JP2019026640A - Muscle activator - Google Patents
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- JP2019026640A JP2019026640A JP2018139871A JP2018139871A JP2019026640A JP 2019026640 A JP2019026640 A JP 2019026640A JP 2018139871 A JP2018139871 A JP 2018139871A JP 2018139871 A JP2018139871 A JP 2018139871A JP 2019026640 A JP2019026640 A JP 2019026640A
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Abstract
Description
本発明は、ベルガモット、オオバコエンドロ、ニイタカヨモギ、ワダソウ、トゲバンレイシ、キリンケツ、ワニナシ、ベニバナ、トウモロコシ、ツルナ、ソバ、ウド及びキカラスウリからなる植物群から選択される1種又は2種以上の植物抽出物を含有する筋肉活性化剤に関するものである。 The present invention relates to one or more plant extracts selected from the group of plants consisting of bergamot, psyllium, sagebrush, scorpion, spruce, giraffe, crocodile, safflower, corn, tuna, buckwheat, udder, and scallop cucumber. It is related with the muscle activation agent containing a thing.
筋肉は、筋芽細胞が筋管細胞へ分化し、さらに筋管細胞が融合した筋線維から成り立っている。この筋芽細胞を賦活し細胞増殖を促進することにより筋肉増強又は筋肉減衰の予防に効果が期待できる。さらには、筋タンパク質の合成を促進するmTORシグナルの下流因子であるp70S6K、rpS6を活性化 (リン酸化)させることでも同様の効果が期待できる。このように、筋芽細胞の賦活又はp70S6K、rpS6の活性化は、ミオパチーなどの筋肉疾患、老化に伴うサルコペニア(加齢性減弱症)やロコモティブシンドローム(運動器症候群)、長期間のギプス固定や運動不足による廃用性萎縮の予防及び/又は治療、運動時又は運動後の筋タンパク合成低下抑制等に有用であると考えられる。さらに、皮膚の下には筋肉があり皮膚全体を支えているが、老化等により筋肉の機能が低下すると皮膚を支える力が弱まり、皮膚の弾力低下、たるみ及びしわの形成が誘導されると考えられていることから、筋芽細胞の賦活又はp70S6K、rpS6の活性化により皮膚改善作用も期待される。
平均寿命の延びや少子高齢化の進行に伴い、健康を維持しつつ長生きするために、優れた筋芽細胞賦活作用又はp70S6K活性化作用、rpS6活性化作用を有し、かつ安全性の高い機能性食品や化粧品素材等の開発が切望されている。
Muscles are composed of muscle fibers in which myoblasts are differentiated into myotube cells and myotube cells are fused. By activating these myoblasts and promoting cell proliferation, an effect can be expected in preventing muscle strengthening or muscle wasting. Furthermore, the same effect can be expected by activating (phosphorylating) p70S6K and rpS6, which are downstream factors of mTOR signal that promotes the synthesis of muscle protein. As described above, activation of myoblasts or activation of p70S6K and rpS6 is caused by muscular diseases such as myopathy, sarcopenia associated with aging, locomotive syndrome (motor organ syndrome), long-term cast fixation, It is considered useful for prevention and / or treatment of disuse atrophy due to lack of exercise, suppression of decrease in muscle protein synthesis during or after exercise. Furthermore, there are muscles under the skin that support the whole skin, but if the function of the muscles declines due to aging, etc., the ability to support the skin weakens, leading to reduced skin elasticity, sagging and wrinkle formation. Therefore, a skin improvement effect is also expected by activation of myoblasts or activation of p70S6K and rpS6.
Along with the prolongation of life expectancy and the declining birthrate and aging, it has excellent myoblast activation action, p70S6K activation action, rpS6 activation action and high safety function in order to live long while maintaining health Development of foodstuffs and cosmetic materials is eagerly desired.
本発明は、新規な筋肉活性化剤を提供することを主な目的とする。 The main object of the present invention is to provide a novel muscle activator.
本発明者は、上記課題を解決すべく検討した結果、ベルガモット、オオバコエンドロ、ニイタカヨモギ、ワダソウ、トゲバンレイシ、キリンケツ、ワニナシ、ベニバナ、トウモロコシ、ツルナ、ソバ、ウド及びキカラスウリからなる植物群から選択される1種又は2種以上の植物抽出物が優れた筋芽細胞賦活作用を有することを見出し、さらには、ベルガモット、オオバコエンドロ、ニイタカヨモギ、ワダソウ、キカラスウリにp70S6K活性化作用を見出し、本発明を完成した。 As a result of studying to solve the above problems, the present inventor has selected from a plant group consisting of bergamot, psyllium, sardine mugwort, waddows, spiders, giraffe, crocodile, safflower, corn, tuna, buckwheat, udo and kikarasuuri It is found that one or two or more kinds of plant extracts have an excellent myoblast activation action, and furthermore, a p70S6K activation action is found in bergamot, psyllium, niitaka mugwort, wadaso, kakirasuri, Completed the invention.
本発明として、下記のものを挙げることができる。
(1)ベルガモット、オオバコエンドロ、ニイタカヨモギ、ワダソウ、トゲバンレイシ、キリンケツ、ワニナシ、ベニバナ、トウモロコシ、ツルナ、ソバ、ウド及びキカラスウリからなる植物群から選択される1種又は2種以上の植物抽出物を有効成分として含有する、筋肉活性化剤。
(2)筋肉活性化剤が筋肉増強剤又は筋肉減衰予防剤である、前記(1)に記載の筋肉活性化剤。
(3)筋肉活性化剤が皮膚改善剤である、前記(1)に記載の筋肉活性化剤。
Examples of the present invention include the following.
(1) One or more plant extracts selected from the plant group consisting of bergamot, plantain, Japanese sagebrush, yellow-spotted swordfish, spruce, giraffe, crocodile, safflower, corn, tuna, buckwheat, udo A muscle activating agent containing as an active ingredient.
(2) The muscle activator according to (1), wherein the muscle activator is a muscle enhancer or a muscle attenuation preventive agent.
(3) The muscle activator according to (1), wherein the muscle activator is a skin improving agent.
本発明に係る筋肉活性化剤は、筋芽細胞の賦活又は筋タンパク質の合成を促進することにより、筋肉増強剤、筋肉減衰予防剤、皮膚改善剤としての効果が期待される。本発明にかかる筋肉活性化剤は、食用植物の成分由来であることから安全性が高いものである。 The muscle activator according to the present invention is expected to be effective as a muscle enhancer, a muscle attenuation preventing agent, and a skin improving agent by promoting myoblast activation or muscle protein synthesis. The muscle activator according to the present invention is highly safe because it is derived from a component of an edible plant.
植物抽出物
ベルガモット(Citrus bergamia)は、ミカン科ミカン属に属する植物である。本発明にかかるベルガモット抽出物には、ベルガモットの各部位、例えば、葉、茎、根、果実、果皮、種子、種皮、地上部、地下部又は全草の抽出物を用いることができる。好ましくは、果皮の抽出物である。
オオバコエンドロ(Eryngium foetidum)は、セリ科ヒゴタイサイ属に属する植物である。本発明にかかるオオバコエンドロ抽出物には、オオバコエンドロの各部位、例えば、葉、茎、根、果実、果皮、種子、種皮、地上部、地下部又は全草の抽出物を用いることができる。好ましくは、全草の抽出物である。
ニイタカヨモギ(Artemisia campestris)は、キク科ヨモギ属に属する植物である。本発明にかかるニイタカヨモギ抽出物には、ニイタカヨモギの各部位、例えば、葉、茎、枝、根、果実、果皮、種子、種皮、地上部、地下部又は全草の抽出物を用いることができる。好ましくは、小枝と葉の抽出物である。
ワダソウ(Pseudostellaria heterophylla)は、ナデシコ科ワチガイソウ属に属する植物である。本発明にかかるワダソウ抽出物には、ワダソウの各部位、例えば、葉、茎、根、果実、果皮、種子、種皮、地上部、地下部又は全草の抽出物を用いることができる。好ましくは、根の抽出物である。
トゲバンレイシ(Annona muricata)は、バンレイシ科バンレイシ属に属する植物である。本発明にかかるトゲバンレイシ抽出物には、トゲバンレイシの各部位、例えば、葉、茎、根、果実、果皮、種子、種皮、地上部、地下部又は全草の抽出物を用いることができる。好ましくは、果実の抽出物である。
キリンケツ(Daemonorops draco)は、ヤシ科ヒメトウ属に属する植物である。本発明にかかるキリンケツ抽出物には、キリンケツの各部位、例えば、葉、茎、根、果実、果皮、種子、種皮、地上部、地下部又は全草の抽出物を用いることができる。好ましくは、果実の抽出物である。
ワニナシ(Persea americana)は、クスノキ科ワニナシ属に属する植物の抽出物である。本発明にかかるワニナシ抽出物には、ワニナシの各部位、例えば、葉、茎、根、果実、果皮、種子、種皮、地上部、地下部又は全草の抽出物を用いることができる。好ましくは、果実の抽出物である。
ベニバナ(Carthamus tinctorius)は、キク科ベニバナ属に属する植物の抽出物である。本発明にかかるベニバナ抽出物には、ベニバナの各部位、例えば、花、葉、茎、根、果実、果皮、種子、種皮、地上部、地下部又は全草の抽出物を用いることができる。好ましくは、花の抽出物である。
トウモロコシ(Zea mays)は、イネ科トウモロコシ属に属する植物である。本発明にかかるトウモロコシ抽出物には、トウモロコシの各部位、例えば、めしべ(ひげ)、茎、根、果実、果皮、種子、種皮、地上部、地下部又は全草の抽出物を用いることができる。好ましくは、めしべの抽出物である。
ツルナ(Tetragonia tetragonioides)は、ハマミズナ科ツルナ属に属する植物である。本発明にかかるツルナ抽出物には、ツルナの各部位、例えば、葉、茎、根、果実、果皮、種子、種皮、地上部、地下部又は全草の抽出物を用いることができる。好ましくは、全草の抽出物である。
ソバ(Fagopyrum esculentum)は、タデ科ソバ属に属する植物である。本発明にかかるソバ抽出物には、ソバの各部位、例えば、葉、茎、根、果実、果皮、種子、種皮、地上部、地下部又は全草の抽出物を用いることができる。好ましくは、種子の抽出物である。
ウド(Aralia cordata)は、ウコギ科タラノキ属に属する植物である。本発明にかかるウド抽出物には、ウドの各部位、例えば、葉、花、茎、根、果実、果皮、種子、種皮、地上部、地下部又は全草の抽出物を用いることができる。好ましくは、茎及び葉の抽出物である。
キカラスウリ(Trichosanthes kirilowii var. japonica)は、ウリ科カラスウリ属に属する植物である。本発明にかかるキカラスウリ抽出物には、キカラスウリの各部位、例えば、葉、茎、根、果実、果皮、種子、種皮、地上部、地下部又は全草の抽出物を用いることができる。好ましくは、種子の抽出物である。
Plant extract bergamot is a plant belonging to the genus Citrus mandarin. As the bergamot extract according to the present invention, extracts of various parts of bergamot, for example, leaves, stems, roots, fruits, fruit skins, seeds, seed coats, above-ground parts, underground parts, or whole grasses can be used. Preferably, it is a fruit skin extract.
Psyllium (Eryngium foetidum) is a plant belonging to the genus Syphaceae. For the psyllium extract according to the present invention, extracts of psyllium, such as leaves, stems, roots, fruits, pericarps, seeds, seed coats, above-ground parts, underground parts, or whole plants can be used. Preferably, it is a whole plant extract.
Nitta hawk mugwort (Artemisia campestris) is a plant belonging to the genus Artemisia mugwort. For the Niitake sagebrush extract according to the present invention, it is possible to use each part of Niitake sagebrush, for example, an extract of leaves, stems, branches, roots, fruits, peels, seeds, seed coats, aerial parts, underground parts or whole grasses. it can. Preferably, it is a twig and leaf extract.
Wadder (Pseudostellaria heterophylla) is a plant belonging to the genus Lacidae. As the Wadder extract according to the present invention, an extract of each part of Wadder, for example, leaf, stem, root, fruit, fruit skin, seed, seed coat, above-ground part, underground part or whole plant can be used. Preferably, it is a root extract.
Togebanreishi (Annona muricata) is a plant belonging to the genus Vanleci. For the spider banci extract according to the present invention, extracts of the spider basilis, for example, leaves, stems, roots, fruits, pericarps, seeds, seed coats, above-ground parts, underground parts, or whole grasses can be used. Preferably, it is a fruit extract.
The giraffe (Daemonorops draco) is a plant belonging to the genus Hymenoptera. For the giraffe extract according to the present invention, it is possible to use extracts of various parts of the giraffe, for example, leaves, stems, roots, fruits, pericarps, seeds, seed coats, above-ground parts, underground parts or whole grasses. Preferably, it is a fruit extract.
Crocodile (Persea americana) is an extract of a plant belonging to the genus Allenaceae. For the crocodile extract according to the present invention, extracts of each part of the crocodile, for example, leaves, stems, roots, fruits, pericarps, seeds, seed coats, above-ground parts, underground parts or whole grasses can be used. Preferably, it is a fruit extract.
Safflower (Carthamus tinctorius) is an extract of a plant belonging to the genus Asteraceae. For the safflower extract according to the present invention, extracts of safflower parts, such as flowers, leaves, stems, roots, fruits, pericarps, seeds, seed coats, above-ground parts, underground parts, or whole grasses, can be used. Preferably, it is a flower extract.
Maize (Zea mays) is a plant belonging to the genus Cornaceae. For the corn extract according to the present invention, extracts of various parts of corn, for example, pistil (beard), stem, root, fruit, peel, seed, seed coat, above-ground part, underground part or whole plant can be used. . Preferably, it is a pistil extract.
The tuna (Tetragonia tetragonioides) is a plant belonging to the genus Tsuruna. For the tsuruna extract according to the present invention, extracts of tsuruna parts such as leaves, stems, roots, fruits, fruit skins, seeds, seed coats, above-ground parts, underground parts or whole plants can be used. Preferably, it is a whole plant extract.
Buckwheat (Fagopyrum esculentum) is a plant belonging to the genus Buckwheat. For the buckwheat extract according to the present invention, the extract of each part of buckwheat, for example, leaf, stem, root, fruit, fruit skin, seed, seed coat, above-ground part, underground part or whole plant can be used. Preferably, it is a seed extract.
Udo (Aralia cordata) is a plant belonging to the genus Arachnaceae. As the udo extract according to the present invention, extracts of various parts of udo, for example, leaves, flowers, stems, roots, fruits, fruit skins, seeds, seed coats, above-ground parts, underground parts, or whole grasses can be used. Preferably, it is a stem and leaf extract.
Kikarasuri (Trichosanthes kirilowii var. Japonica) is a plant that belongs to the genus Lilyaceae. As the extract of citrus cucumber according to the present invention, an extract of each part of citrus cucumber, for example, leaf, stem, root, fruit, pericarp, seed, seed coat, above-ground part, underground part or whole plant can be used. Preferably, it is a seed extract.
本発明にかかる植物抽出物の製造方法は、特に限定されず、通常用いられる方法により製造することができる。例えば、植物の各部位をそのまま又は適当な大きさに切断し、必要に応じて乾燥し、搾汁又は溶媒で抽出することにより製造することができる。
抽出溶媒としては、特に限定されないが、例えば、水、メタノール、エタノール、プロパノール、イソプロパノール、n−ブタノール等の低級アルコール、1,3−ブチレングリコール、プロピレングリコール、グリセリン等の多価アルコール、エチルエーテル、プロピルエーテル等のエーテル類、酢酸エチル等のエステル類、アセトン、エチルメチルケトン等のケトン類を挙げることができる。また、これらの溶媒を一種又は二種以上混合して用いることもできる。抽出溶媒の中で、特に、水、メタノール、エタノール、含水エタノールが好ましい。
The manufacturing method of the plant extract concerning this invention is not specifically limited, It can manufacture by the method used normally. For example, it can be produced by cutting each part of a plant as it is or by cutting it into an appropriate size, drying it if necessary, and extracting it with juice or a solvent.
Although it does not specifically limit as an extraction solvent, For example, lower alcohols, such as water, methanol, ethanol, propanol, isopropanol, n-butanol, polyhydric alcohols, such as 1, 3- butylene glycol, propylene glycol, and glycerol, ethyl ether, Mention may be made of ethers such as propyl ether, esters such as ethyl acetate, and ketones such as acetone and ethyl methyl ketone. These solvents can be used alone or in combination. Among the extraction solvents, water, methanol, ethanol, and water-containing ethanol are particularly preferable.
抽出に使用する溶媒の量は、抽出条件等により異なるが、重量比で1:2〜1:30(植物:溶媒)の範囲内が適当であり、1:3〜1:20の範囲内が好ましく、1:5〜1:10の範囲内がより好ましい。抽出時間は、1時間〜15日の範囲内が適当である。抽出温度は、5〜100℃の範囲内が適当であり、60〜100℃の範囲内が好ましい。
抽出方法については特に制限されず、バッチ抽出、カラムを用いた連続抽出等、任意の方法を適用することができる。
The amount of the solvent used for the extraction varies depending on the extraction conditions and the like, but the weight ratio is suitably in the range of 1: 2 to 1:30 (plant: solvent), and in the range of 1: 3 to 1:20. The range of 1: 5 to 1:10 is more preferable. The extraction time is suitably in the range of 1 hour to 15 days. The extraction temperature is suitably in the range of 5 to 100 ° C, and preferably in the range of 60 to 100 ° C.
The extraction method is not particularly limited, and any method such as batch extraction or continuous extraction using a column can be applied.
得られた抽出物は、そのままの状態で用いることもできるし、必要に応じ、その活性に影響のない範囲内で更に精製処理を加えてもよい。かかる精製処理は、通常の方法によって行えばよく、例えば、抽出物を常法によりろ過することにより行うことができる。また、得られたろ液を減圧濃縮、凍結乾燥又はスプレードライヤーに供することもできる。 The obtained extract can be used as it is, or if necessary, further purification treatment may be added within a range that does not affect its activity. Such purification treatment may be performed by a usual method, for example, by filtering the extract by a conventional method. Moreover, the obtained filtrate can also be used for vacuum concentration, freeze-drying, or a spray dryer.
筋肉活性化剤
本発明にかかる筋肉活性化剤(以下、「本発明筋肉活性化剤」という。)は、筋芽細胞賦活作用又はp70S6K活性化作用、rpS6活性化作用を有し、筋肉の増量、増強や筋萎縮抑制作用、運動による筋肉損傷からの回復促進作用、運動不足や老化に伴うサルコペニア及びロコモティブシンドロームの予防及び治療等に有用である。また、本発明筋肉活性化剤は、皮膚領域近傍に存在する筋芽細胞を活性化し、さらには筋タンパク質の合成を促進することでその周辺の皮膚の弾力低下、たるみやしわ等を改善する皮膚改善効果や、疲労の軽減作用、筋肉疲労緩和による肩こり及び眼精疲労軽減作用も期待される。本発明に係る筋肉活性化剤は、特に、筋肉増強剤、筋肉減衰予防剤、皮膚改善剤としての効果が期待される。
本発明筋肉活性化剤は、上記のようにして得られた本発明抽出物をそのまま又は担体として使用することのできる素材と混合し、次いで、常法により粉末状、塊状、液状などの各種形態に加工することにより製造することができる。
Muscle Activator The muscle activator according to the present invention (hereinafter referred to as “the muscle activator of the present invention”) has a myoblast activation action, a p70S6K activation action, and an rpS6 activation action, and increases muscle mass. It is useful for enhancing and preventing muscle atrophy, promoting recovery from muscle damage caused by exercise, preventing and treating sarcopenia and locomotive syndrome associated with lack of exercise and aging. Further, the muscle activating agent of the present invention activates myoblasts existing in the vicinity of the skin region, and further promotes the synthesis of muscle protein, thereby improving the skin elasticity, sagging and wrinkles around the skin. It is also expected to have an improving effect, a fatigue-reducing action, a stiff shoulder by reducing muscle fatigue, and an eye strain-reducing action. The muscle activator according to the present invention is expected to be particularly effective as a muscle strengthening agent, a muscle attenuation preventing agent, and a skin improving agent.
The muscle activating agent of the present invention is prepared by mixing the extract of the present invention obtained as described above with a raw material that can be used as it is or as a carrier, and then in various forms such as powder, lump, and liquid by conventional methods. It can manufacture by processing.
本発明筋肉活化剤における本発明抽出物の含有量は、筋芽細胞賦活作用又はp70S6K活性化作用、rpS6活性化作用が確認できる範囲であれば特に限定されないが、例えば、0.01重量%以上であり、好ましくは、0.05重量%以上、20重量%以下、特に好ましくは、0.1重量%以上、5重量%以下である。 The content of the extract of the present invention in the muscle activating agent of the present invention is not particularly limited as long as the myoblast activation action, the p70S6K activation action, and the rpS6 activation action can be confirmed. For example, 0.01% by weight or more Preferably, they are 0.05 weight% or more and 20 weight% or less, Most preferably, they are 0.1 weight% or more and 5 weight% or less.
本発明筋肉活性化剤には、医薬上又は食品上許容される添加物を任意に配合することができる。かかる添加剤としては、例えば、結合剤、崩壊剤、滑沢剤、分散剤、懸濁剤、乳化剤、緩衝剤、酸化防止剤、賦形剤、界面活性剤、紫外線防止剤、金属イオン封鎖剤、増粘剤、防腐剤、抗菌剤、保湿剤、色素を挙げることができ、これらを1種又は2種以上使用することができる。本発明筋肉活性化剤には、さらに、筋肉増強作用又は皮膚改善作用が知られている成分を添加することができる。例として、筋肉増強作用成分としては、クレアチン、BCAAやロイシンなどのアミノ酸、HMB(3−ヒドロキシイソ吉草酸)、皮膚改善作用としては、コラーゲン、ヒアルロン酸、エラスチン、プロテオグリカン、セラミド、ビタミンCが挙げられる。
本発明筋肉活性化剤は、常法により、錠剤、散剤、顆粒剤、カプセル剤、液剤、シロップ剤、ドロップ錠、トニック、ローション、軟膏、クリーム等の剤型に適宜調製することができる。
本発明筋肉活性化剤の摂取量は、症状、剤型、投与対象者の年齢、体重等により異なるが、通常、成人1日当り、本発明抽出物の重量として、0.01〜100gの範囲内とするのが適当であり、0.03g〜60gの範囲内とするのが好ましく、0.1g〜10gの範囲内とするのが好ましいが、必ずしもこの範囲に限られるものではない。かかる1日当りの摂取量は、1回で摂取してもよく、また、2〜4回に分割して摂取してもよい。
The muscular activator of the present invention can optionally contain a pharmaceutically or food acceptable additive. Examples of such additives include binders, disintegrants, lubricants, dispersants, suspending agents, emulsifiers, buffers, antioxidants, excipients, surfactants, UV inhibitors, and sequestering agents. , Thickeners, preservatives, antibacterial agents, moisturizers, and pigments, and one or more of these can be used. The muscle activator of the present invention may further contain components known to have muscle strengthening action or skin improvement action. Examples include muscle strengthening components such as creatine, amino acids such as BCAA and leucine, HMB (3-hydroxyisovaleric acid), and skin improving effects such as collagen, hyaluronic acid, elastin, proteoglycan, ceramide, and vitamin C. It is done.
The muscle activator of the present invention can be suitably prepared into dosage forms such as tablets, powders, granules, capsules, solutions, syrups, drop tablets, tonics, lotions, ointments, creams and the like by conventional methods.
The intake amount of the muscle activator of the present invention varies depending on the symptoms, dosage form, age of the administration subject, body weight, etc., but usually within the range of 0.01 to 100 g as the weight of the extract of the present invention per adult day. It is suitable to be within the range of 0.03 g to 60 g, and preferably within the range of 0.1 g to 10 g, but is not necessarily limited to this range. Such daily intake may be taken once, or may be taken divided into 2 to 4 times.
以下に、実施例、試験例を掲げて本発明をさらに詳述する。但し、本発明が下記実施例に限定されないことは言うまでもない。 Hereinafter, the present invention will be described in more detail with reference to examples and test examples. However, it goes without saying that the present invention is not limited to the following examples.
製造例1 各植物のメタノール抽出物の製造
表1に記載の各植物の乾燥物15gを粉砕後、メタノール75gに7日間浸漬させ、ろ過後にロータリーエバポレーターにて濃縮・乾固を行った。その後、Dimethyl sulfoxide(DMSO)で終濃度50mg/mLに調製し、各植物のメタノール抽出物を得た。
製造例2 各植物のエタノール抽出物の製造
表1に記載の各植物の乾燥物15gを粉砕後、エタノール75gに7日間浸漬させ、ろ過後にロータリーエバポレーターにて濃縮・乾固を行った。その後、Dimethyl sulfoxide(DMSO)で終濃度50mg/mLに調製し、各植物のエタノール抽出物を得た。
製造例3 各植物の熱水抽出物の製造
表1に記載の各植物の乾燥物10gを粉砕後、100℃の熱水150mLに添加し、30分加熱し、ろ過後にロータリーエバポレーターにて濃縮・乾固を行った。その後、Dimethyl sulfoxide(DMSO)で終濃度50mg/mLに調製し、各植物の熱水抽出物を得た。
Production Example 1 Production of Methanol Extract of Each Plant 15 g of the dried product of each plant shown in Table 1 was pulverized and then immersed in 75 g of methanol for 7 days, and after filtration, concentrated and dried by a rotary evaporator. Thereafter, the final concentration was adjusted to 50 mg / mL with dimethylsulfoxide (DMSO) to obtain a methanol extract of each plant.
Production Example 2 Production of ethanol extract of each plant 15 g of the dried product of each plant shown in Table 1 was pulverized and then immersed in 75 g of ethanol for 7 days, and after filtration, concentrated and dried by a rotary evaporator. Thereafter, the final concentration was adjusted to 50 mg / mL with dimethylsulfoxide (DMSO) to obtain an ethanol extract of each plant.
Production Example 3 Production of hot water extract of each plant 10 g of the dried product of each plant listed in Table 1 was pulverized, then added to 150 mL of hot water at 100 ° C., heated for 30 minutes, filtered and concentrated on a rotary evaporator. Drying was performed. Then, it adjusted to final concentration 50mg / mL with dimethylsulfoxide (DMSO), and the hot water extract of each plant was obtained.
試験例1 本発明にかかる植物抽出物の筋芽細胞賦活活性測定
細胞はマウス筋芽細胞株 (C2C12細胞、DSファーマバイオメディカル社製 )を用いた。C2C12細胞は、10% FBS(Fetal Bovine Serum、Gibco社製)を含有するDMEM(Dulbecco’s Modified Eagle Medium、Gibco社製)(以下、「10% FBS/DMEM培地」という。)に分散し、接着系細胞用培養フラスコ(底面積:25cm2、日本バイオサイエンス社製)に5.0 × 104個播種し、CO2インキュベーター(三洋電機バイオメディカ社製;CO2濃度5%, 湿度90%)を用いて37℃で3日間培養した。80%コンフルエントになったことを顕微鏡で確認し、実験に使用した。80%コンフルエントになったC2C12細胞をTrypsin−EDTA(Gibco社製)を用いてフラスコから剥離させ回収した。この細胞を10%FBS/DMEM培地に懸濁し、3.0×104 cells/mLの細胞密度で100 μLずつ96 well plate(Corning社製)に播種した。37℃、5%CO2条件下で24時間培養後、培地をアスピレーターで除去し、2% FBS/DMEM培地で調製したサンプル含有培地200μLに置換した。サンプルは、製造例1で調製した表1に記載のNo.1〜13の各メタノール抽出物を終濃度の1000倍濃度でDMSOを用いて調整し、その後2%FBS/DMEM培地で1000倍希釈し、サンプル濃度が0.5、1、5、10、20ppmの培地をそれぞれ調製し、使用した。また、コントロールにはDMSOを、陽性対照には、筋芽細胞賦活作用が知られているIGF−1(Insulin−likeGrowth Factor−1、LifeTechnologies社製)を同様のサンプル調製法により0.05ppmで調整し、実験に用いた。サンプル添加の48時間後に100μLの無血清DMEM培地に交換し、そこにWST−1試薬(ロシュ・ダイアグノスティックス社製)を10μL添加した。30分間培養を行った後、450nmの吸光値をマイクロプレートリーダー(CORONA ELECTRIC 社製、MTP−300)で測定することにより細胞賦活活性を求めた。結果を図1〜8に示す。
図1〜8から明らかなように、薬物を添加しないコントロール群の細胞賦活活性を100%とした場合と比較して、各植物抽出物を添加した場合に細胞賦活活性の増加が認められた。
Test Example 1 Measurement of Myoblast Activation Activity of Plant Extract According to the Present Invention A mouse myoblast cell line (C2C12 cell, manufactured by DS Pharma Biomedical Co., Ltd.) was used as the cell. C2C12 cells are dispersed in DMEM (Dulbecco's Modified Eagle Medium, manufactured by Gibco) (hereinafter referred to as “10% FBS / DMEM medium”) containing 10% FBS (Fetal Bovine Serum, manufactured by Gibco). Inoculation cells (bottom area: 25 cm 2 , manufactured by Nippon Biosciences) 5.0 × 10 4 seeded cells, CO 2 incubator (manufactured by Sanyo Biomedica Co., Ltd .; CO 2 concentration 5%,
As is apparent from FIGS. 1 to 8, an increase in cell activation activity was observed when each plant extract was added, compared to the case where the cell activation activity of the control group to which no drug was added was 100%.
試験例2 本発明にかかる植物抽出物のp70S6Kのリン酸化測定
細胞はマウス筋芽細胞株(C2C12細胞、DSファーマバイオメディカル社製)を用いた。C2C12細胞は、10%FBS(Fetal Bovine Serum、Gibco社製)を含有するDMEM (Dulbecco’sModified Eagle Medium、Gibco社製)(以下、「10% FBS/DMEM培地」という。)に分散し、接着系細胞用培養フラスコ(底面積:25cm2、日本バイオサイエンス社製)に5.0 × 104個播種し、CO2インキュベーター(三洋電機バイオメディカ社製;CO2濃度5%, 湿度90%)を用いて37℃で3日間培養した。80%コンフルエントになったことを顕微鏡で確認し、実験に使用した。80%コンフルエントになったC2C12細胞をTrypsin−EDTA(Gibco社製)を用いてフラスコから剥離させ回収した。この細胞を10%FBS/DMEM培地に懸濁し、1.7×105個/wellで12 wellplateに播種した。播種24時間後に2%HS(Horse Serum、Gibco製)1mLに置換し、筋管細胞へと分化させた。分化開始から2日に1回2%HSに置換し、分化3日目の筋管細胞を実験に用いた。分化3日目の筋管細胞を無血清培地に置換した後、9時間後に被験物質として、製造例1で調製した表1に記載のNo.1〜4、13の各メタノール抽出物を1、10、50ppm含んだ無血清培地1mLに置換し、検体処置を行った。処置開始から2時間後にPBS(Phosphatebuffered saline)で洗浄した後、RIPA buffer(富士フィルム和光純薬社製)20μLで細胞を回収した。得られたタンパク溶液を用いて、SDS−PAGEサンプルバッファーに添加し、100℃、3分で還元処理を行い、続くウエスタンブロットにてタンパク量50μgを10%アクリルアミドゲルで分離した後、p70S6Kのリン酸化の定量を行った。なお、抗体はAnti−p70S6K及びAnti−phospho−p70S6K(Cell Signaling Technology社製)を用いた。結果を図9〜13に示す。
図9〜13から明らかなように、本発明にかかるベルガモット抽出物、オオバコエンドロ抽出物、ニイタカヨモギ抽出物、ワダソウ抽出物およびキカラスウリ抽出物は、p70S6Kのリン酸化を促進した。
Test Example 2 Measurement of phosphorylation of p70S6K in the plant extract according to the present invention The mouse myoblast cell line (C2C12 cell, manufactured by DS Pharma Biomedical Co., Ltd.) was used. C2C12 cells are dispersed in DMEM (Dulbecco's Modified Eagle Medium, Gibco) containing 10% FBS (Fetal Bovine Serum, Gibco) (hereinafter referred to as “10% FBS / DMEM medium”). Cell culture flasks (bottom area: 25 cm 2 , manufactured by Nippon Biosciences Co., Ltd.) were seeded with 5.0 × 10 4 cells, and CO 2 incubator (manufactured by Sanyo Biomedica Co., Ltd .; CO 2 concentration 5%,
As is clear from FIGS. 9 to 13, the bergamot extract, psyllium endosperm extract, Niitake sagebrush extract, Wadder extract and red pepper extract according to the present invention promoted phosphorylation of p70S6K.
試験例3 本発明にかかる植物抽出物のrpS6のリン酸化測定
抗体として、Anti−rpS6K及びAnti−phospho−rpS6K(Cell Signaling Technology社製)を用いること以外は、試験例2と同様の方法を用いてrpS6Kのリン酸化を定量することができる。
Test Example 3 rpS6 Phosphorylation Measurement of Plant Extract According to the Present Invention The same method as in Test Example 2 was used except that Anti-rpS6K and Anti-phospho-rpS6K (manufactured by Cell Signaling Technology) were used as antibodies. Thus, phosphorylation of rpS6K can be quantified.
本発明にかかる筋肉活性化剤は、優れた筋肉活性化作用を有するため、例えば、運動不足や老化等に伴うサルコペニア、ロコモティブシンドローム、様々な環境(ギプス固定等)による廃用性委縮、又はロコモティブシンドロームの予防及び改善、運動による筋肉増強の強化、皮膚の弾力低下、しわやたるみの抑制に用いることができる。 Since the muscle activating agent according to the present invention has an excellent muscle activating action, for example, sarcopenia associated with lack of exercise, aging, etc., locomotive syndrome, disuse atrophy due to various environments (such as cast fixation), or locomotive It can be used for the prevention and improvement of syndrome, the enhancement of muscle enhancement by exercise, the reduction of skin elasticity, and the suppression of wrinkles and sagging.
Claims (3)
The muscle activator according to claim 1, wherein the muscle activator is a skin improving agent.
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JP2010195696A (en) * | 2009-02-24 | 2010-09-09 | Ichimaru Pharcos Co Ltd | PREPARATION FOR ACTIVATING Akt-mTOR-p70S6K OF MUSCLE STRENGTHENING SYSTEM AND METHOD THEREOF |
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