098 215 40 2.083 + 0.067 292 80 2.219 ± 0.058 311 Apricot tree
leaf




One 0.823 + 0.124 115 3 0.873 + 0.152 122 5 0.915 + 0.047 128 10 0.891 + - 0.13 125 20 1.078 + - 0.1 151 40 1.345 ± 0.008 189 80 1.467 0.041 206 Apricot tree
stem




One 0.876 + 0.028 123 3 0.997 + 0.022 140 5 1.034 + 0.092 145 10 1.107 + 0.098 155 20 1.147 + 0.133 161 40 1.315 + 0.047 184 80 1.666 + 0.072 234 Apricot tree
Stem-bark




One 0.926 + 0.023 130 3 0.855 + 0.072 120 5 0.959 + 0.092 134 10 1.181 + 0.044 166 20 1.211 + - 0.13 170 40 2.238 + 0.119 314 80 2.332 + 0.081 327

Botanical name Extract concentration
(占 퐂 / ml) Absorbance value Cell survival rate (%) - - 1.203 + 0.089 100 Cherry tree
leaf




One 1.348 + 0.195 112 3 1.369 + 0.197 114 5 1.548 + 0.123 129 10 1.628 + 0.019 135 20 1.69 ± 0.159 141 40 1.841 + - 0.104 153 80 2.331 + 0.201 194 Cherry tree
Stem-bark




One 1.209 + 0.136 101 3 1.478 + - 0.232 123 5 1.627 + 0.127 135 10 1.769 ± 0.189 147 20 1.578 + 0.061 131 40 2.392 + 0.032 199 80 3.04 0.078 253 Island cherry tree
leaf




One 1.166 + 0.175 97 3 1.476 + 0.073 123 5 1.478 + 0.147 123 10 1.671 + 0.074 139 20 1.546 + 0.072 129 40 1.936 + 0.091 161 80 2.381 + 0.171 198 Island cherry tree
Stem-bark




One 1.221 + 0.129 101 3 1.651 + 0.194 137 5 1.537 + 0.134 128 10 1.941 + 0.115 161 20 1.983 + - 0.231 165 40 3.026 + 0.089 252 80 3.382 + 0.159 281 Anchors
Leaves, stem




One 1.123 + - 0.12 93 3 1.157 + - 0.102 96 5 1.225 + - 0.122 102 10 1.133 + 0.029 94 20 1.142 + 0.074 95 40 1.666 + 0.096 138 80 1.676 ± 0.052 139

Botanical name Extract concentration
(占 퐂 / ml) Absorbance value Cell survival rate (%) - - 0.921 + - 0.052 100 A lady's neck
Leaves, stem




One 1.092 + 0.065 118 3 1.02 + - 0.155 111 5 1.194 + 0.116 130 10 1.478 + 0.118 160 20 1.774 + 0.087 193 40 2.346 + 0.171 255 80 2.405 + 0.139 261 All cherry trees
Stem-bark




One 1.099 + 0.121 119 3 1.198 + 0.126 130 5 1.132 ± 0.119 123 10 1.804 + 0.215 196 20 2.156 + 0.355 234 40 3.029 ± 0.05 329 80 3.104 + - 0.052 337 All cherry trees
Leaves, stem




One 1.066 + 0.098 116 3 1.097 + 0.065 119 5 1.063 + 0.076 115 10 1.635 + 0.169 177 20 2.079 ± 0.02 226 40 2.109 ± 0.107 229 80 2.34 + 0.128 254 Yoshino cherry tree
Leaves, flowers




One 1.054 + 0.036 114 3 0.962 + 0.132 104 5 0.964 + 0.116 105 10 1.266 + 0.033 137 20 1.261 + 0.042 137 40 1.918 + 0.129 208 80 2 ± 0.033 217 Yoshino cherry tree
stem




One 1.091 + 0.074 118 3 1.283 + 0.145 139 5 1.232 ± 0.009 134 10 2.166 + 0.055 235 20 2.285 + 0.089 248 40 2.824 ± 0.089 307 80 3.089 + 0.097 335 Plum tree
stem




One 0.982 + 0.182 105 3 1.276 + 0.123 137 5 1.232 + 0.116 132 10 1.28 + - 0.14 139 20 1.616 + 0.157 175 40 2.202 + 0.18 239 80 2.49 + 0.123 270

[Example 3] Identification of inhibition of differentiation into osteoclasts

In order to confirm that the differentiation of bone marrow macrophages into osteoclasts induced by RANKL (receptor activator of nuclear factor kappa-B ligand) is inhibited by the extract according to the present invention, experiments for confirming inhibition of osteoclast differentiation were performed &Quot; Park EK. Et al., Biochem Biophys Res Commun., 325 (4): 1472-1480 (2004)).

For confirmation of differentiation inhibition rate, the bone marrow macrophages prepared by the method of Example 1 were dispensed into 96-wells at 5 × 10 ⁴ cells / well. Each extract was then mixed with α-MEM per concentration (1% antibiotic-antimicrobial solution, 10% FBS, 30 ng / ml M-CSF and 50 ng / ml RANKL) to a final volume of 200 μl After the preparation, the cells were treated with the above-mentioned mixed bone marrow macrophages and replaced with fresh medium every 2 days in a cell incubator under 37, 5% CO ₂ condition, and finally, the cells were cultured for 5 days to prepare an experimental group. As a negative control, cells were prepared using α-MEM supplemented with 1% antibiotic-antimicrobial solution, 10% FBS, and 30 ng / ml of M-CSF. As a positive control, 1% antibiotic- Solution, 10% FBS, 30 ng / ml M-CSF, and α-MEM supplemented with 50 ng / ml RANKL. The number of osteoclasts with three or more nuclei was measured by light microscope after TRAP assay (Sigma-Aldrich, USA) with TRAP assay kit (Sigma-Aldrich, USA) 6 to 15 and Table 16 to Table 21.

Botanical name Extract concentration
(占 퐂 / ml) Number of RANKL-induced osteoclasts Osteoclast
Differentiation inhibition rate (%) 0 ± 0 RANKL 247 ± 8 0 A lot of chaos
Leaves, flowers
RANKL + 10 239 ± 1 3 RANKL + 20 244 ± 6 One RANKL + 40 102 ± 8 59 A lot of chaos
stem
RANKL + 10 218 ± 3 12 RANKL + 20 193 ± 5 22 RANKL + 40 164 ± 6 33 Plum blossoms
stem
RANKL + 10 209 ± 1 15 RANKL + 20 176 ± 5 29 RANKL + 40 157 ± 1 37 Plum blossoms
leaf
RANKL + 10 192 ± 6 22 RANKL + 20 118 ± 3 52 RANKL + 40 0 ± 0 100 Water bounty
petal
RANKL + 10 227 ± 4 8 RANKL + 20 212 ± 4 14 RANKL + 40 167 ± 2 32 Water bounty
stem
RANKL + 10 221 ± 4 10 RANKL + 20 166 ± 6 33 RANKL + 40 147 ± 4 40 Rock spade
leaf
RANKL + 10 208 ± 4 16 RANKL + 20 177 ± 4 28 RANKL + 40 49 ± 1 80 Rock spade
stem
RANKL + 10 245 ± 8 One RANKL + 20 218 ± 2 12 RANKL + 40 169 ± 10 31 Geography
Leaves, stem
RANKL + 10 249 ± 1 0 RANKL + 20 203 ± 4 18 RANKL + 40 147 ± 4 40

As shown in FIGS. 8, 9 and 17, in the case of bear plants, the extracts of leaves and stems (bark) showed high inhibition rates of osteoclast differentiation, Leaf and stem extracts of leaves and stem, and stem and bark extracts of leaves and stem extracts of the stratum corneum showed high osteoclast differentiation inhibition rates. However, it was confirmed that the rate of osteoclast differentiation was slightly lower in the fruit of the stratum. In addition, it was confirmed that the extracts of stem and stem extracts, stem extracts, stem extracts and stem (bark) extracts of white mulberry trees exhibited a significantly high inhibition rate of differentiation, while those of the fruit extracts of mulberry trees showed a low inhibition rate of osteoclast differentiation Through this, it was confirmed that even the same plants exhibit different effects depending on tissues such as leaves and stems. In addition, it was confirmed that osteoclast differentiation inhibition rates were also high in leaves and stem extracts and fruit extracts of corn oil.

Botanical name Extract concentration
(占 퐂 / ml) Number of RANKL-induced osteoclasts Inhibition rate of differentiation into osteoclast (%) - - 0 ± 0 - - RANKL 252 ± 4 0 Bear
leaf
RANKL + 10 9 ± 1 97 RANKL + 20 0 ± 0 100 RANKL + 40 0 ± 0 100 Bear
Stem-bark
RANKL + 10 0 ± 0 100 RANKL + 20 0 ± 0 100 RANKL + 40 0 ± 0 100 Dogwood
Leaves, stem
RANKL + 10 138 ± 11 45 RANKL + 20 7 ± 2 97 RANKL + 40 0 ± 0 100 Dogwood
Stem-bark
RANKL + 10 0 ± 0 100 RANKL + 20 0 ± 0 100 RANKL + 40 0 ± 0 100 Dogwood
Leaves, stem
RANKL + 10 69 ± 2 73 RANKL + 20 16 ± 3 94 RANKL + 40 0 ± 0 100 Dogwood
Disease
RANKL + 10 24 ± 1 90 RANKL + 20 0 ± 0 100 RANKL + 40 0 ± 0 100 Dogwood
Fruit
RANKL + 10 259 ± 12 0 RANKL + 20 185 ± 12 27 RANKL + 40 113 ± 7 55 White wood
Leaves, stem
RANKL + 10 33 ± 1 87 RANKL + 20 0 ± 0 100 RANKL + 40 0 ± 0 100 A mountain tree
stem
RANKL + 10 247 ± 16 2 RANKL + 20 189 ± 13 25 RANKL + 40 14 ± 1 95 A mountain tree
Fruit
RANKL + 10 254 ± 8 0 RANKL + 20 239 ± 5 5 RANKL + 40 207 ± 4 18 A mountain tree
leaf
RANKL + 10 256 ± 11 0 RANKL + 20 68 ± 4 73 RANKL + 40 0 ± 0 100 A mountain tree
Stem-bark
RANKL + 10 202 ± 23 20 RANKL + 20 56 ± 11 78 RANKL + 40 0 ± 0 100 Corn oil
Leaves, stem
RANKL + 10 242 ± 22 4 RANKL + 20 99 ± 4 61 RANKL + 40 0 ± 0 100 Corn oil
Fruit
RANKL + 10 247 ± 4 2 RANKL + 20 129 ± 1 49 RANKL + 40 0 ± 0 100

As shown in FIGS. 10, 11 and 18, it was confirmed that the outgrowth of the extracts of T. ganoderma and T. turtle tail outbreak extracts showed a high inhibitory rate of osteoclast differentiation among the germinated ginseng and its plants.

Botanical name Extract concentration
(占 퐂 / ml) Number of RANKL-induced osteoclasts
Osteoclast
Differentiation inhibition rate (%) - - 0 ± 0 - - RANKL 199 ± 2 0 Moschino
outpost

RANKL + 5 197 ± 11 One RANKL + 10 197 ± 5 One RANKL + 20 134 ± 10 32 RANKL + 50 98 ± 10 51 Wang Moche
outpost
RANKL + 0.1 187 ± 14 6 RANKL + 0.5 178 ± 9 11 RANKL + 1 174 ± 18 13 Island Moschip
Above ground

RANKL + 5 179 ± 10 10 RANKL + 10 179 ± 10 10 RANKL + 20 151 ± 15 24 RANKL + 50 145 ± 15 27 Dog wrestling
outpost

RANKL + 5 196 ± 10 One RANKL + 10 164 ± 10 17 RANKL + 20 133 ± 23 33 RANKL + 50 0 ± 0 100 Mulberry leaves
outpost

RANKL + 5 185 ± 16 7 RANKL + 10 173 ± 13 13 RANKL + 20 148 ± 4 25 RANKL + 50 118 ± 9 41 Turtle tail
outpost

RANKL + 5 146 ± 2 27 RANKL + 10 139 ± 7 30 RANKL + 20 0 ± 0 100 RANKL + 50 0 ± 0 100 Turtle tail
outpost

RANKL + 5 178 ± 3 10 RANKL + 10 164 ± 5 18 RANKL + 20 159 ± 13 20 RANKL + 50 131 ± 1 34

As shown in FIGS. 12, 13, and 19, among the Sosa tree and its plants, the stem (bark) extract, leaf and stem extracts of the Japanese white cedar trees, flower extracts of the cedar trees, And the inhibition rate of osteoclast differentiation was high in the extracts.

Botanical name Extract concentration
(占 퐂 / ml) RANKL-induced
Number of osteoclasts Inhibition rate of differentiation into osteoclast (%) - - 0 ± 0 - - RANKL 267 ± 4 0 Dogwood tree
Stem-bark
RANKL + 5 255 ± 6 4 RANKL + 10 88 ± 11 67 RANKL + 20 42 ± 8 84 Dogwood tree
Leaves, stem
RANKL + 5 244 ± 8 9 RANKL + 10 87 ± 4 68 RANKL + 20 0 ± 0 100 Magpie
Branch
RANKL + 5 169 ± 1 37 RANKL + 10 170 ± 5 37 RANKL + 20 103 ± 8 62 Magpie
leaf
RANKL + 5 239 ± 1 10 RANKL + 10 223 ± 4 16 RANKL + 20 149 ± 2 44 Heaven tree
Flower
RANKL + 10 233 ± 6 13 RANKL + 20 157 ± 5 41 RANKL + 40 27 ± 4 90 Heaven tree
leaf
RANKL + 10 222 ± 8 17 RANKL + 20 152 ± 11 43 RANKL + 40 74 ± 3 72 Heaven tree
Stem-bark
RANKL + 10 151 ± 4 43 RANKL + 20 50 ± 3 81 RANKL + 40 0 ± 0 100 Sosa tree
Leaves, stem
RANKL + 1 258 ± 2 4 RANKL + 3 245 ± 10 8 RANKL + 5 237 ± 12 11

As shown in Figs. 14, 15, 20, and 21, among the Yoshino cherry tree and its plants, leaf extract and stem extract, stem (bark) extract and cherry leaf extract, Extracts, Leaf Extracts and Stem Extracts from Mountain Cherry Blossoms, Apricot Tree Leaf Extracts, Stem Extracts from Chrysanthemum cherry Extract, Stem Extracts from Ishihara Cherry Blossom, Stem Extracts from Oryzant Indicating that the osteoclast differentiation inhibition rate is high.

Botanical name Extract concentration
(占 퐂 / ml) RANKL-induced
Number of osteoclasts Inhibition rate of differentiation into osteoclast (%) - - 0 ± 0 - - RANKL 269 ± 10 0 Dog cherry tree
Stem-bark
RANKL + 10 244 ± 5 9 RANKL + 20 208 ± 10 23 RANKL + 40 108 ± 6 60 Seahorse
leaf
RANKL + 10 193 ± 16 28 RANKL + 20 129 ± 21 52 RANKL + 40 13 ± 2 95 Seahorse
stem
RANKL + 10 169 ± 1 37 RANKL + 20 46 ± 8 83 RANKL + 40 14 ± 3 95 Seahorse
Stem-bark
RANKL + 10 221 ± 4 18 RANKL + 20 137 ± 5 49 RANKL + 40 25 ± 2 91 Dugong tree
Leaf, stem, flower
RANKL + 10 237 ± 5 12 RANKL + 20 235 ± 9 13 RANKL + 40 218 ± 4 19 cherry
leaf
RANKL + 10 218 ± 9 19 RANKL + 20 94 ± 2 65 RANKL + 40 0 ± 0 100 cherry
Stem-bark
RANKL + 10 205 ± 8 24 RANKL + 20 200 ± 13 26 RANKL + 40 79 ± 9 71 Copy tree
Flower
RANKL + 10 246 ± 8 9 RANKL + 20 175 ± 14 35 RANKL + 40 122 ± 2 55 Copy tree
Stem-bark
RANKL + 10 272 ± 8 0 RANKL + 20 156 ± 19 42 RANKL + 40 20 ± 1 93 Botanical name Extract concentration
(占 퐂 / ml) Number of RANKL-induced osteoclasts Inhibition rate of differentiation into osteoclast (%) - - 0 ± 0 - -
Copy tree
Leaves, stem
RANKL 269 ± 10 0 RANKL + 10 221 ± 18 18 RANKL + 20 210 ± 16 22 RANKL + 40 99 ± 13 63 Mountain cherry tree
leaf
RANKL + 10 217 ± 13 20 RANKL + 20 184 ± 6 32 RANKL + 40 0 ± 0 100 Mountain cherry tree
stem
RANKL + 10 165 ± 5 39 RANKL + 20 33 ± 1 88 RANKL + 40 0 ± 0 100 Apricot tree
leaf
RANKL + 10 212 ± 12 21 RANKL + 20 109 ± 1 59 RANKL + 40 22 ± 3 92 Apricot tree
stem
RANKL + 10 207 ± 16 23 RANKL + 20 174 ± 8 35 RANKL + 40 127 ± 7 53 Apricot tree
Stem-bark
RANKL + 10 234 ± 6 13 RANKL + 20 154 ± 6 43 RANKL + 40 70 ± 4 74 Cherry tree
leaf
RANKL + 10 250 ± 3 7 RANKL + 20 174 ± 6 35 RANKL + 40 126 ± 8 53 Cherry tree
Stem-bark
RANKL + 10 297 ± 5 0 RANKL + 20 128 ± 15 53 RANKL + 40 0 ± 0 100 Island cherry tree
leaf
RANKL + 10 253 ± 13 6 RANKL + 20 219 ± 15 19 RANKL + 40 78 ± 6 71

Botanical name Extract concentration
(占 퐂 / ml) RANKL-induced
Number of osteoclasts Osteoclast
Differentiation inhibition rate (%) - - 0 ± 0 - - RANKL 269 ± 10 0 Island cherry tree
Stem-bark RANKL + 10 180 ± 13 33 RANKL + 20 159 ± 4 41 RANKL + 40 53 ± 4 80 Anchors
Leaves, stem RANKL + 10 189 ± 8 30 RANKL + 20 172 ± 12 36 RANKL + 40 106 ± 4 61 A lady's neck
Leaves, stem RANKL + 10 251 ± 6 7 RANKL + 20 271 ± 2 0 RANKL + 40 194 ± 8 28 All cherry trees
Stem-bark RANKL + 10 267 ± 4 One RANKL + 20 187 ± 9 31 RANKL + 40 48 ± 6 82 All cherry trees
Leaves, stem RANKL + 10 261 ± 15 3 RANKL + 20 218 ± 14 19 RANKL + 40 68 ± 4 75 Yoshino cherry tree
Leaves, flowers RANKL + 10 243 ± 9 10 RANKL + 20 194 ± 15 28 RANKL + 40 71 ± 1 74 Yoshino cherry tree
stem RANKL + 10 121 ± 6 55 RANKL + 20 59 ± 6 78 RANKL + 40 0 ± 0 100 Plum tree
stem RANKL + 10 271 ± 4 0 RANKL + 20 270 ± 13 0 RANKL + 40 278 ± 6 0

In addition, as shown in Table 22 and Table 23, although there was some difference in the degree of suppression of osteoclast differentiation and the concentration of extract compared to the positive control corresponding to 0, it was confirmed that osteoclast differentiation was suppressed in most experimental groups. In addition, each of the extracts was found to have no or less cytotoxic concentrations, such as the leaves of Corolla, Eucalyptus, Leucophora and Blossom, Ivyvine and Stem, Yoshino cherry, Sosa, Leaf and Leaf Especially osteoclast differentiation into osteoclasts.

number Botanical name Concentration of extract
(μg / ml) Differentiated into RANKL
Number of osteoclasts Inhibition rate of differentiation into osteoclast (%) Negative control group - 0 ± 0 - Positive control group RANKL 381 ± 5 0 7-28 Leaf of dogs RANKL + 5 373 ± 6 2 RANKL + 9 253 ± 6 34 Positive control group RANKL 298 ± 9 0 7-10 Rock spade
Leaves and flowers RANKL + 5 17 ± 2 94 RANKL + 10 0 ± 0 100 7-18 Corolla leaves RANKL + 5 10 ± 1 97 RANKL + 10 0 ± 0 100 7-23 Sosa leaf RANKL + 5 184 ± 8 38 RANKL + 10 51 ± 4 83 7-29 Yoshino cherry tree leaves RANKL + 5 158 ± 9 47 RANKL + 10 107 ± 2 64

number Extract concentration
(μg / ml) Inhibition rate of differentiation into osteoclast (%) 7-1 10 Is cytotoxic 20 Is cytotoxic 7-4 10 40 20 47.1 7-5 10 65.9 20 100 7-6 10 59.8 20 67.5 7-7 10 42.3 20 60.4 7-8 10 72.1 20 100 7-9 10 13 20 28.4 7-10 10 100 20 100 7-11 10 23.1 20 31.2 7-12 10 36.1 20 36.4 7-13 10 28.7 20 34.5 7-14 10 25.3 20 40.3 7-15 10 19 20 7.77 7-16 10 39.9 20 27 7-17 10 0 20 24.2 7-18 10 100 20 100 7-19 10 10.6 20 28.1 7-20 10 4.2 20 6 7-21 10 28.1 20 23.1 7-22 10 99.1 20 100 7-23 10 85 20 100 7-25 10 8.1 20 15 7-26 10 0 20 12.6 7-27 10 15 20 52.5 7-28 10 99.4 20 100 7-29 10 71.8 20 86 7-30 10 29 20 37 7-31 10 33 20 35 7-32 10 20 20 52 7-33 10 18.2 20 68.4

From the above results, it was confirmed that the extracts according to the present invention can effectively inhibit the differentiation from macrophages to osteoclasts with low side effects. Thus, the extracts according to the present invention can be effectively used for prevention and treatment of bone diseases It can be confirmed that it can be used.

Although the present invention has been described in detail with reference to the above results, it is to be understood that the scope of the present invention is not limited to the above embodiments and that various changes and modifications can be made without departing from the technical idea of the present invention. It will be obvious to those of ordinary skill in the art.

Claims (13)

(Rosales) consisting of a rock-sparrow, a plum horsetail, a geographical sparrow, a water sparrow, and a chubby sparrow (Deutzia) A pharmaceutical composition for preventing or treating bone diseases, comprising at least one plant extract selected from the group of plants as an active ingredient.
The method according to claim 1,
Wherein the extract is any one or more extracts selected from the group consisting of leaves, flowers, stems, bark, disease, fruit, and outpost. The method according to claim 1,
Wherein said bone disease is caused by bone resorption of osteoclasts. The method according to claim 1,
The bone disease may be selected from the group consisting of osteoporosis, osteogenesis, rickets, osteopenia, fibrotic bone disease, intractable bone disease, osteoarthritis, bone atrophy, paget's disease, periodontitis, rheumatoid arthritis, And bone damage caused by bone metastasis of cancer cells.
Wherein the plant extract comprises at least one plant extract selected from the group consisting of Rosales, Deutzia, consisting of a marbled bean curd, a plum horsetail, a geomancy balsam, a water bounty, and a multitude of bivalves.
6. The method of claim 5,
Wherein the extract is any one or more extracts selected from the group consisting of leaves, flowers, stems, bark, disease, fruit, and whole plants.
6. The method of claim 5,
Wherein said bone disease is caused by bone resorption of osteoclasts.
6. The method of claim 5,
The bone disease may be selected from the group consisting of osteoporosis, osteogenesis, rickets, osteopenia, fibrotic bone disease, intractable bone disease, osteoarthritis, bone atrophy, paget's disease, periodontitis, rheumatoid arthritis, And bone damage caused by bone metastasis of cancer cells.
A. obtaining and pulverizing at least one plant selected from the group of Rosales, Deutzia, consisting of rock spruce, plum horsetail, geese sparrow, water sparrow, and multitude sparrow; And
B. A method for preparing a pharmaceutical composition for preventing or treating bone diseases, comprising extracting the powder with an organic solvent in a solvent extraction apparatus and then filtering.
10. The method of claim 9,
Wherein the temperature of the solvent extraction apparatus is 30 ° C to 60 ° C.
10. The method of claim 9,
Wherein the extracting step is carried out with ethanol or methanol.
10. The method of claim 9,
Wherein the method further comprises the step of concentrating the filtered filtrate after step B.
10. The method of claim 9,
The above-mentioned bone diseases include bone diseases such as osteoporosis, osteomalacia, rickets, osteopenia, fibrotic bone disease, osteoarthritis, osteoarthritis, bone atrophy, paget's disease, periodontitis, rheumatoid arthritis, Metabolic bone disease, and bone damage caused by bone metastasis of cancer cells.

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