WO2018066394A1 - Composition for making muscles flexible - Google Patents

Composition for making muscles flexible Download PDF

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WO2018066394A1
WO2018066394A1 PCT/JP2017/034436 JP2017034436W WO2018066394A1 WO 2018066394 A1 WO2018066394 A1 WO 2018066394A1 JP 2017034436 W JP2017034436 W JP 2017034436W WO 2018066394 A1 WO2018066394 A1 WO 2018066394A1
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fatty acid
acid
composition
muscle
fish oil
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PCT/JP2017/034436
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French (fr)
Japanese (ja)
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賢一 柳本
韓　力
二功 川端
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日本水産株式会社
国立大学法人 九州大学
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Priority to JP2018543844A priority Critical patent/JPWO2018066394A1/en
Publication of WO2018066394A1 publication Critical patent/WO2018066394A1/en

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    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/115—Fatty acids or derivatives thereof; Fats or oils
    • A23L33/12—Fatty acids or derivatives thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/185—Acids; Anhydrides, halides or salts thereof, e.g. sulfur acids, imidic, hydrazonic or hydroximic acids
    • A61K31/19—Carboxylic acids, e.g. valproic acid
    • A61K31/20—Carboxylic acids, e.g. valproic acid having a carboxyl group bound to a chain of seven or more carbon atoms, e.g. stearic, palmitic, arachidic acids
    • A61K31/202—Carboxylic acids, e.g. valproic acid having a carboxyl group bound to a chain of seven or more carbon atoms, e.g. stearic, palmitic, arachidic acids having three or more double bonds, e.g. linolenic
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/21—Esters, e.g. nitroglycerine, selenocyanates
    • A61K31/215—Esters, e.g. nitroglycerine, selenocyanates of carboxylic acids
    • A61K31/22—Esters, e.g. nitroglycerine, selenocyanates of carboxylic acids of acyclic acids, e.g. pravastatin
    • A61K31/23—Esters, e.g. nitroglycerine, selenocyanates of carboxylic acids of acyclic acids, e.g. pravastatin of acids having a carboxyl group bound to a chain of seven or more carbon atoms
    • A61K31/232—Esters, e.g. nitroglycerine, selenocyanates of carboxylic acids of acyclic acids, e.g. pravastatin of acids having a carboxyl group bound to a chain of seven or more carbon atoms having three or more double bonds, e.g. etretinate
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/66—Phosphorus compounds
    • A61K31/683—Diesters of a phosphorus acid with two hydroxy compounds, e.g. phosphatidylinositols
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/56—Materials from animals other than mammals
    • A61K35/60—Fish, e.g. seahorses; Fish eggs
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/56—Materials from animals other than mammals
    • A61K35/612—Crustaceans, e.g. crabs, lobsters, shrimps, krill or crayfish; Barnacles
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/66—Microorganisms or materials therefrom
    • A61K35/68—Protozoa, e.g. flagella, amoebas, sporozoans, plasmodium or toxoplasma
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P21/00—Drugs for disorders of the muscular or neuromuscular system

Definitions

  • the present invention relates to a composition for softening muscles.
  • it is a composition for ingestion in order to increase the flexibility of skeletal muscle, which is said to affect motor function.
  • Muscle flexibility is an important factor for health promotion and injury prevention, and it is considered that sports injury can be prevented and reduced by maintaining and improving muscle flexibility (Non-Patent Document 1).
  • muscle flexibility and sports performance are also related, and it is known that running economy is improved by improving muscle flexibility by stretching (Non-patent Document 1). Therefore, it is considered that improving muscle flexibility leads to prevention / improvement of injury and improvement of exercise performance (Non-Patent Document 1).
  • Non-patent Document 2 Fish oil is rich in polyunsaturated fatty acids (hereinafter also referred to as PUFA) such as eicosapentaenoic acid (hereinafter also referred to as EPA) and docosahexaenoic acid (hereinafter also referred to as DHA).
  • PUFA polyunsaturated fatty acids
  • EPA eicosapentaenoic acid
  • DHA docosahexaenoic acid
  • the present invention focuses on muscle flexibility, which is attracting attention in the field of sports and geriatrics, and supports muscle flexibility by exercising and massage, etc. It is an object of the present invention to provide a composition that enhances the viscosity.
  • PUFA having a low viscosity is taken into the erythrocyte membrane, so that red blood cells become flexible and blood flow is smooth even in narrow blood vessels.
  • ingestion of PUFA would bring PUFA into skeletal muscle and improve skeletal muscle flexibility.
  • the flexibility of the skeletal muscle of rats fed with fish oil for several weeks was measured using a muscle hardness meter and rheometer. Confirmed and completed the present invention.
  • the gist of the present invention is the following composition (1) to (8) for softening muscles by oral ingestion.
  • composition according to (1) containing n-3 polyunsaturated fatty acids as refined fish oil, refined krill oil, and refined microbial oil.
  • FIG. 1 It is a figure which shows the shear strength of the anterior tibial muscle of the rat extract
  • FIG. It is a figure which shows the muscle hardness change rate of the rat thigh in Example 2.
  • FIG. It is a figure which shows the correlation with the shear strength of a rectus femoris muscle in the rat extract
  • PUFA is a fatty acid having 18 or more carbon atoms and 3 or more double bonds, preferably a fatty acid having 20 or more carbon atoms and 3 or more double bonds, more preferably 20 or more carbon atoms and a double bond. It is a fatty acid having 4 or more bonds.
  • n-3 PUFA include ⁇ -linolenic acid, eicosapentaenoic acid, docosapentaenoic acid, and docosahexaenoic acid.
  • the PUFA used in the present invention can be used in the form of glycerides or phospholipids containing free fatty acids, lower alcohol esters, or n-3 polyunsaturated fatty acids as constituent fatty acids.
  • esters with glycerin i.e. triglycerides, diglycerides, monoglycerides, or phospholipids, i.e. Or an ester with a lower alcohol (having 1 to 3 carbon atoms), that is, a methyl ester, an ethyl ester, or the like.
  • Preference is given to those containing n-3 fatty acids as the main component, such as EPA and DHA, and substantially free of n-6 fatty acids.
  • the total of n-6 fatty acids is 10% by weight or less, preferably 5% by weight or less.
  • triglycerides refined fish oil, refined microbial oil, or concentrated oil with a higher concentration of n-3 PUFA contained in them (enzyme reaction, chemical reaction to increase the concentration of n-3 PUFA in refined fish oil) Concentrated) can be used.
  • refined krill oil, refined microbial oil, etc. can be used as a phospholipid.
  • N-3 PUFA sources containing EPA and DHA include fish and shellfish extracts, animal extracts, egg yolk extracts, plant extracts, fungal extracts, etc., preferably krill oil, fish oil, squid extract, skipjack ovary extract Products, extracts of genetically modified plants, extracts of Labyrinthula, and the like.
  • Examples of the material containing a large amount of phospholipid include krill oil and squid extract.
  • the phospholipid concentration and purity in these materials can be arbitrarily adjusted by using techniques generally known in the art such as concentration, extraction, and / or purification, and blending.
  • EPA and / or DHA are known to be effective for hyperlipidemia and allergic symptoms, and many pharmaceuticals and various supplements are already on the market.
  • capsule preparations similar to these can be used. Specifically, a product obtained by processing a fat and oil obtained by adding an antioxidant to purified fish oil into a gelatin capsule is often used.
  • the dosage form of the preparation is not particularly limited as long as it is an orally administrable dosage form, and is a tablet, capsule (hard capsule, soft capsule), microcapsule, granule, fine granule, powder, oral liquid preparation (emulsion, Suspensions, etc.), syrups, jellies and the like.
  • a dosage form enclosed in a capsule particularly a soft capsule or a microcapsule is preferable.
  • Each formulation is prepared by methods known in the art, and usually used excipients, diluents, binders, wetting agents, disintegrants, disintegration inhibitors, absorption enhancers, lubricants, solubilizers, Prepared using buffers, emulsifiers, suspending agents and the like.
  • Additives include stabilizers, preservatives, buffers, isotonic agents, chelating agents, pH adjusters, surfactants, colorants, flavoring agents, flavoring agents, sweetening agents that are usually used depending on the dosage form. An agent or the like can be used.
  • the amount of n-3 PUFA contained in the preparation is, for example, 20% by weight or more, preferably 40% by weight or more, or 50% by weight or more, more preferably 80% by weight or more as the total amount of n-3 PUFA. is there.
  • any fish oil may be used as long as it contains EPA and DHA.
  • fish oils rich in EPA include sardine oil and cod liver oil
  • fish oils rich in DHA include tuna oil and bonito oil.
  • the concentrated oil of PUFA is a fat and oil in which the concentration of EPA / DHA in the triglyceride of refined fish oil is increased by using a lipase having regioselectivity and fatty acid selectivity.
  • n-3 PUFAs such as EPA and DHA are preferably taken orally in a total amount of 5 to 150 mg / kg per day. More preferably, it is 15 to 150 mg / kg, and particularly preferably 15 to 75 mg / kg. It may be taken at once or divided. It is preferable to ingest continuously, and the effect is recognized by continuous administration for at least 2 weeks.
  • n-3 PUFA is a food-derived ingredient, and since it has already been used for a long time in other applications, it can be safely consumed.
  • the use target is not particularly limited, and it can be used for subjects who want to soften muscles, from young people to elderly people. It is particularly preferable for those who perform sports or those who are concerned about muscle hardening due to age. It can also be applied to mammalian animals as objects of use. It is particularly preferable to use it for competition animals such as horses and dogs.
  • the composition of the present invention can be ingested as a food or together with a food ingredient because the active ingredient is extremely safe. Examples of the present invention will be described below, but the present invention is not limited thereto.
  • ⁇ Experiment method> 1 Experimental animals 7-week-old Wistar male rats were used as animals. The experiment was conducted after one week of pre-breeding. Rats were divided into 2 groups (6 groups), EPA / DHA-rich fish oil-added feed intake group (fish oil group) and lard-added feed intake group (Lard group). The feed was pair-feeded and the food intake was adjusted. Water was ad libitum. The animals were housed in individual cages made of stainless steel in a conditioned animal room with a room temperature of 22 ⁇ 2 ° C., 50 ⁇ 10% humidity, and illumination for 12 hours. 2.
  • Table 1 shows the fatty acid composition of the fish oil and lard used
  • Table 2 shows the composition of the AIN-93G modified feed.
  • the feeding period was 4 weeks. After the breeding period, the anterior tibial muscle (TA) was collected and stored frozen at ⁇ 80 ° C. until analysis.
  • TA anterior tibial muscle
  • Shear titration measurement The weight of the anterior tibial muscle was measured while cooling the sample stored at -80 ° C with liquid nitrogen. After the weight measurement, the sample was thawed for 24 hours in a low-temperature room in a tube. The moisture exuded from the thawed muscle tissue was wiped with a paper towel and weighed, and the shear strength was measured with a rheometer (RE-3305S, manufactured by YAMADEN, shear rate 1 mm / sec).
  • ⁇ Experiment method> Experimental animals Four-week-old Wistar male rats were used as animals. The experiment was conducted after one week of pre-breeding. Rats were divided into fish oil group or lard group (6 animals per group) and allowed to eat freely for 4 weeks. Water was ad libitum. The animals were housed in individual plastic plastic cages in an air-conditioned animal room with a room temperature of 22 ⁇ 2 ° C, 50 ⁇ 10% humidity, and illumination for 12 hours. 2.
  • soybean oil contained in AIN-93G is fish oil (fish oil produced by Nippon Suisan Co., Ltd., high content of EPA / DHA, 40% or more of total fatty acids) or lard ( AIN-93G modified feed of powder replaced with Oriental Yeast Co., Ltd.) was prepared.
  • a powder feed obtained by removing soybean oil from AIN-93G was obtained from Oriental Yeast Co., Ltd.
  • Table 3 shows the fatty acid composition of the fish oil used
  • Table 4 shows the component composition of the feed used. The same lard as in Table 1 was used.
  • the feeding period was 4 weeks. After the breeding period, the animals were dissected and the weight of each organ was measured and the shear strength of skeletal muscle was measured. In addition, during pre-breeding, thigh muscle hardness was measured at 2 weeks after the start of test feed intake and 4 weeks after the start of intake.
  • Shear strength measurement At the time of dissection, the anterior tibialis and rectus femoris were collected, the muscles were cut with a cutter with both ends fixed with two pins, and the shear strength at that time was measured with a rheometer (RE -3305S, manufactured by YAMADEN, shear rate 1 mm / sec).
  • Fig. 3 shows the correlation between rectus femoris shear strength and perirenal fat weight 4 weeks after the start of test feed intake.
  • composition of the present invention has the effect of softening muscles by oral administration, it can be used in the sports field and geriatric field by taking it as a pharmaceutical or a supplement.

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Abstract

Provided is a composition that improves the flexibility of muscles by oral intake. The composition, which makes muscles flexible by oral intake, comprises an n-3 highly unsaturated fatty acid as an active ingredient. The n-3 highly unsaturated fatty acid is preferably in the form of a free fatty acid, a lower alcohol ester or a glyceride or phospholipid containing the n-3 highly unsaturated fatty acid as a constituting fatty acid. Particular examples thereof include eicosapentaenoic acid, docosapentaenoic acid, docosahexaenoic acid, purified fish oil, purified krill oil and purified microbial oil.

Description

筋肉を柔軟にするための組成物Composition for softening muscles
　本発明は、筋肉を柔軟にするための組成物に関する。特に、運動機能に影響を及ぼすとされる骨格筋の柔軟性を高めるために経口摂取するための組成物である。 The present invention relates to a composition for softening muscles. In particular, it is a composition for ingestion in order to increase the flexibility of skeletal muscle, which is said to affect motor function.
　筋柔軟性は、健康づくりや傷害予防の重要な要因であり、筋柔軟性の維持・改善によりスポーツ傷害を予防・軽減することができると考えられている（非特許文献１）。また、筋柔軟性とスポーツパフォーマンスにも関連性があり、筋柔軟性をストレッチングで改善することで走行経済性が高まることがわかっている（非特許文献１）。従って、筋柔軟性を改善することは、傷害の予防・改善ならびに運動パフォーマンスの向上につながると考えられる（非特許文献１）。 Muscle flexibility is an important factor for health promotion and injury prevention, and it is considered that sports injury can be prevented and reduced by maintaining and improving muscle flexibility (Non-Patent Document 1). In addition, muscle flexibility and sports performance are also related, and it is known that running economy is improved by improving muscle flexibility by stretching (Non-patent Document 1). Therefore, it is considered that improving muscle flexibility leads to prevention / improvement of injury and improvement of exercise performance (Non-Patent Document 1).
　魚油にはエイコサペンタエン酸（以下、EPAとも記す）やドコサヘキサエン酸（以下、DHAとも記す）などの高度不飽和脂肪酸（以下、PUFAとも記す）が豊富に含まれているが、それらをヒトが摂取することで血液粘度が低下し、心筋梗塞や脳梗塞等の循環器系疾患の発症が予防できると考えられている（非特許文献２）。非特許文献３には、EPAが筋肉の蛋白合成を刺激する可能性について報告されている。 Fish oil is rich in polyunsaturated fatty acids (hereinafter also referred to as PUFA) such as eicosapentaenoic acid (hereinafter also referred to as EPA) and docosahexaenoic acid (hereinafter also referred to as DHA). By doing so, it is considered that blood viscosity is reduced and the onset of cardiovascular diseases such as myocardial infarction and cerebral infarction can be prevented (Non-patent Document 2). Non-Patent Document 3 reports the possibility that EPA stimulates muscle protein synthesis.
　本発明は、スポーツ分野や老年医学の分野で注目される、筋肉の柔軟性に着目し、運動やマッサージなどにより柔軟性を高める方法をサポートするものとして、経口摂取することにより、筋肉の柔軟性を高める組成物を提供することを課題とする。 The present invention focuses on muscle flexibility, which is attracting attention in the field of sports and geriatrics, and supports muscle flexibility by exercising and massage, etc. It is an object of the present invention to provide a composition that enhances the viscosity.
　PUFAの心血管系に対する機能のメカニズムの一つとして、粘度の低いPUFAが赤血球膜に取込まれることで赤血球が柔軟になり、狭い血管でも血流がスムーズになることが知られている。このPUFAの作用に着想を得て、PUFAを摂取することでPUFAが骨格筋に取込まれ、骨格筋の柔軟性が改善するとの仮説を立てた。魚油を数週間食べさせたラットの骨格筋の柔軟性を筋硬度計及びレオメーターを用いて計測したところ、比較対照であるラードを食べさせたラットよりも骨格筋の柔軟性が高くなることを確認し、本発明を完成させた。 As one of the functional mechanisms of PUFA for the cardiovascular system, it is known that PUFA having a low viscosity is taken into the erythrocyte membrane, so that red blood cells become flexible and blood flow is smooth even in narrow blood vessels. Inspired by the action of this PUFA, we hypothesized that ingestion of PUFA would bring PUFA into skeletal muscle and improve skeletal muscle flexibility. The flexibility of the skeletal muscle of rats fed with fish oil for several weeks was measured using a muscle hardness meter and rheometer. Confirmed and completed the present invention.
　本発明は、下記（１）～（８）の経口摂取により筋肉を柔軟化させるための組成物を要旨とする。
（１）ｎ－３系高度不飽和脂肪酸を有効成分として含有する経口摂取により筋肉を柔軟化させるための組成物。
（２）ｎ－３系高度不飽和脂肪酸が、遊離脂肪酸、低級アルコールエステル、又はｎ－３系高度不飽和脂肪酸を構成脂肪酸として含有するグリセリド又はリン脂質である（１）の組成物。
（３）ｎ－３系高度不飽和脂肪酸がエイコサペンタエン酸、ドコサペンタエン酸、又はドコサヘキサエン酸である（１）又は（２）の組成物。
（４）ｎ－３系高度不飽和脂肪酸を精製魚油、精製オキアミ油、精製微生物油として含有する（１）の組成物。
（５）筋肉が骨格筋である（１）ないし（４）いずれかの組成物。
（６）１日量として、ｎ－３系高度不飽和脂肪酸を合計で５～１５０ｍｇ／ｋｇ摂取させるための（１）ないし（５）いずれかの組成物。 The gist of the present invention is the following composition (1) to (8) for softening muscles by oral ingestion.
(1) A composition for softening muscles by ingestion containing an n-3 polyunsaturated fatty acid as an active ingredient.
(2) The composition according to (1), wherein the n-3 highly unsaturated fatty acid is a free fatty acid, a lower alcohol ester, or a glyceride or phospholipid containing n-3 highly unsaturated fatty acid as a constituent fatty acid.
(3) The composition according to (1) or (2), wherein the n-3 highly unsaturated fatty acid is eicosapentaenoic acid, docosapentaenoic acid, or docosahexaenoic acid.
(4) The composition according to (1), containing n-3 polyunsaturated fatty acids as refined fish oil, refined krill oil, and refined microbial oil.
(5) The composition according to any one of (1) to (4), wherein the muscle is skeletal muscle.
(6) The composition according to any one of (1) to (5) for ingesting 5 to 150 mg / kg of n-3 polyunsaturated fatty acids as a daily dose.
　本発明の組成物を経口摂取することにより、筋肉、特に骨格筋の柔軟性を高めることができる。 ¡By taking the composition of the present invention orally, the flexibility of muscles, particularly skeletal muscles, can be increased.
実施例１で採取したラットの前脛骨筋の剪断力価を示す図である。It is a figure which shows the shear strength of the anterior tibial muscle of the rat extract | collected in Example 1. FIG. 実施例２におけるラット大腿部の筋硬度変化率を示す図である。It is a figure which shows the muscle hardness change rate of the rat thigh in Example 2. FIG. 実施例２で採取したラットにおける大腿直筋の剪断力価と腎周囲脂肪重量との相関関係を示す図である。It is a figure which shows the correlation with the shear strength of a rectus femoris muscle in the rat extract | collected in Example 2, and perinephric fat weight.
　本発明において、PUFAとは、炭素数１８以上、二重結合数３以上の脂肪酸であり、好ましくは炭素数２０以上、二重結合数３以上の脂肪酸、さらに好ましくは炭素数２０以上、二重結合数４以上の脂肪酸である。n-3系PUFAとしては、α－リノレン酸、エイコサペンタエン酸、ドコサペンタエン酸、又はドコサヘキサエン酸が例示される。
　本発明で用いるPUFAは、遊離脂肪酸、低級アルコールエステル、又はｎ－３系高度不飽和脂肪酸を構成脂肪酸として含有するグリセリド又はリン脂質などの形態で用いることができる。好ましくは、グリセリンとのエステル、すなわち、トリグリセリド、ジグリセリド、モノグリセリド、あるいはリン脂質、すなわち、ホスファチジルコリン、ホスファチジルセリン、ホスファチジルエタノールアミン、ホスファチジルイノシトール、ホスファチジルグリセロール、カルジオリピン、ホスファチジン酸（いずれにもリゾ体を含む）、あるいは、低級アルコール（炭素数１～３）とのエステル、すなわち、メチルエステル、エチルエステルなどである。EPA、DHAなどn-3系の脂肪酸を主成分とし、実質的にn-6系の脂肪酸を含まないものが好ましい。具体的にはn-6系の脂肪酸の合計が１０重量％以下、好ましくは５重量％以下が好ましい。
　具体的には、トリグリセリドとしては精製魚油、精製微生物油、あるいはそれらに含まれるn-3系PUFAの濃度を高めた濃縮油（酵素反応、化学反応により精製魚油のn-3系PUFAの濃度を濃縮したもの）などが使用できる。また、リン脂質としては、精製オキアミ油、精製微生物油などを使用できる。EPA、DHAを含むn-3系PUFA源としては、魚介類抽出物、動物抽出物、卵黄抽出物、植物抽出物、菌類抽出物等、好ましくはオキアミ油、魚油、イカ抽出物、カツオ卵巣抽出物、遺伝子組み換えをした植物の抽出物等、ラビリンチュラ類の抽出物等が挙げられる。リン脂質を多く含む素材として、オキアミ油、イカ抽出物、が挙げられる。当該技術分野において一般的に知られている濃縮、抽出、及び／又は精製、配合等の技術を用いることによって、これらの素材中のリン脂質濃度や純度を任意に調節することができる。 In the present invention, PUFA is a fatty acid having 18 or more carbon atoms and 3 or more double bonds, preferably a fatty acid having 20 or more carbon atoms and 3 or more double bonds, more preferably 20 or more carbon atoms and a double bond. It is a fatty acid having 4 or more bonds. Examples of the n-3 PUFA include α-linolenic acid, eicosapentaenoic acid, docosapentaenoic acid, and docosahexaenoic acid.
The PUFA used in the present invention can be used in the form of glycerides or phospholipids containing free fatty acids, lower alcohol esters, or n-3 polyunsaturated fatty acids as constituent fatty acids. Preferably, esters with glycerin, i.e. triglycerides, diglycerides, monoglycerides, or phospholipids, i.e. Or an ester with a lower alcohol (having 1 to 3 carbon atoms), that is, a methyl ester, an ethyl ester, or the like. Preference is given to those containing n-3 fatty acids as the main component, such as EPA and DHA, and substantially free of n-6 fatty acids. Specifically, the total of n-6 fatty acids is 10% by weight or less, preferably 5% by weight or less.
Specifically, as triglycerides, refined fish oil, refined microbial oil, or concentrated oil with a higher concentration of n-3 PUFA contained in them (enzyme reaction, chemical reaction to increase the concentration of n-3 PUFA in refined fish oil) Concentrated) can be used. Moreover, refined krill oil, refined microbial oil, etc. can be used as a phospholipid. N-3 PUFA sources containing EPA and DHA include fish and shellfish extracts, animal extracts, egg yolk extracts, plant extracts, fungal extracts, etc., preferably krill oil, fish oil, squid extract, skipjack ovary extract Products, extracts of genetically modified plants, extracts of Labyrinthula, and the like. Examples of the material containing a large amount of phospholipid include krill oil and squid extract. The phospholipid concentration and purity in these materials can be arbitrarily adjusted by using techniques generally known in the art such as concentration, extraction, and / or purification, and blending.
　EPA及び／又はDHAは、高脂血症やアレルギー症状などに有効であることが知られており、すでに医薬品や各種サプリメントが多く市販されている。本発明の目的にはそれらと同様のカプセル製剤を用いることができる。具体的には、精製魚油に抗酸化剤を添加した油脂をゼラチンカプセルに加工したものが多く利用されている。製剤の剤形としては経口投与可能な剤形であれば特に限定されず、錠剤、カプセル剤（ハードカプセル、ソフトカプセル）、マイクロカプセル剤、顆粒剤、細粒剤、散剤、経口用液体製剤（エマルジョン、懸濁液など）、シロップ剤、ゼリー剤などの形にすることができる。例えば、カプセル、特に、ソフトカプセルやマイクロカプセルに封入した剤形とすることが好ましい。各製剤は、当該技術分野において公知の方法により、通常使用される賦形剤、希釈剤、結合剤、付湿剤、崩壊剤、崩壊抑制剤、吸収促進剤、滑沢剤、溶解補助剤、緩衝剤、乳化剤、懸濁剤などを使用して調製される。また添加剤としては、剤形に応じて通常使用される安定化剤、保存剤、緩衝剤、等張化剤、キレート剤、ｐＨ調整剤、界面活性剤、着色剤、香料、風味剤、甘味剤などを使用することができる。製剤中に含まれるn-3系PUFA量は、n-3系PUFAの合計量として、例えば２０重量％以上、好ましくは４０重量％以上、あるいは５０重量％以上、より好ましくは８０重量％以上である。 EPA and / or DHA are known to be effective for hyperlipidemia and allergic symptoms, and many pharmaceuticals and various supplements are already on the market. For the purposes of the present invention, capsule preparations similar to these can be used. Specifically, a product obtained by processing a fat and oil obtained by adding an antioxidant to purified fish oil into a gelatin capsule is often used. The dosage form of the preparation is not particularly limited as long as it is an orally administrable dosage form, and is a tablet, capsule (hard capsule, soft capsule), microcapsule, granule, fine granule, powder, oral liquid preparation (emulsion, Suspensions, etc.), syrups, jellies and the like. For example, a dosage form enclosed in a capsule, particularly a soft capsule or a microcapsule is preferable. Each formulation is prepared by methods known in the art, and usually used excipients, diluents, binders, wetting agents, disintegrants, disintegration inhibitors, absorption enhancers, lubricants, solubilizers, Prepared using buffers, emulsifiers, suspending agents and the like. Additives include stabilizers, preservatives, buffers, isotonic agents, chelating agents, pH adjusters, surfactants, colorants, flavoring agents, flavoring agents, sweetening agents that are usually used depending on the dosage form. An agent or the like can be used. The amount of n-3 PUFA contained in the preparation is, for example, 20% by weight or more, preferably 40% by weight or more, or 50% by weight or more, more preferably 80% by weight or more as the total amount of n-3 PUFA. is there.
　精製魚油としては、EPA及びDHAを含有する魚油であれば、どの魚類の魚油であってもかまわない。EPAが多く含まれる魚油としてはイワシ油、タラ肝油など、DHAが多く含まれる魚油としてはマグロ油、カツオ油などが例示される。
　PUFAの濃縮油とは、位置選択性、脂肪酸選択性のあるリパーゼなどを用いて、精製魚油のトリグリセリド中のEPA/DHAの濃度を高めた油脂である。 As the refined fish oil, any fish oil may be used as long as it contains EPA and DHA. Examples of fish oils rich in EPA include sardine oil and cod liver oil, and examples of fish oils rich in DHA include tuna oil and bonito oil.
The concentrated oil of PUFA is a fat and oil in which the concentration of EPA / DHA in the triglyceride of refined fish oil is increased by using a lipase having regioselectivity and fatty acid selectivity.
　本発明の組成物は、EPA、DHAなどのn-3系PUFAを合計で1日あたり５～１５０ｍｇ／ｋｇ経口摂取するのが好ましい。さらに好ましくは１５～１５０ｍｇ／ｋｇ、特に好ましくは１５～７５ｍｇ／ｋｇである。一度に摂取しても分割して摂取してもよい。継続的に摂取することが好ましく、少なくとも２週間連続して投与することにより効果が認められる。
　n-3系PUFA は食品由来成分であり、また、すでに他の用途で長い使用経験がある成分なので、安全性の点でも安心して摂取できる。 In the composition of the present invention, n-3 PUFAs such as EPA and DHA are preferably taken orally in a total amount of 5 to 150 mg / kg per day. More preferably, it is 15 to 150 mg / kg, and particularly preferably 15 to 75 mg / kg. It may be taken at once or divided. It is preferable to ingest continuously, and the effect is recognized by continuous administration for at least 2 weeks.
n-3 PUFA is a food-derived ingredient, and since it has already been used for a long time in other applications, it can be safely consumed.
　本発明の組成物は、有効成分の安全性が極めて高いことから、使用対象は特に限定されず、若年者から高齢者まで、筋肉の柔軟化を望む対象者に使用することができる。特にスポーツを行う者、高齢により筋肉の硬化が気になる者などに用いるのが好ましい。また、使用対象として哺乳類の動物にも適用できる。特に馬や犬等の競技用動物に用いるのが好ましい。
　本発明の組成物は、有効成分の安全性が極めて高いことから、食品として、あるいは、食品の成分と共に摂取することができる。
　以下に本発明の実施例を記載するが、本発明はこれらに何ら限定されるものではない。 In the composition of the present invention, since the active ingredient is extremely safe, the use target is not particularly limited, and it can be used for subjects who want to soften muscles, from young people to elderly people. It is particularly preferable for those who perform sports or those who are concerned about muscle hardening due to age. It can also be applied to mammalian animals as objects of use. It is particularly preferable to use it for competition animals such as horses and dogs.
The composition of the present invention can be ingested as a food or together with a food ingredient because the active ingredient is extremely safe.
Examples of the present invention will be described below, but the present invention is not limited thereto.
＜実験方法＞
1. 実験動物
　動物は7 週齢Wistar 系雄ラットを用いた。実験は1 週間の予備飼育後に行った。ラットをEPA・DHA高含有魚油添加飼料摂取群（魚油群）とラード添加飼料摂取群（ラード群）の2群に分けた（1群6匹）。飼料はpair-feeding を行い、摂食量を揃えた。水は自由摂取させた。動物はステンレス製の個別ケージで、室温22±2℃、50±10%湿度、12 時間照明の空調動物室内にて飼育した。
2. 飼料と給餌期間
　魚油（日本化学飼料株式会社製）、ラード（オリエンタル酵母工業株式会社製）がそれぞれ15%含まれる、ペレット状に固形化したAIN-93G 改変飼料をオリエンタル酵母工業（株）より入手した。用いた魚油及びラードの脂肪酸組成を表１に、AIN-93G 改変飼料の組成を表２に示した。給餌期間は4 週間とし、飼育期間終了後、前脛骨筋（TA）を採取し、分析まで－80℃で凍結保存した。 <Experiment method>
1. Experimental animals 7-week-old Wistar male rats were used as animals. The experiment was conducted after one week of pre-breeding. Rats were divided into 2 groups (6 groups), EPA / DHA-rich fish oil-added feed intake group (fish oil group) and lard-added feed intake group (Lard group). The feed was pair-feeded and the food intake was adjusted. Water was ad libitum. The animals were housed in individual cages made of stainless steel in a conditioned animal room with a room temperature of 22 ± 2 ° C., 50 ± 10% humidity, and illumination for 12 hours.
2. Feed and feeding period AIN-93G modified feed solidified in pellet form containing 15% each of fish oil (manufactured by Nippon Chemical Feed Co., Ltd.) and lard (manufactured by Oriental Yeast Co., Ltd.) Obtained from. Table 1 shows the fatty acid composition of the fish oil and lard used, and Table 2 shows the composition of the AIN-93G modified feed. The feeding period was 4 weeks. After the breeding period, the anterior tibial muscle (TA) was collected and stored frozen at −80 ° C. until analysis.
Figure JPOXMLDOC01-appb-T000001
Figure JPOXMLDOC01-appb-T000001
Figure JPOXMLDOC01-appb-T000002
Figure JPOXMLDOC01-appb-T000002
3. 剪断力価測定
　－80℃保存下のサンプルを液体窒素で冷やしながら、前脛骨筋の重量を測定した。重量測定後、チューブに入れた状態で低温室にて24時間解凍した。解凍した筋組織からしみ出た水分をペーパータオルで拭って重量測定し、剪断力価をレオメーター（RE-3305S、YAMADEN社製、剪断速度 1 mm/秒）により測定した。 3. Shear titration measurement The weight of the anterior tibial muscle was measured while cooling the sample stored at -80 ° C with liquid nitrogen. After the weight measurement, the sample was thawed for 24 hours in a low-temperature room in a tube. The moisture exuded from the thawed muscle tissue was wiped with a paper towel and weighed, and the shear strength was measured with a rheometer (RE-3305S, manufactured by YAMADEN, shear rate 1 mm / sec).
＜結果＞
　４週間の飼料の総摂取量は魚油群708.9±31.4 g、ラード群695.1±27.6 gであった。魚油群の一日あたりのEPAおよびDHAの摂取量は、それぞれ約765.8mgと約307.8mgであった。また、魚油群及びラード群の前脛骨筋の重量はそれぞれ1.42±0.05 g及び1.42±0.02 gであった。
　魚油群及びラード群の剪断力価を測定したところ、魚油群の剪断力価が、ラード群の剪断力価と比較して小さい傾向が観察された（図１）。魚油の摂取は骨格筋の柔軟性を高めることが示された。 <Result>
The total intake of feed for 4 weeks was 708.9 ± 31.4 g in the fish oil group and 695.1 ± 27.6 g in the lard group. The daily intake of EPA and DHA in the fish oil group was about 765.8 mg and about 307.8 mg, respectively. The weights of the anterior tibial muscles in the fish oil group and lard group were 1.42 ± 0.05 g and 1.42 ± 0.02 g, respectively.
When the shear strength of the fish oil group and lard group was measured, a tendency was observed that the shear strength of the fish oil group was smaller than the shear strength of the lard group (FIG. 1). Fish oil intake has been shown to increase skeletal muscle flexibility.
＜実験方法＞
1. 実験動物
　動物は4 週齢Wistar 系雄ラットを用いた。実験は1 週間の予備飼育後に行った。ラットを魚油群又はラード群に分け（1群6匹）、4週間自由摂食させた。水は自由摂取させた。動物はプラスチック製の個別ケージで、室温22±2℃、50±10%湿度、12 時間照明の空調動物室内にて飼育した。
2. 飼料と給餌期間
　AIN-93G中に含まれる7%の大豆油を魚油（日本水産株式会社製のEPA・DHA高含有魚油、EPA・DHAを総脂肪酸中40%以上含有）、又はラード（オリエンタル酵母工業株式会社製）に置換した粉末のAIN-93G 改変飼料を作成した。AIN-93Gから大豆油を抜いた粉末飼料はオリエンタル酵母工業（株）より入手した。用いた魚油の脂肪酸組成を表３に、用いた飼料の成分組成を表４に示した。ラードは表１と同じものを用いた。給餌期間は4 週間とし、飼育期間終了後、解剖し、各臓器重量の測定、並びに骨格筋の剪断力価測定を行った。また、予備飼育中、試験飼料摂取開始後2週、摂取開始後4週の時点で大腿部の筋硬度を測定した。 <Experiment method>
1. Experimental animals Four-week-old Wistar male rats were used as animals. The experiment was conducted after one week of pre-breeding. Rats were divided into fish oil group or lard group (6 animals per group) and allowed to eat freely for 4 weeks. Water was ad libitum. The animals were housed in individual plastic plastic cages in an air-conditioned animal room with a room temperature of 22 ± 2 ° C, 50 ± 10% humidity, and illumination for 12 hours.
2. Feed and Feeding Period 7% soybean oil contained in AIN-93G is fish oil (fish oil produced by Nippon Suisan Co., Ltd., high content of EPA / DHA, 40% or more of total fatty acids) or lard ( AIN-93G modified feed of powder replaced with Oriental Yeast Co., Ltd.) was prepared. A powder feed obtained by removing soybean oil from AIN-93G was obtained from Oriental Yeast Co., Ltd. Table 3 shows the fatty acid composition of the fish oil used, and Table 4 shows the component composition of the feed used. The same lard as in Table 1 was used. The feeding period was 4 weeks. After the breeding period, the animals were dissected and the weight of each organ was measured and the shear strength of skeletal muscle was measured. In addition, during pre-breeding, thigh muscle hardness was measured at 2 weeks after the start of test feed intake and 4 weeks after the start of intake.
Figure JPOXMLDOC01-appb-T000003
Figure JPOXMLDOC01-appb-T000003
Figure JPOXMLDOC01-appb-T000004
Figure JPOXMLDOC01-appb-T000004
3. 筋硬度測定
　ラットを保定し、右後足の大腿部の筋硬度を筋硬度計（NEUTONE TDN-N1、TRY-ALL社製）を用いて測定した。測定は1匹につき連続して3回行い、3回の平均値を測定値とした。予備飼育中の筋硬度の値を基準にして100%とし、その後の筋硬度変化を変化率で表した。
4. 剪断力価測定
　解剖時に前脛骨筋、大腿直筋を採取し、2本のピンで筋肉の両端を固定した状態でカッターにより筋肉を切断し、その際の剪断力価をレオメーター（RE-3305S、YAMADEN社製、剪断速度 1 mm/秒）で測定した。 3. Measurement of muscle hardness Rats were held and the muscle hardness of the thigh of the right hind paw was measured using a muscle hardness meter (NEUTONE TDN-N1, manufactured by TRY-ALL). The measurement was carried out 3 times in succession for each animal, and the average value of 3 times was taken as the measured value. Based on the value of muscle hardness during pre-breeding, it was set to 100%, and the subsequent change in muscle hardness was expressed as the rate of change.
4. Shear strength measurement At the time of dissection, the anterior tibialis and rectus femoris were collected, the muscles were cut with a cutter with both ends fixed with two pins, and the shear strength at that time was measured with a rheometer (RE -3305S, manufactured by YAMADEN, shear rate 1 mm / sec).
＜結果＞
　1匹当りの4週間の総摂取量は魚油群が約544gであり、ラード群は約585gであった。魚油群の一日あたりのEPAおよびDHAの摂取量は、それぞれ約345.0mgと約153.5mgであった。 <Result>
The total intake for 4 weeks per animal was about 544 g in the fish oil group and about 585 g in the lard group. The daily intake of EPA and DHA in the fish oil group was about 345.0 mg and about 153.5 mg, respectively.
　筋硬度変化率の結果を図２に示した。魚油群では試験飼料摂取開始後2週及び4週において大腿部の筋硬度が経時的に柔らかくなっていた。一方でラード群では摂取後2週の時点では摂取前と変化が見られず、摂取後4週で10%程度柔らかくなっていた。2群間で比較すると、摂取開始後2週の時点で魚油群の筋硬度が有意に低くなった。これらの結果より、ラード食と比較して魚油食では筋肉の柔軟性が向上することが示された。
　試験飼料摂取後4週時点の前脛骨筋及び大腿直筋の剪断力価を表５に示した。前脛骨筋及び大腿直筋において魚油群の剪断力価はラード群と比較して低値を示した。これらの結果も、ラード食と比較して魚油食では筋肉の柔軟性が向上することを示すものであった。 The results of the muscle hardness change rate are shown in FIG. In the fish oil group, the muscle hardness of the thigh became softer over time at 2 and 4 weeks after the start of test feed intake. On the other hand, in the lard group, there was no change from before intake at 2 weeks after ingestion, and it became softer by about 10% at 4 weeks after ingestion. When compared between the two groups, the muscle hardness of the fish oil group was significantly lower at 2 weeks after the start of intake. From these results, it was shown that the fish oil diet improves muscle flexibility compared to the lard diet.
Table 5 shows the shear titers of the anterior tibial muscle and rectus femoris at 4 weeks after the intake of the test feed. In the tibialis anterior and rectus femoris, the fish oil group showed lower shear strength than the lard group. These results also showed that the fish oil diet improved muscle flexibility compared to the lard diet.
Figure JPOXMLDOC01-appb-T000005
Figure JPOXMLDOC01-appb-T000005
　試験飼料摂取開始後4週時点の大腿直筋の剪断力価と腎周囲脂肪重量の相関関係を図３に示した。ラード群においては両者に相関関係は見られなかったが、魚油群においては有意な正の相関関係が見られた。これらの結果は、魚油摂取により腎周囲脂肪重量が減った個体ほど大腿直筋が柔らかくなったことを示しており、魚油に対する感受性が高い個体ほど筋肉の柔軟性も高まることを示唆している。この結果もラード食と比較して魚油食では筋肉柔軟性が高まることを支持する結果であった。以上の結果より、PUFAの摂取は骨格筋の柔軟性を高めると考えられた。 Fig. 3 shows the correlation between rectus femoris shear strength and perirenal fat weight 4 weeks after the start of test feed intake. There was no correlation between the two in the lard group, but there was a significant positive correlation in the fish oil group. These results indicate that the rectus femoris muscle became softer as the weight of the perirenal fat decreased with fish oil intake, suggesting that the individual with higher sensitivity to fish oil increases muscle flexibility. This result also supported the increased muscle flexibility in the fish oil diet compared to the lard diet. Based on the above results, it was considered that the intake of PUFA increases the flexibility of skeletal muscle.
　スポーツ分野や老年医学の分野では骨格筋の硬直化は好ましいものではないと考えられており、トレーニングやマッサージにより柔軟にすることが行われている。本発明の組成物は、経口投与により、筋肉を柔軟にする作用を有するので、医薬品やサプリメントとして摂取することにより、スポーツ分野や老年医学分野で利用できる。
  In the field of sports and geriatric medicine, skeletal muscle stiffness is considered unfavorable and has been made flexible by training and massage. Since the composition of the present invention has the effect of softening muscles by oral administration, it can be used in the sports field and geriatric field by taking it as a pharmaceutical or a supplement.

Claims (6)

  1. 　ｎ－３系高度不飽和脂肪酸を有効成分として含有する経口摂取により筋肉を柔軟化させるための組成物。 A composition for softening muscles by ingestion containing an n-3 polyunsaturated fatty acid as an active ingredient.
  2. 　ｎ－３系高度不飽和脂肪酸が、遊離脂肪酸、低級アルコールエステル、又はｎ－３系高度不飽和脂肪酸を構成脂肪酸として含有するグリセリド又はリン脂質である請求項１の組成物。 2. The composition according to claim 1, wherein the n-3 highly unsaturated fatty acid is a free fatty acid, a lower alcohol ester, or a glyceride or phospholipid containing n-3 highly unsaturated fatty acid as a constituent fatty acid.
  3. 　ｎ－３系高度不飽和脂肪酸がエイコサペンタエン酸、ドコサペンタエン酸、又はドコサヘキサエン酸である請求項１又は２の組成物。 3. The composition according to claim 1 or 2, wherein the n-3 highly unsaturated fatty acid is eicosapentaenoic acid, docosapentaenoic acid, or docosahexaenoic acid.
  4. 　ｎ－３系高度不飽和脂肪酸を精製魚油、精製オキアミ油、精製微生物油として含有する請求項１の組成物。 The composition according to claim 1, comprising n-3 polyunsaturated fatty acids as refined fish oil, refined krill oil, and refined microbial oil.
  5. 　筋肉が骨格筋である請求項１ないし４いずれかの組成物。 The composition according to any one of claims 1 to 4, wherein the muscle is skeletal muscle.
  6. 　１日量として、ｎ－３系高度不飽和脂肪酸を合計で５～１５０ｍｇ／ｋｇ摂取させるための請求項１ないし５いずれかの組成物。
     

          The composition according to any one of claims 1 to 5, which is used for daily intake of 5 to 150 mg / kg of n-3 polyunsaturated fatty acids.


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