JP2020100591A - Agent for improving muscle quality - Google Patents
Agent for improving muscle quality Download PDFInfo
- Publication number
- JP2020100591A JP2020100591A JP2018240019A JP2018240019A JP2020100591A JP 2020100591 A JP2020100591 A JP 2020100591A JP 2018240019 A JP2018240019 A JP 2018240019A JP 2018240019 A JP2018240019 A JP 2018240019A JP 2020100591 A JP2020100591 A JP 2020100591A
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- JP
- Japan
- Prior art keywords
- muscle
- agent
- present
- fucoxanthin
- exercise
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
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Abstract
Description
本発明は、運動の効果を高め、筋質を向上させ得る筋質向上剤に関する。 The present invention relates to a muscle quality improving agent that can enhance the effect of exercise and improve muscle quality.
近年、高齢化が進むのに伴い、メタボリックシンドローム(代謝症候群)、ロコモティブシンドローム(運動器症候群)、フレイル(健康な状態と要介護状態の中間の状態)といった身体機能の低下を伴う健康障害が社会問題となっている。
これに対して従来は、骨格筋量を増加させることを主眼に様々な取り組みが進められてきたが、骨格筋量の増加だけでは、筋力の回復には不十分であることが明らかになってきた。
また、これまでは、加齢による変化として捉えられていた筋肉の質、すなわち筋質の低下が、バランスの偏った食事や運動不足によっても惹起され、将来的には、ロコモティブシンドロームやフレイル等の深刻な状態に至ることがわかってきた。
かかる筋質の低下を改善し、または予防するには、運動が最も有効な手段であることが知られている。しかし、運動を行いたくても、加齢による身体機能の低下、疾患、負傷等による運動の制限等、種々の原因により、十分な運動を実施できないことも多い。
そこで、筋質の低下を予防し、また筋質を向上させることができ、無理のない範囲で実施できる運動であっても、その効果を有効に増強させ得る筋質向上剤の開発が望まれている。
With the aging of the population in recent years, social disorders such as metabolic syndrome (locomotive syndrome), locomotive syndrome (motor organ syndrome), and frail (a state intermediate between a healthy state and a state requiring long-term care) are associated with social disorders. It's a problem.
On the other hand, in the past, various efforts have been made with a focus on increasing skeletal muscle mass, but it has become clear that increasing skeletal muscle mass alone is not sufficient for recovery of muscle strength. It was
In addition, until now, the deterioration of muscle quality, that is, the deterioration of muscle quality, which has been regarded as a change due to aging, is also caused by imbalanced diet and lack of exercise, and in the future, such as locomotive syndrome and flail. It has become clear that it will lead to a serious situation.
It is known that exercise is the most effective means for improving or preventing such muscle deterioration. However, even if it is desired to perform exercise, it is often impossible to perform sufficient exercise due to various causes such as deterioration of physical function due to aging, limitation of exercise due to diseases and injuries.
Therefore, it is desirable to develop a muscle quality improving agent that can prevent muscle deterioration and improve muscle quality, and that can effectively enhance the effect of exercise even if it can be exercised within a reasonable range. ing.
ところで、昨今の健康志向の高まりにより、食品素材をはじめ可食性の成分に種々の生理活性を求める研究が進められている。
たとえば、褐藻類に含まれるカロテノイドであるフコキサンチンについて、脂肪の燃焼促進作用や抗腫瘍作用が報告され、糖尿病治療剤への応用も検討されている(特許文献1)。
しかしながら、フコキサンチンが筋質をはじめ筋肉の機能に及ぼす影響については、これまで報告されていない。
By the way, with the recent increase in health consciousness, researches for various physiological activities of edible ingredients including food materials are being promoted.
For example, fucoxanthin, a carotenoid contained in brown algae, has been reported to have a fat burning promoting action and an antitumor action, and its application to a therapeutic agent for diabetes has also been investigated (Patent Document 1).
However, the effect of fucoxanthin on muscle function including muscle quality has not been reported so far.
そこで、本発明は、運動が制限される場合であっても、筋質の低下を予防し、また筋質を向上させることができ、さらに、無理のない程度の運動であっても、その効果を有効に増強させ得る筋質向上剤を提供することを目的とした。 Therefore, the present invention can prevent deterioration of muscle quality and improve muscle quality even when exercise is limited. Furthermore, even if exercise is reasonably performed, its effect is obtained. The purpose of the present invention is to provide a muscle-improving agent that can effectively enhance the muscle.
上記課題を解決すべく、本発明者らは鋭意検討した結果、フコキサンチンおよびその誘導体からなる群より選択される1種または2種以上が、筋質の低下を予防し、または筋質を向上させる効果を有することを見出し、本発明を完成するに至った。 In order to solve the above-mentioned problems, as a result of intensive investigations by the present inventors, one or two or more kinds selected from the group consisting of fucoxanthin and its derivatives prevent deterioration of muscle quality or improve muscle quality. The inventors have found that it has the effect of making the present invention complete the present invention.
すなわち、本発明は以下に関する。
[1]フコキサンチンおよびその誘導体からなる群より選択される1種または2種以上を含有する、筋質向上剤。
[2]フコキサンチンの誘導体がフコキサンチノールである、[1]に記載の筋質向上剤。
[3]フコキサンチンおよびその誘導体からなる群より選択される1種または2種以上を1nM〜1mM含有する、[1]または[2]に記載の筋質向上剤。
[4][1]〜[3]のいずれかに記載の筋質向上剤を含有する、筋質向上用の医薬。
[5][1]〜[3]のいずれかに記載の筋質向上剤を含有する、筋質向上用の食品。
That is, the present invention relates to the following.
[1] A muscle improving agent containing one or more selected from the group consisting of fucoxanthin and its derivatives.
[2] The muscle improving agent according to [1], wherein the fucoxanthin derivative is fucoxanthinol.
[3] The muscle-improving agent according to [1] or [2], containing 1 nM to 1 mM of one or more selected from the group consisting of fucoxanthin and its derivatives.
[4] A medicine for improving muscle quality, containing the muscle improving agent according to any one of [1] to [3].
[5] A food for improving muscle quality, containing the muscle improving agent according to any one of [1] to [3].
本発明の筋質向上剤は、運動が制限される場合であっても、加齢等、種々の原因による筋質の低下を予防し、また、筋質を向上させることができる。さらに、無理のない程度の運動であっても、その効果を有効に増強させることができる。
従って、加齢による身体機能の低下や、疾患、負傷等による運動の制限等により、従来筋質を向上させるのに必要とされていた運動を実施することが困難な者においても、筋質の低下を予防し、筋質を向上させるのに有用である。
The muscle mass improving agent of the present invention can prevent muscle mass deterioration due to various causes such as aging and improve muscle mass even when exercise is restricted. Furthermore, the effect can be effectively enhanced even if the exercise is reasonably done.
Therefore, even if it is difficult to perform the exercise that was conventionally required to improve the muscle quality due to deterioration of physical function due to aging, limitation of exercise due to diseases, injuries, etc. It is useful for preventing decline and improving muscle quality.
本発明は、筋質向上剤を提供する。
本発明の筋質向上剤(以下、本明細書において「本発明の剤」とも称する)は、フコキサンチンおよびその誘導体からなる群より選択される1種または2種以上を含有する。
The present invention provides a muscle improving agent.
The muscle-improving agent of the present invention (hereinafter, also referred to as "agent of the present invention" in the present specification) contains one or more selected from the group consisting of fucoxanthin and its derivatives.
ここで、「筋質」とは、上述した通り、筋肉の質、つまり筋肉の状態を意味する。筋肉は、筋線維と、水、脂肪、結合組織等の筋線維をとりまく組織の集合体であり、筋質が良好であると評価される筋肉では、筋線維が密に存在し、筋線維をとりまく組織が少ない状態であるが、筋線維の数が減少し、または筋線維が細くなり、筋線維以外の組織の割合が増加すると、筋質が低下したと評価される。これに加えて、筋ミトコンドリア機能、筋肉のインスリン感受性、筋肉の炎症、脂質代謝、タンパク質合成などの筋肉の有する機能も筋質の要素である。たとえば、筋肉が衰えてくると、脂質代謝機能が低下して脂肪が溜まりやすくなり、筋ミトコンドリア機能や筋タンパク質合成機能が低下するが、かかる機能の低下によっても、筋質が低下したと評価される。
それゆえ、「筋質の低下の予防」とは、筋線維が減少すること、筋線維が細くなること、筋線維の周りの脂肪や結合組織が増加することを防ぎ、また、筋ミトコンドリア機能を維持すること、筋肉のインスリン感受性を維持すること、筋肉の炎症や、筋肉の脂質代謝機能、タンパク質合成機能の低下を防ぐことをいい、「筋質の向上」とは、筋線維を増加させ、または筋線維を太くして密な状態とし、筋線維の周りの脂肪や結合組織を減少させ、筋肉の炎症やインスリン感受性を改善し、筋ミトコンドリア機能や脂質代謝機能、タンパク質合成機能等、筋肉の本来の代謝的機能を向上させて筋肉を良好な状態とすることをいう。
本発明における「筋質向上剤」は、上記した筋質の低下を予防する機能を有するもの、または筋質を向上させる機能を有するもの、あるいはそれらの両方の機能を有するものをいう。
Here, the "muscle quality" means the quality of muscles, that is, the state of muscles, as described above. A muscle is an aggregate of muscle fibers and tissues surrounding the muscle fibers such as water, fat, and connective tissue.In a muscle evaluated to have good muscle quality, the muscle fibers are densely present and When the number of surrounding tissues is small, but the number of muscle fibers decreases or the muscle fibers become thin and the ratio of tissues other than muscle fibers increases, it is evaluated that muscle quality is deteriorated. In addition to this, muscle functions such as muscle mitochondrial function, muscle insulin sensitivity, muscle inflammation, lipid metabolism, and protein synthesis are also muscle elements. For example, when muscle declines, lipid metabolism is reduced and fat is more likely to accumulate, which reduces muscle mitochondrial function and muscle protein synthesis function. It
Therefore, "prevention of muscle deterioration" means to prevent the decrease of muscle fibers, the thinning of muscle fibers, the increase of fat and connective tissue around the muscle fibers, and the function of muscle mitochondria. To maintain, to maintain the insulin sensitivity of muscle, to prevent the inflammation of muscle and the decrease of muscle lipid metabolism function and protein synthesis function, "improvement of muscle quality" means increasing muscle fiber, Or thicken muscle fibers to make them dense, reduce fat and connective tissue around muscle fibers, improve muscle inflammation and insulin sensitivity, and improve muscle mitochondrial function, lipid metabolism function, protein synthesis function, etc. It means improving the original metabolic function to improve the muscle condition.
The “muscle quality improving agent” in the present invention means one having a function of preventing the above-mentioned deterioration of muscle quality, one having a function of improving muscle quality, or one having both of these functions.
本発明の剤に有効成分として含有されるフコキサンチンは、(1S,4S,6R)−1−[(3E,5E,7E,9E,11E,13E,15E)−18−[(1R,2R,4S)−2,6,6−トリメチル−2−ヒドロキシ−4−アセトキシシクロヘキサン−1−イリデン]−3,7,12,16−テトラメチル−2−オキソ−3,5,7,9,11,13,15,17−オクタデカオクタエン−1−イル]−2,2,6−トリメチル−7−オキサビシクロ[4.1.0]ヘプタン−4−オールであり、褐藻(Phaeophyceae)等の不等毛藻(Heterokontophyta)中に存在する非プロビタミンA類のカロテノイドの1種であり、キサントフィルに属し、アレン構造、エポキシドおよびヒドロキシ基を有する。
フコキサンチンの誘導体としては、フコキサンチンより化学的または生理学的に誘導される誘導体であれば特に限定されないが、フコキサンチンの代謝生成物であるフコキサンチノール等が好ましい例として挙げられる。
フコキサンチンは、褐藻等から抽出し、精製する方法等、公知の方法に従って製造して用いることができ、フコキサンチンの誘導体は、フコキサンチンより、化学的または生理学的に誘導する方法により、製造して用いることができる。また、各社より提供されている市販の製品を用いることもできる。
本発明の剤には、フコキサンチンおよびその誘導体からなる群より1種または2種以上を選択して、含有させることができる。
本発明の剤におけるフコキサンチンおよびその誘導体からなる群より選択される1種または2種以上(以下、本明細書にて「フコキサンチン等」ということがある)の含有量は、通常1nM〜1mMであり、好ましくは5nM〜500μMであり、より好ましくは10nM〜300μMである。
The fucoxanthin contained in the agent of the present invention as an active ingredient is (1S,4S,6R)-1-[(3E,5E,7E,9E,11E,13E,15E)-18-[(1R,2R, 4S)-2,6,6-Trimethyl-2-hydroxy-4-acetoxycyclohexane-1-ylidene]-3,7,12,16-tetramethyl-2-oxo-3,5,7,9,11, 13,15,17-octadecaoctane-1-yl]-2,2,6-trimethyl-7-oxabicyclo[4.1.0]heptan-4-ol, which is a non-toxic agent of brown algae (Phaeophyceae) and the like. It is one of the non-provitamin A carotenoids present in Heterokontophyta, belongs to xanthophyll, and has an allene structure, an epoxide and a hydroxy group.
The derivative of fucoxanthin is not particularly limited as long as it is a derivative chemically or physiologically derived from fucoxanthin, but fucoxanthinol, which is a metabolite of fucoxanthin, and the like are preferable examples.
Fucoxanthin can be produced and used according to a known method such as a method of extracting from brown algae and the like, and a method of purifying, and a derivative of fucoxanthin is produced from fucoxanthin by a method of chemically or physiologically inducing it. Can be used. In addition, commercially available products provided by each company can also be used.
The agent of the present invention may contain one or more selected from the group consisting of fucoxanthin and its derivatives.
In the agent of the present invention, the content of one or more kinds selected from the group consisting of fucoxanthin and its derivatives (hereinafter, sometimes referred to as "fucoxanthin and the like" in the present specification) is usually 1 nM to 1 mM. And is preferably 5 nM to 500 μM, more preferably 10 nM to 300 μM.
本発明の剤には、他の栄養成分や抗疲労剤を含有させることもできる。かかる栄養成分等としては、具体的には、グルコース、デキストラン等の糖質製剤、精製大豆油、精製卵黄レシチン等の脂肪乳剤、カゼイン、ホエイタンパク等のタンパク質製剤、カフェイン、ビタミン類、ミネラル類、ポリフェノール類等が挙げられる。 The agent of the present invention can also contain other nutritional ingredients and anti-fatigue agents. Specific examples of such nutrients include glucose, sugar preparations such as dextran, refined soybean oil, fat emulsion such as refined egg yolk lecithin, casein, protein preparations such as whey protein, caffeine, vitamins and minerals. , Polyphenols and the like.
本発明の剤は、錠剤、被覆錠剤、チュアブル錠、丸剤、(マイクロ)カプセル剤、顆粒剤、細粒剤、散剤、エリキシル剤、リモナーゼ剤、シロップ剤、懸濁剤、乳剤、経口ゼリー剤等の経口製剤、溶液状、懸濁液状、乳液状等の注射剤、用時溶解または懸濁して用いる固形状の注射剤、輸液剤、持続性注射剤等の注射用製剤等の剤形とすることができる。 The agent of the present invention is a tablet, coated tablet, chewable tablet, pill, (micro)capsule, granule, fine granule, powder, elixir, limonase agent, syrup, suspension, emulsion, oral jelly. And other dosage forms such as oral preparations, injections such as solution, suspension, and emulsion, solid injections that are dissolved or suspended before use, infusions, sustained injections, and other injection preparations can do.
上記剤形の本発明の剤は、製剤の分野で周知の製剤化手段、たとえば第十七改正日本薬局方製剤総則[3]製剤各条に記載された方法等により、調製することができる。
その際、必要に応じて、薬理学的に許容し得る各種の製剤用添加剤を配合することができる。当該添加剤は、本発明の剤の剤形に応じて適宜選択することができるが、たとえば、賦形剤、結合剤、崩壊剤、滑沢剤、被覆剤、基剤、溶剤、希釈剤、溶解補助剤、可溶化剤、乳化剤、分散剤、懸濁化剤、安定化剤、粘稠剤、無痛化剤、等張化剤、pH調整剤、抗酸化剤、防腐剤、保存剤、矯味剤、風味剤、甘味剤、香料、着色剤等が挙げられる。
The agent of the present invention in the above dosage form can be prepared by a formulation means well known in the field of formulation, for example, the method described in the 17th Revised Japanese Pharmacopoeia General Provisions [3] Preparation Articles.
At that time, if necessary, various pharmacologically acceptable additives for formulation can be blended. The additive can be appropriately selected according to the dosage form of the agent of the present invention, for example, an excipient, a binder, a disintegrant, a lubricant, a coating agent, a base, a solvent, a diluent, Dissolution aids, solubilizers, emulsifiers, dispersants, suspending agents, stabilizers, thickeners, soothing agents, isotonic agents, pH adjusting agents, antioxidants, preservatives, preservatives, taste masking agents. Agents, flavoring agents, sweetening agents, flavoring agents, coloring agents and the like.
具体的には、たとえば賦形剤としては、炭酸マグネシウム、二酸化チタン、糖類(ラクトース等)、糖アルコール(マンニトール等)、カゼイン等が挙げられる。
結合剤としては、ゼラチン、澱粉、セルロースおよびその誘導体等が挙げられる。
崩壊剤としては、クロスポビドン、結晶セルロース等が挙げられる。
滑沢剤としては、タルク、ステアリン酸マグネシウム等が挙げられる。
被覆剤としては、メタクリル酸メチル・メタクリル酸ブチル・メタクリル酸ジメチルアミノエチル共重合体、アクリル酸エチル・メタクリル酸メチル・メタクリル酸塩化トリメチルアンモニウムエチル共重合体等が挙げられる。
基剤としては、動物油、植物油、炭化水素油(流動パラフィン等)、ポリエチレングリコール等が挙げられる。
溶剤としては、精製水、注射用水、一価アルコール(エタノール等)、多価アルコール(グリセリン等)等が挙げられる。
乳化剤または分散剤としては、ソルビタン脂肪酸エステル、グリセリン脂肪酸エステル、ポリオキシエチレンソルビタン脂肪酸エステル、ショ糖脂肪酸エステル等が挙げられる。
安定化剤としては、アジピン酸、β−シクロデキストリン等が挙げられる。
粘稠剤としては、水溶性高分子(ポリアクリル酸ナトリウム、カルボキシビニルポリマー等)、多糖類(アルギン酸ナトリウム、キサンタンガム、トラガント等)等が挙げられる。
無痛化剤としては、アミノ安息香酸エチル、クロロブタノール、プロピレングリコール、ベンジルアルコール等が挙げられる。
等張化剤としては、塩化カリウム、塩化ナトリウム、ソルビトール、生理食塩液等が挙げられる。
pH調整剤としては、塩酸、硫酸、酢酸、クエン酸、乳酸、水酸化ナトリウム、水酸化カリウム等が挙げられる。
抗酸化剤としては、ジブチルヒドロキシトルエン(BHT)、ブチルヒドロキシアニソール(BHA)、α−トコフェロール、エリソルビン酸等が挙げられる。
防腐剤または保存剤としては、パラベン(メチルパラベン等)、ベンジルアルコール、デヒドロ酢酸ナトリウム、ソルビン酸等が挙げられる。
矯味剤または風味剤としては、アスコルビン酸、エリスリトール、L−グルタミン酸ナトリウム等が挙げられ、甘味剤としては、アスパルテーム、カンゾウエキス、サッカリン等が挙げられる。
香料としては、l−メントール、d−カンファー、シネオール等が挙げられる。
着色剤としては、タール色素(赤色2号、青色1号、黄色4号等)、無機顔料(ベンガラ、黄酸化鉄、黒酸化鉄等)、天然色素(アナトー色素、ウコン色素、β−カロテン等)等が挙げられる。
Specifically, examples of the excipient include magnesium carbonate, titanium dioxide, sugars (lactose and the like), sugar alcohols (mannitol and the like), casein and the like.
Examples of the binder include gelatin, starch, cellulose and its derivatives.
Examples of the disintegrant include crospovidone, crystalline cellulose and the like.
Examples of lubricants include talc and magnesium stearate.
Examples of the coating agent include a methyl methacrylate/butyl methacrylate/dimethylaminoethyl methacrylate copolymer and an ethyl acrylate/methyl methacrylate/trimethylammonium ethyl methacrylate methacrylate copolymer.
Examples of the base include animal oil, vegetable oil, hydrocarbon oil (liquid paraffin, etc.), polyethylene glycol and the like.
Examples of the solvent include purified water, water for injection, monohydric alcohol (ethanol and the like), polyhydric alcohol (glycerin and the like) and the like.
Examples of the emulsifier or dispersant include sorbitan fatty acid ester, glycerin fatty acid ester, polyoxyethylene sorbitan fatty acid ester, and sucrose fatty acid ester.
Examples of the stabilizer include adipic acid and β-cyclodextrin.
Examples of the thickening agent include water-soluble polymers (sodium polyacrylate, carboxyvinyl polymer, etc.), polysaccharides (sodium alginate, xanthan gum, tragacanth, etc.) and the like.
Examples of soothing agents include ethyl aminobenzoate, chlorobutanol, propylene glycol, benzyl alcohol and the like.
Examples of the isotonicity agent include potassium chloride, sodium chloride, sorbitol, physiological saline and the like.
Examples of pH adjusters include hydrochloric acid, sulfuric acid, acetic acid, citric acid, lactic acid, sodium hydroxide, potassium hydroxide and the like.
Examples of the antioxidant include dibutylhydroxytoluene (BHT), butylhydroxyanisole (BHA), α-tocopherol, and erythorbic acid.
Examples of the preservative or preservative include paraben (methylparaben and the like), benzyl alcohol, sodium dehydroacetate, sorbic acid and the like.
Examples of the corrigent or flavoring agent include ascorbic acid, erythritol, sodium L-glutamate, and the like, and examples of the sweetener include aspartame, licorice extract, saccharin, and the like.
Examples of the fragrance include l-menthol, d-camphor and cineol.
Examples of colorants include tar dyes (red No. 2, blue No. 1, yellow No. 4, etc.), inorganic pigments (red iron oxide, yellow iron oxide, black iron oxide, etc.), natural dyes (anato dye, turmeric dye, β-carotene, etc.). ) And the like.
本発明の剤の摂取量または投与量は、本発明の剤が適用される対象(以下、本明細書において「適用対象」ともいう)における筋質の状態または筋質低下の程度、性別、年齢、体重、剤形、投与方法等により適宜決定されるが、適用対象がヒト成人である場合、フコキサンチンに換算した量として、1日あたり、通常0.001mg/kg体重〜200mg/kg体重であり、好ましくは0.005mg/kg体重〜100mg/kg体重であり、より好ましくは0.01mg/kg体重〜50mg/kg体重である。 The ingestion amount or dose of the agent of the present invention refers to the muscle condition or the degree of muscle deterioration, sex, age in a subject to which the agent of the present invention is applied (hereinafter, also referred to as “application target” in the present specification). , Weight, dosage form, administration method, etc., but when the application target is a human adult, the amount converted to fucoxanthin is usually 0.001 mg/kg body weight to 200 mg/kg body weight per day. Yes, preferably 0.005 mg/kg body weight to 100 mg/kg body weight, more preferably 0.01 mg/kg body weight to 50 mg/kg body weight.
上記の1日あたりの摂取量または投与量は、1回で摂取させまたは投与してもよく、あるいは2回以上(たとえば、2〜5回)に分けて摂取させまたは投与することもできる。 The above-mentioned daily intake or dose may be ingested or administered once, or may be ingested or administered in two or more times (for example, 2 to 5 times).
本発明の剤の摂取または投与の時期は特に制限されず、食事の前または後、あるいは食事とともに摂取させまたは投与することができ、運動を実施させる場合は、運動開始前、運動中、運動終了後等のいずれの時期に摂取させまたは投与してもよい。
本発明の剤の摂取または投与の回数は特に制限されないが、筋質の向上を要する時に、少なくとも1回(1回または2回以上)である。
本発明の剤を摂取させまたは投与する回数が2回以上である場合、本発明の剤を摂取させまたは投与する期間(最初の摂取または投与から、最後の摂取または投与までの期間)は特に限定されないが、通常6時間〜4週間であり、より本発明の効果が発揮される観点から、好ましくは1日間〜2週間であり、より好ましくは3日間〜1週間である。
なお、本発明の剤に含有されるフコキサンチン等は、食経験の豊かな成分であり、安全性が高いため、本発明の剤は、連続して摂取させまたは投与することもできる。特に、筋質の低下を予防するためには、長期間(たとえば2週間以上)摂取させ、または投与することが好ましい。
The timing of ingestion or administration of the agent of the present invention is not particularly limited, and the agent can be ingested or administered before or after a meal, or together with a meal, and when exercise is performed, before exercise is started, during exercise, and after exercise is completed. It may be ingested or administered at any time such as after.
The number of times of ingestion or administration of the agent of the present invention is not particularly limited, but it is at least once (once or twice or more) when improvement of muscle quality is required.
When the agent of the present invention is ingested or administered more than once, the period of ingesting or administering the agent of the present invention (the period from the first ingestion or administration to the last ingestion or administration) is particularly limited. However, it is usually 6 hours to 4 weeks, preferably 1 day to 2 weeks, more preferably 3 days to 1 week, from the viewpoint of exerting the effect of the present invention.
Since the fucoxanthin and the like contained in the agent of the present invention are ingredients that have abundant eating experience and are highly safe, the agent of the present invention can be continuously ingested or administered. In particular, in order to prevent deterioration of muscle quality, it is preferable to ingest or administer for a long period (for example, 2 weeks or more).
本発明の剤は、単位包装形態とすることができる。本明細書において「単位包装形態」とは、特定量(たとえば、1回あたりの摂取量等)を1単位とし、該1単位または2単位以上が一つの容器または包装体に収容された形態をいい、たとえば、1回あたりの摂取量を1単位とする単位包装形態は、「1回あたりの摂取量単位の包装形態」と称する。単位包装形態に用いられる容器または包装体は、本発明の剤の形態等に応じて適宜選択し得るが、たとえば、紙製の容器または袋体、プラスチック製の容器または袋体、パウチ、アルミ缶、スチール缶、ガラス瓶、ペットボトル、PTP(press through pack)包装シート等が挙げられる。 The agent of the present invention can be in a unit packaging form. In the present specification, the term "unit packaging form" refers to a form in which a specific amount (for example, an intake amount per dose) is taken as one unit, and one unit or two or more units are contained in one container or package. That is, for example, a unit packaging form in which an intake amount per unit is 1 unit is referred to as a “ingestion unit amount packaging form”. The container or package used in the unit packaging form may be appropriately selected according to the form of the agent of the present invention, and examples thereof include a paper container or bag, a plastic container or bag, a pouch, and an aluminum can. , Steel cans, glass bottles, PET bottles, PTP (press through pack) packaging sheets, and the like.
本発明の剤の適用対象としては、哺乳動物(ヒト、マウス、ラット、ハムスター、ウサギ、ネコ、イヌ、ウシ、ウマ、ロバ、ブタ、ヒツジ、サル等)や、鳥類(ニワトリ等)が挙げられる。なお、本発明の剤をヒト以外の適用対象動物(以下、単に「対象動物」ともいう)に適用する場合、本発明の剤の摂取量または投与量は、対象動物の種類、性別、体重等に応じて適宜設定すればよい。 The application target of the agent of the present invention includes mammals (human, mouse, rat, hamster, rabbit, cat, dog, cow, horse, donkey, pig, sheep, monkey, etc.) and birds (chicken, etc.). .. When the agent of the present invention is applied to target animals other than humans (hereinafter, also simply referred to as “target animal”), the intake amount or dose of the agent of the present invention is the type, sex, weight, etc. of the target animal. It may be appropriately set according to
本発明の剤は、運動が制限される場合であっても、加齢等、種々の原因による筋質の低下を予防し、また、筋質を向上させることができる。さらに、無理のない程度の運動であっても、その効果を有効に増強させることができる。
従って、本発明の剤は、加齢により身体機能が低下した高齢者、疾患、負傷等により運動が制限される者等、従来筋質を向上させるのに必要とされていた運動を実施することが困難な者において、筋質の低下の予防や筋質の向上に好ましく用いられる。
The agent of the present invention can prevent deterioration of muscle quality due to various causes such as aging and improve muscle quality even when exercise is restricted. Furthermore, the effect can be effectively enhanced even if the exercise is reasonably done.
Therefore, the agent of the present invention is capable of performing exercise that has been conventionally required to improve muscle quality, such as the elderly who have deteriorated physical function due to aging, those whose movement is restricted due to illness, injury, etc. It is preferably used for prevention of deterioration of muscle quality and improvement of muscle quality in persons with difficulty
また、本発明の剤は、そのまま、またはさらに上記した賦形剤、溶剤、希釈剤等の添加剤を加えて、筋質向上用の医薬(以下、本明細書において「本発明の医薬」とも称する)とすることができる。
本発明の医薬におけるフコキサンチン等の含有量は、フコキサンチンに換算した量で通常0.0001重量%〜100重量%であり、好ましくは0.0005重量%〜100重量%であり、より好ましくは0.001重量%〜100重量%である。
本発明の医薬の投与量は、本発明の医薬が投与される患者の筋質低下の状態および程度、患者の年齢、性別、体重等により適宜定めることができるが、フコキサンチン等の投与量が、上記した1日あたりの投与量となるように設定することができる。
In addition, the agent of the present invention may be used as it is, or may be further added with additives such as the above-mentioned excipients, solvents, diluents, etc. to improve a muscle-improving drug (hereinafter, also referred to as “the drug of the present invention” in this specification). Can be referred to as).
The content of fucoxanthin or the like in the medicament of the present invention is usually 0.0001% by weight to 100% by weight in terms of fucoxanthin, preferably 0.0005% by weight to 100% by weight, and more preferably It is 0.001% by weight to 100% by weight.
The dose of the drug of the present invention can be appropriately determined depending on the state and degree of hypomuscularity of the patient to whom the drug of the present invention is administered, age, sex, weight of the patient, etc., but the dose of fucoxanthin or the like is The dose can be set to the above-mentioned daily dose.
本発明の医薬は、第十七改正日本薬局方製剤総則[3]製剤各条に記載された方法等、製剤の分野で周知の製剤化手段により、製造することができる。 The medicament of the present invention can be produced by a formulation means that is well known in the field of formulation, such as the method described in the 17th revised Japanese Pharmacopoeia General Provisions [3] Preparation Articles.
本発明の医薬は、高齢者、患者、要介護者等において、筋質の低下が認められ、または筋質の低下のおそれのある者に好適に投与され得る。 The medicament of the present invention can be suitably administered to elderly persons, patients, persons requiring nursing care, etc. who have muscle weakness or are at risk of muscle weakness.
さらに、本発明の剤は、各種食品に添加して使用することができる。本発明の剤が添加される食品は特に制限されず、一般的に食事やデザート等に供される形態の食品であれば如何なるものでもよい。たとえば、本発明の剤を飲料に添加し、所望により適当な風味を加えて、ドリンク剤(たとえば、清涼飲料等)とすることができる。より具体的には、本発明の剤は、たとえば、ジュース、牛乳、菓子、ゼリー、ヨーグルト、飴等に添加することができる。 Furthermore, the agent of the present invention can be used by adding it to various foods. The food to which the agent of the present invention is added is not particularly limited, and may be any food as long as it is generally used for meals, desserts and the like. For example, the agent of the present invention can be added to a beverage and, if desired, an appropriate flavor can be added to obtain a drink agent (for example, a soft drink). More specifically, the agent of the present invention can be added to, for example, juice, milk, confectionery, jelly, yogurt, candy and the like.
本発明の剤は、1日に摂取される量の食品に対し、フコキサンチン等の摂取量が、上記した1日あたりの摂取量となるように添加され得る。 The agent of the present invention can be added so that the intake amount of fucoxanthin or the like becomes the above-mentioned intake amount per day with respect to the daily intake amount of food.
本発明はまた、本発明の剤を含有する、筋質向上用の食品(以下、「本発明の食品」ともいう)を提供する。
本発明の食品は、本発明の剤と、必要に応じて、製造用剤、増粘安定剤、ガムベース、乳化剤、保存料、酸化防止剤、光沢剤、pH調整剤、甘味料、苦味料、酸味料、着色料、香料等の食品添加物を含有し、あるいは、本発明の剤と、食品もしくは食品原材料とを含有し、たとえばジュース類、清涼飲料水、茶類等の飲料;乳酸菌飲料、発酵乳、バター、チーズ、ヨーグルト、加工乳、脱脂乳等の乳製品;ハム、ソーセージ、ハンバーグ等の畜肉製品;蒲鉾、竹輪、さつま揚げ等の魚肉練り製品;だし巻き、卵豆腐等の卵製品;クッキー、ゼリー、チューイングガム、キャンディー、スナック菓子、冷菓等の菓子類;その他パン類、麺類、漬物類、燻製品、干物、佃煮、塩蔵品、スープ類、調味料等、種々の形態で提供することができ、瓶詰め食品、缶詰食品、レトルトパウチ食品とすることもできる。また、粉末状、顆粒状、シート状、カプセル状、タブレット状、ゼリー状等の形態とすることもできる。
The present invention also provides a food for improving muscle quality (hereinafter, also referred to as "food of the present invention") containing the agent of the present invention.
The food of the present invention, together with the agent of the present invention, if necessary, a manufacturing agent, a thickening stabilizer, a gum base, an emulsifier, a preservative, an antioxidant, a brightener, a pH adjuster, a sweetener, a bittering agent, Contains food additives such as acidulants, colorants, and flavors, or contains the agent of the present invention and foods or food raw materials, for example, beverages such as juices, soft drinks, teas; lactic acid bacteria beverages, Fermented milk, butter, cheese, yogurt, processed milk, skimmed milk, and other dairy products; ham, sausage, hamburgers, and other meat products; kamaboko, bamboo rings, fried fish meat, and other fish products; egg products such as dashi rolls and egg tofu; cookies , Jelly, chewing gum, candy, snacks, frozen desserts, etc.; other breads, noodles, pickles, smoked products, dried foods, boiled soup, salted foods, soups, seasonings, etc. It can also be bottled food, canned food, retort pouch food. Further, it may be in the form of powder, granules, sheet, capsule, tablet, jelly or the like.
本発明の食品は、筋質の低下のおそれのある、または筋質の向上の求められる高齢者、要介護者、患者等に好適に摂取させ得る。
また、本発明の食品は、高齢者ではないが筋質の低下の予防や筋質の向上を望む中・壮年者、介護は要しないが、疾患、負傷等のために運動の実施が制限される者等、筋質の向上を希望する者に幅広く摂取させることができる。
The food of the present invention can be suitably ingested by an elderly person, a care recipient, a patient, or the like who is likely to deteriorate muscle quality or is required to improve muscle quality.
Further, the food of the present invention is not an elderly person, but a middle-aged or middle-aged person who wants to prevent muscle deterioration or improve muscle quality, does not require care, but is restricted from performing exercise due to illness, injury, etc. It can be widely ingested by people who want to improve muscle quality, such as those who are sick.
従って、本発明の食品は、筋質の低下の予防または筋質の向上用の特定保健用食品、栄養機能食品、機能性表示食品等の保健機能食品、病者用食品、高齢者用食品等の特別用途食品、健康補助食品等としても提供され得る。 Therefore, the food of the present invention, food for specific health for prevention of muscle deterioration or improvement of muscle quality, food with nutritional function, food with health claims such as functionally labeled food, food for patients, food for elderly, etc. Can also be provided as a special-purpose food, health supplement, and the like.
さらに、本発明の剤を、濃厚流動食や、食品補助剤に添加して使用することも可能である。
「濃厚流動食」とは、1kcal/mL程度の濃度に調整され、長期間これのみを摂取する場合であっても、著しい栄養素の過不足が生じないように、各栄養素の質的構成が十分に考慮され、1日の栄養所要量をもとに設計された総合栄養食品(液体状食品)である。
本発明における「食品補助剤」とは、食品として摂取されるもの以外に栄養を補助する目的で摂取されるものをいい、栄養補助剤、サプリメントなどもこれに含まれる。本発明の剤を食品補助剤に添加する場合、所望により他の栄養成分や添加剤を加えて、例えば錠剤、カプセル、散剤、顆粒、懸濁剤、チュアブル剤、シロップ剤等の形態に調製することができる。
Furthermore, the agent of the present invention can be used by adding it to a concentrated liquid food or a food supplement.
The "concentrated liquid diet" is adjusted to a concentration of about 1 kcal/mL, and the qualitative composition of each nutrient is sufficient to prevent significant nutrient deficiency even when ingesting only this for a long time. Is a comprehensive nutritional food (liquid food) designed based on the daily nutritional requirement.
The “food supplement” in the present invention refers to those ingested for the purpose of supplementing nutrition, in addition to those ingested as food, and includes nutritional supplements and supplements. When the agent of the present invention is added to a food supplement, if desired, other nutritional ingredients or additives are added to prepare a tablet, capsule, powder, granule, suspension, chewable agent, syrup or the like. be able to.
上記した本発明の食品は、本発明の剤に、必要に応じて食品添加物を加えて、あるいは、食品または食品原材料中に本発明の剤を添加し、一般的な食品の製造方法を用いることにより、加工し、製造することができる。 The above-mentioned food of the present invention, to the agent of the present invention, optionally by adding a food additive, or by adding the agent of the present invention in the food or food raw materials, using a general method for producing foods By doing so, it can be processed and manufactured.
本発明の食品におけるフコキサンチン等の含有量は、食品の種類もしくは形態、当該食品の摂取により期待される筋質向上効果の程度等に応じて適宜設定され得るが、食品の総重量に対し、フコキサンチンに換算した量として、通常0.0001重量%〜100重量%程度であり、好ましくは0.0005重量%〜100重量%程度であり、より好ましくは0.001重量%〜100重量%程度である。 The content of fucoxanthin or the like in the food of the present invention may be appropriately set depending on the kind or form of the food, the degree of muscle improving effect expected by ingestion of the food, etc., with respect to the total weight of the food, The amount converted to fucoxanthin is usually about 0.0001% to 100% by weight, preferably about 0.0005% to 100% by weight, and more preferably about 0.001% to 100% by weight. Is.
また、本発明の食品の1日あたりの摂取量は、本発明の剤について上述した1日あたりの摂取量のフコキサンチン等を摂取できる量として設定され得る。 Further, the daily intake of the food of the present invention can be set as an amount at which the above-mentioned daily intake of fucoxanthin and the like for the agent of the present invention can be ingested.
本発明はまた、本発明の剤、並びに、本発明の剤を筋質の向上に使用することができること、または使用すべきであることを記載した記載物を含む、商業的パッケージをも提供する。 The present invention also provides a commercial package comprising the agent of the present invention and a description that describes that the agent of the present invention can be or should be used for improving muscle quality. ..
さらに本発明は、筋質を向上させる必要のある対象動物の筋質の向上方法(以下、本明細書において「本発明の方法」ともいう)をも提供する。 Furthermore, the present invention also provides a method for improving muscle quality of a target animal that requires improvement in muscle quality (hereinafter, also referred to as “method of the present invention” in the present specification).
本発明の方法は、筋質を向上させる必要のある対象動物に、当該対象動物の筋質を向上させるのに有効な量のフコキサンチン等を摂取させること、または投与することを含む。 The method of the present invention includes ingesting or administering to a target animal in need of improving muscle quality an amount of fucoxanthin or the like effective for improving the muscle quality of the target animal.
本発明の方法における対象動物としては、ヒトおよび、マウス、ラット、ハムスター、ウサギ、ネコ、イヌ、ウシ、ウマ、ロバ、ブタ、ヒツジ、サル等のヒト以外の哺乳動物、ならびにニワトリなどの鳥類が挙げられる。
ヒトの場合、本発明の方法は、筋質の低下が認められる者や、筋質の低下の予防または筋質の向上を望む者に対して幅広く適用することができるが、特に身体機能の低下の認められる高齢者や要介護者、疾患、負傷等により運動が制限される者等、十分な運動を実施することのできない患者等に好適に適用される。
本発明の方法におけるフコキサンチン等の摂取量または投与量は、対象動物の種類、年齢、体重、筋質の低下の状態、程度等に応じて適宜設定されるが、本発明の剤において、ヒトおよびヒト以外の対象動物について、上記した摂取量または投与量と同様の量を、上記した回数および期間にて摂取させまたは投与することができる。
さらに、本発明の方法におけるフコキサンチン等の摂取または投与の方法としては、経口摂取または経口投与、経腸経管投与、輸液による投与等が挙げられるが、医療機関にて医師の指導監督下に行う必要がなく、簡便に行うことができることから、経口摂取または経口投与が好ましい。
The target animals in the method of the present invention, humans and, non-human mammals such as mice, rats, hamsters, rabbits, cats, dogs, cows, horses, donkeys, pigs, sheep, monkeys, and birds such as chickens. Can be mentioned.
In the case of humans, the method of the present invention can be widely applied to those who have muscle weakness and those who want to prevent muscle weakness or improve muscle quality, but especially to decrease physical function. It is preferably applied to patients who cannot exercise sufficiently, such as the elderly who are admitted, those requiring care, those whose exercise is restricted due to illness, injury, and the like.
The intake or dose of fucoxanthin or the like in the method of the present invention is appropriately set depending on the type of the target animal, age, weight, state of muscle deterioration, degree, etc., but in the agent of the present invention, human Also, with respect to non-human target animals, the same amount as the above-mentioned intake amount or dose can be ingested or administered at the above-mentioned number of times and period.
Further, the method of ingestion or administration of fucoxanthin or the like in the method of the present invention, orally ingested or oral administration, enteral tube administration, administration by infusion, etc., under the supervision of a doctor at a medical institution. Oral ingestion or oral administration is preferable because it can be easily performed without needing to perform it.
以下、試験例により本発明をさらに詳細に説明する。 Hereinafter, the present invention will be described in more detail with reference to test examples.
[試験例1]筋インスリンシグナル低下に対するフコキサンチン誘導体の作用の検討
フコキサンチン誘導体として、フコキサンチンの代謝生成物であるフコキサチノールを用い、筋同化シグナルをインスリンシグナルでモニターし、そのインスリンシグナル低下に対するフコキサンチノールの作用を評価した。
評価は、下記の通り、マウス由来の筋細胞株C2C12に飽和脂肪酸を暴露させることにより、インスリンシグナル低下をもたらす評価系を用いて実施した。
[Test Example 1] Investigation of action of fucoxanthin derivative on reduction of muscle insulin signal By using fucoxanthin, which is a metabolite of fucoxanthin, as a fucoxanthin derivative, muscle assimilation signal was monitored by insulin signal, and the action on the reduction of insulin signal was detected. The effect of coxanthinol was evaluated.
The evaluation was performed using an evaluation system that causes a decrease in insulin signal by exposing the mouse-derived muscle cell line C2C12 to saturated fatty acids, as described below.
(1)C2C12筋管細胞(Myotubes)への分化誘導
マウスC3H 骨格筋筋芽細胞(muscle myoblast)(C2C12)細胞(DSファーマバイオメデカル株式会社)を用いて、本評価を実施した。
筋管細胞(Myotubes)への分化誘導は、筋芽細胞(Myoblasts)の継代数が12〜18である細胞を用いて行った。筋芽細胞(Myoblasts)を0.3×105〜0.5×105/wellの細胞数で12ウェル(直径=3cm)プレートに蒔種し、細胞増殖の状態が90%コンフルーエントになったのを確認した後、2(w/v)% ウマ血清 (HS)、1(w/v)% ペニシリン−ストレプトマイシン−グルタミンを含有するDMEM培養液(2(w/v)%HS、1(w/v)%P/S、1(w/v)%Gln/DMEM)に交換し、筋管細胞(Myotubes)への分化を誘導した。筋管細胞(Myotubes)への分化を確認した後、筋インスリンシグナル評価系の構築、および構築した評価系を用いてフコキサンチノールの作用の評価を行った。
(1) Induction of differentiation into C2C12 myotubes (Myotubes) This evaluation was carried out using mouse C3H skeletal myoblast (C2C12) cells (DS Pharma Biomedical Co., Ltd.).
Induction of differentiation into myotubes (Myotubes) was performed using cells having a passage number of 12 to 18 for myoblasts. Myoblasts (Myoblasts) were seeded in a 12-well (diameter = 3 cm) plate at a cell number of 0.3 × 10 5 to 0.5 × 10 5 /well, and it was confirmed that the cell growth state became 90% confluent. After that, DMEM culture solution containing 2 (w/v)% horse serum (HS), 1 (w/v)% penicillin-streptomycin-glutamine (2 (w/v)% HS, 1 (w/v)) % P/S, 1 (w/v)% Gln/DMEM) to induce differentiation into myotubes (Myotubes). After confirming differentiation into myotubes (Myotubes), a muscle insulin signal evaluation system was constructed, and the action of fucoxanthinol was evaluated using the constructed evaluation system.
(2)筋インスリンシグナルの評価
筋インスリンシグナルの評価の概要を図1に示した。
筋管細胞(Myotubes)を用いて、0.5mMのパルミチン酸(FFA)/0.5(w/v)%アルブミン(BSA)/DMEM培養液中で一晩(17時間〜24時間)、37℃にて、5(v/v)%二酸化炭素存在下で培養した。
一晩培養後、DMEM(BSA/FFA無添加)で約2時間プレインキュベーションし、100nMのインスリンによりインキュベーション(15分間〜20分間)して刺激した後、筋管細胞を回収した(各n=3)。回収した細胞から、RIPA(Radio-Immunoprecipitation Assay)バッファー(セルシグナリングテクノロジーズ社(Cell Signaling Technologies Inc.))によりタンパク質を抽出し、タンパク質の最終濃度が0.5mg/mLになるように調整し、試料とした。
上記試料について、リン酸化Akt(pAkt)(Ser473)(# 9271S;セルシグナリングテクノロジーズ社(Cell Signaling Technologies Inc.))、および全Akt(total Akt)(#9272S;セルシグナリングテクノロジーズ社(Cell Signaling Technologies Inc.))の発現量を、ウエスタンブロッティング(WB)法で定量した。各タンパク質の発現量はウェスタンブロット−化学発光イメージングシステム(フージョン(Fusion)FX)(ヴィルバー−ルーマット社(Vilber-Lourmat Inc.))を用いて定量した。
各濃度のフコキサンチンはFFAと同時に添加し、筋インスリンシグナルの回復を指標に、筋質向上効果を検討した。フコキサンチノールの最終濃度は1μM、3μMおよび10μMになるように調整した。また、ポジティブコントロールとして、抗酸化活性物質であるN−アセチルシステイン(NAC)5mMを用いた。
(2) Evaluation of muscle insulin signal The outline of evaluation of muscle insulin signal is shown in FIG.
Using myotubes (Myotubes), 0.5 mM palmitic acid (FFA)/0.5 (w/v)% albumin (BSA)/DMEM medium overnight (17 to 24 hours), 37 It was cultured in the presence of 5 (v/v)% carbon dioxide at 0°C.
After culturing overnight, preincubation with DMEM (without BSA/FFA) for about 2 hours and incubation with 100 nM insulin (15 to 20 minutes) for stimulation were performed, and then myotube cells were collected (each n=3). ). A protein is extracted from the collected cells with a RIPA (Radio-Immunoprecipitation Assay) buffer (Cell Signaling Technologies Inc.), and the final concentration of the protein is adjusted to 0.5 mg/mL, and a sample is prepared. And
Regarding the above sample, phosphorylated Akt (pAkt) (Ser473) (# 9271S; Cell Signaling Technologies Inc.), and total Akt (total Akt) (#9272S; Cell Signaling Technologies Inc. .)) was quantified by Western blotting (WB) method. The expression level of each protein was quantified using a Western blot-chemiluminescence imaging system (Fusion FX) (Vilber-Lourmat Inc.).
Fucoxanthin at each concentration was added at the same time as FFA, and the effect of improving muscle quality was examined using the recovery of muscle insulin signal as an index. The final concentration of fucoxanthinol was adjusted to be 1 μM, 3 μM and 10 μM. As a positive control, 5 mM of N-acetyl cysteine (NAC), which is an antioxidant active substance, was used.
(3)評価結果
リン酸化Akt(Ser473)および全Aktの発現量の測定結果から、インスリン刺激のない場合のリン酸化Aktと全Aktの比に対するインスリン刺激時のリン酸化Aktと全Aktの比[(pAkt/Total Akt) Insulin+]/[(pAkt/Total Akt) Insulin-]を求め、リン酸化指数(stimulation index)として図2に示した。図2中、リン酸化指数は、各試料について、平均値±標準誤差にて示した。
図2において、BSAを添加して培養した筋管細胞において、インスリン刺激によってAktのリン酸化の上昇が認められた。0.5mMのFFAを添加して培養した細胞では、インスリン刺激によるAktのリン酸化作用は抑制されており、筋インスリンシグナルの低下が示唆された。
一方、フコキサンチノールは、FFAによる筋インスリンシグナルの低下を、用量依存的に回復させることが認められた。また、フコキサンチノールは、NACより低濃度で、NACと同等以上の作用を示すことが認められた。
本試験例の結果から、フコキサンチンが筋インスリンシグナルの上昇をもたらす成分として有用である可能性が示唆された。
(3) Evaluation results From the results of measuring the expression levels of phosphorylated Akt (Ser473) and total Akt, the ratio of phosphorylated Akt and total Akt during insulin stimulation to the ratio of phosphorylated Akt and total Akt in the absence of insulin stimulation [ (pAkt/Total Akt) Insulin+]/[(pAkt/Total Akt) Insulin-] was calculated and shown as a phosphorylation index (stimulation index) in FIG. In FIG. 2, the phosphorylation index is shown as the average value±standard error of each sample.
In Fig. 2, in myotube cells cultured with the addition of BSA, an increase in Akt phosphorylation due to insulin stimulation was observed. In cells cultured with the addition of 0.5 mM FFA, the phosphorylation effect of Akt upon insulin stimulation was suppressed, suggesting a decrease in muscle insulin signal.
On the other hand, fucoxanthinol was found to restore the FFA-induced decrease in muscle insulin signal in a dose-dependent manner. Further, it was confirmed that fucoxanthinol exhibits an action equal to or higher than NAC at a lower concentration than NAC.
From the results of this test example, it was suggested that fucoxanthin may be useful as a component that causes an increase in muscle insulin signal.
[試験例2]フコキサンチン誘導体の筋ミトコンドリア機能に対する作用の検討
フコキサンチン誘導体として、フコキサンチンの代謝生成物であるフコキサチノールを用い、筋ミトコンドリアの機能低下に対する作用を評価した。
評価は、マウス由来の筋細胞株C2C12を用い、下記の通り、飽和脂肪酸を暴露させることにより、骨格筋の質の変化として観察される筋ミトコンドリア機能の低下を、酸素消費量を指標として観察することにより実施した。細胞外酸素消費量は、細胞外フラックスアナライザーXFe24 (プライムテック株式会社)を用いて測定した。
[Test Example 2] Examination of Action of Fucoxanthin Derivative on Muscle Mitochondrial Function Fucoxanthin, which is a metabolite of fucoxanthin, was used as a fucoxanthin derivative to evaluate the action on muscle mitochondrial function decline.
The evaluation was performed using a mouse-derived muscle cell line C2C12, and by observing a decrease in muscle mitochondrial function, which is observed as a change in quality of skeletal muscle, by exposing saturated fatty acids, as described below, using oxygen consumption as an index. It was carried out by The extracellular oxygen consumption was measured using an extracellular flux analyzer XFe24 (Primetech Co., Ltd.).
細胞外フラックスアナライザーXFe24測定用プレート(FluxPak−XFe24 アッセイパック/PS:102340-100)に、C2C12筋芽細胞(myoblast)(1.8×104 cells/well/100μL)を播種し、3〜4日後に分化培地(2(w/v)%HS、1(w/v)%P/S、1(w/v)%GlutaMAX(商標)−I(L−アラニル−L−グルタミン)(35050-061、gibco)、0.3mM HEPES(4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid)/DMEM)に交換して培養し、筋管細胞(Myotubes)に分化したことを確認した後、0.5mMのパルミチン酸(FFA)/0.5(w/v)%BSA/DMEM培養液中でさらに一晩(17時間〜24時間)培養し、筋ミトコンドリア機能の評価を実施した。
フコキサンチノールは、FFAと同時に、最終濃度が100nMとなるように添加した。
フラックスアナライザーXFe24における筋ミトコンドリア機能の評価は、プライムテック社より提唱されている測定方法に準じて、3.0μMのオリゴマイシン(ATP合成酵素阻害剤;75351、シグマ(Sigma)社)、3.0μMのカルボニルシアニド−p−トリフルオロメトキシフェニルヒドラゾン(FCCP)(脱共役剤;C2920、シグマ(Sigma)社)、0.5μMのアンチマイシンA(ミトコンドリア複合体III阻害剤;A8674、シグマ(Sigma)社)、0.5μMのロテノン(ミトコンドリア複合体I阻害剤;R8875、シグマ(Sigma)社)を順次添加し、酸素消費速度(OCR)を測定して行った。
Extracellular flux analyzer XFe24 measurement plate (FluxPak-XFe24 assay pack/PS: 102340-100) was seeded with C2C12 myoblasts (1.8×10 4 cells/well/100 μL) and 3 to 4 days later. Differentiation medium (2 (w/v)% HS, 1 (w/v)% P/S, 1 (w/v)% GlutaMAX™-I (L-alanyl-L-glutamine) (35050-061, gibco), 0.3 mM HEPES (4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid)/DMEM) was exchanged and cultured, and it was confirmed that the cells were differentiated into myotubes (Myotubes). The cells were further cultured overnight (17 to 24 hours) in a palmitic acid (FFA)/0.5 (w/v)% BSA/DMEM culture solution to evaluate muscle mitochondrial function.
Fucoxanthinol was added simultaneously with FFA to a final concentration of 100 nM.
The evaluation of the muscle mitochondrial function in the flux analyzer XFe24 was carried out according to the measurement method proposed by Primetech, in which 3.0 μM oligomycin (ATP synthase inhibitor; 75351, Sigma), 3.0 μM Carbonyl cyanide-p-trifluoromethoxyphenylhydrazone (FCCP) (uncoupler; C2920, Sigma), 0.5 μM antimycin A (mitochondrial complex III inhibitor; A8674, Sigma). , 0.5 μM rotenone (mitochondrial complex I inhibitor; R8875, Sigma) were sequentially added, and the oxygen consumption rate (OCR) was measured.
評価結果は、図3に示した。図3中、OCRは各試料について、平均値±標準誤差にて示した。
図3において、FCCP添加後のOCR値と、アンチマイシンAおよびロテノン添加後のOCR値から求められる筋ミトコンドリアの最大呼吸量は、0.5mMのFFAに暴露することにより低下した。一方、100nmのフコキサンチノールの添加により、低下した最大呼吸量の回復が認められた。
上記試験例の結果から、フコキサンチンは筋ミトコンドリア機能の低下を回復させる効果を有することが示唆された。
The evaluation results are shown in FIG. In FIG. 3, OCR is shown as the average value±standard error of each sample.
In FIG. 3, the maximum respiration of muscle mitochondria obtained from the OCR value after addition of FCCP and the OCR values after addition of antimycin A and rotenone was decreased by exposure to 0.5 mM FFA. On the other hand, the recovery of the lowered maximum respiratory volume was recognized by the addition of 100 nm of fucoxanthinol.
From the results of the above test examples, it was suggested that fucoxanthin has an effect of recovering the decrease in muscle mitochondrial function.
[試験例3]フコキサンチンの筋量および筋機能に対する作用の検討(in vivo評価)
フコキサンチンの摂取が、筋量および筋機能に及ぼす影響を、下記の通り評価した。
評価は、高脂肪食摂取によって筋量の低下や筋機能の低下を呈するモデル動物を用いて実施した。
6週齢の正常Sprague-Dawley(SD)雄性ラット(日本チャールス・リバー株式会社(神奈川)より購入)を馴化飼育し、9週齢の時点で表1に示す通り4群に分け、通常餌(Normal)および30重量%の脂肪を含む餌(HF)の各実験食の供与と、運動およびフコキサンチンの投与を開始した。フコキサンチンは、0.2重量%の含有量となるように餌に混ぜて供与した。
実験スケジュールを図4に示した。運動を行わせる群(HF/EX群およびHF/EX+Fco群)については、週に1回運動を実施した。
HF供与5週目の時点で、筋機能(筋力)の測定を行い、HF供与6週目に剖検し、筋重量を測定した。
[Test Example 3] Examination of action of fucoxanthin on muscle mass and muscle function (in vivo evaluation)
The effects of ingestion of fucoxanthin on muscle mass and muscle function were evaluated as follows.
The evaluation was carried out using a model animal that exhibits decreased muscle mass and decreased muscle function due to intake of a high-fat diet.
Six-week-old normal Sprague-Dawley (SD) male rats (purchased from Charles River Japan, Inc. (Kanagawa)) were acclimated and divided into 4 groups at 9 weeks of age as shown in Table 1, and fed with normal diet ( Normal) and a diet containing 30% by weight of fat (HF), and the administration of exercise and fucoxanthin were started. Fucoxanthin was supplied by being mixed with the feed so that the content of fucoxanthin was 0.2% by weight.
The experimental schedule is shown in FIG. The exercise groups (HF/EX group and HF/EX+Fco group) were exercised once a week.
Muscle function (muscular strength) was measured at 5 weeks after HF delivery, and autopsy was performed at 6 weeks after HF delivery, and muscle weight was measured.
運動は、ラットの左脚を用いて、局所のレジスタンス運動負荷を行って実施した。小動物用足関節運動装置(バイオリサーチセンター株式会社)に、イソフルラン麻酔下のラットを静置し、前脛骨筋に皮膚刺激電極を装着し、電気刺激により収縮させた。同時に前脛骨筋が収縮した方向とは反対方向へ牽引し、伸張負荷をかけた。運動負荷設定条件は以下の通りである。
(i)収縮(電気)負荷条件:4mA〜5mA、100Hz、1100msec
(ii)伸張負荷条件:左脚関節の角度を90°から135°へ速度=100deg/secにて伸張
(iii)運動負荷頻度および回数:10秒毎に10回を1セットとし、5セット繰り返して実施(合計50回)、各セットごとに60秒間の休憩をはさむ
The exercise was performed by using the left leg of the rat and performing local resistance exercise load. A rat under isoflurane anesthesia was placed in an ankle exercising device for small animals (Bioresearch Center Co., Ltd.), a skin stimulating electrode was attached to the tibialis anterior muscle, and contracted by electrical stimulation. At the same time, the tibialis anterior muscle was pulled in the direction opposite to the contracted direction, and an extension load was applied. The exercise load setting conditions are as follows.
(I) Contraction (electrical) load condition: 4 mA to 5 mA, 100 Hz, 1100 msec
(Ii) Stretching load condition: The angle of the left leg joint is stretched from 90° to 135° at a speed of 100 deg/sec. (iii) Exercise load frequency and frequency: 10 times every 10 seconds, 1 set is repeated and 5 sets are repeated. (50 times in total) with a 60 second break between each set
筋力の測定は、小動物用足関節運動装置(バイオリサーチセンター株式会社)を用いて、イソフルラン麻酔下で行った。ラットの左脚関節と左膝関節の角度を90°に固定し、固定した左前脛骨筋上に皮膚刺激電極を装着した。装着後、2.2mAの電気刺激 (収縮刺激)に対する最大不随意筋力を筋力として算出した。 The muscle strength was measured under anesthesia with isoflurane using a small animal ankle joint exercising device (Bio Research Center Co., Ltd.). The angle between the left leg joint and the left knee joint of the rat was fixed at 90°, and a skin stimulation electrode was mounted on the fixed left tibialis anterior muscle. After wearing, the maximum involuntary muscular strength to an electric stimulus (contraction stimulus) of 2.2 mA was calculated as muscular strength.
筋量(前脛骨筋の重量)の測定結果を図5に、2.2mAの電気刺激 (収縮刺激)時の筋力の測定結果を図6に示した。各測定結果は、平均値±標準誤差にて示した。また、各測定結果について、Normal群およびHF/Vehicle群との間でチューキー−クラマー(Tukey-Kramaer)の検定を実施した。
図中、「*」は、Normal群との間にp<0.05にて有意差があることを示す。また、「++」および「+++」は、HF/Vehicle群との間にそれぞれp<0.01およびp<0.001にて有意差があることを示す。
The results of measurement of muscle mass (tibialis anterior muscle weight) are shown in FIG. 5, and the results of measurement of muscle strength during electrical stimulation (contraction stimulation) of 2.2 mA are shown in FIG. Each measurement result was shown by the average value +/- standard error. In addition, for each measurement result, Tukey-Kramaer test was performed between the Normal group and the HF/Vehicle group.
In the figure, “*” indicates that there is a significant difference from the Normal group at p<0.05. Moreover, "++" and "++++" indicate that there is a significant difference between the HF/Vehicle group and p<0.01 and p<0.001, respectively.
図5において、HF/Vehicle群の前脛骨筋(運動刺激部位)の相対的重量は、Normal群に対し、有意に低下したことが認められた。一方、運動を行った群(HF/EX群)では、HF/Vehicle群に比べて筋重量が有意に増加したが、運動に加えてフコキサンチンを摂取させた群(HF/EX+Fco群)では、筋重量がさらに増加したことが認められた。 In FIG. 5, it was confirmed that the relative weight of the tibialis anterior muscle (motion stimulation site) in the HF/Vehicle group was significantly decreased as compared with the Normal group. On the other hand, in the exercise group (HF/EX group), the muscle weight was significantly increased as compared with the HF/Vehicle group, but in the group that received fucoxanthin in addition to exercise (HF/EX+Fco group), It was observed that the muscle weight increased further.
図6において、HF/Vehicle群の筋力は、Normal群に比べて低下し、筋機能が低下した可能性が示唆された。高脂肪食摂取による前記筋力の低下は、運動を行ったHF/EX群で抑制される傾向が認められた。運動に加えてフコキサンチンを摂取させたHF/EX+Fco群では、高脂肪食の摂取により低下した筋力が有意に向上したことが認められ、運動に加えてフコキサンチンを摂取させることにより、低下した筋機能をより改善し得ることが示唆された。 In FIG. 6, the muscle strength of the HF/Vehicle group was lower than that of the Normal group, suggesting that the muscle function may be lowered. The decrease in muscle strength due to intake of a high-fat diet tended to be suppressed in the exercised HF/EX group. In the HF/EX+Fco group that received fucoxanthin in addition to exercise, it was found that the muscle strength decreased by the intake of a high-fat diet was significantly improved, and the muscle decreased by intake of fucoxanthin in addition to exercise. It was suggested that the function could be improved.
以上詳述したように、本発明により、運動が制限される場合であっても、加齢等、種々の原因による筋質の低下を予防し、また、筋質を向上させることができ、さらに、無理のない程度の運動であっても、その効果を有効に増強させることができる筋質向上剤を提供することができる。
本発明の筋質向上剤は、加齢による身体機能の低下や、疾患、負傷等により運動が制限される者等、従来筋質を向上させるのに必要とされていた運動を実施することが困難な者においても、筋質の低下の予防や筋質の向上に有効に利用され得る。
As described in detail above, according to the present invention, even when exercise is restricted, it is possible to prevent deterioration of muscle quality due to various causes such as aging, and also improve muscle quality, and Thus, it is possible to provide a muscular strength improving agent that can effectively enhance the effect of the exercise even if the exercise is not difficult.
The muscle-improving agent of the present invention can carry out the exercise conventionally required to improve the muscle quality, such as a person whose physical function is deteriorated due to aging, exercise is restricted due to diseases, injuries and the like. Even in difficult people, it can be effectively used for prevention of muscle deterioration and improvement of muscle quality.
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