JP6225199B2 - Motor function improver - Google Patents

Description

  The present invention relates to a motor function improving agent that exhibits a motor function improving effect.

  In general, exercise training and balanced nutrition are considered important for improving motor functions such as endurance and muscle strength. In recent years, exercise enthusiasts and athletes have attempted to use nutrition supplements such as supplements in addition to training to improve muscle strength more efficiently (Patent Document 1). However, there is a concern that training with excessive intake of some proteins and amino acids may cause adverse effects on renal function and the like (Non-patent Document 1).

  In addition to exercise enthusiasts and athletes, muscular strength and weakness due to dystrophy and muscle weakness due to aging and muscle non-use, due to insufficient dietary supply of nutrients due to excessive diet, It is regarded as a problem that various motor functions such as endurance decline, and further, fatigue due to the decline of motor function.

  Therefore, efficient motor function improvement technology is desired not only for exercise enthusiasts and athletes aiming at performance improvement but also for general people.

  From such a viewpoint, a component having a motor function improving action is searched, for example, endurance improving action by tea catechin (Patent Document 2), polymer fruit polyphenol (Patent Document 3) and phytic acid (Patent Document 4). It has been reported that muscle strength is improved.

  Furthermore, in recent years, it has been clarified that phospholipids may affect motor function. Inhibition of lactic acid accumulation during exercise by phosphatidylcholine (Non-patent Document 2) and endurance improvement effect by phosphatidylserine (Non-patent Document) 3) etc. have been reported.

  On the other hand, sphingomyelin is a substance having a structure in which phosphocholine is bound to a ceramide skeleton composed of a sphingoid base and a fatty acid, and is known to exist in the brain and nerve tissues in the body. In recent years, research on the physiological function has been promoted, such as an effect of promoting the maturation and development of the digestive tract (Patent Document 5), an effect of improving learning ability (Patent Document 6), an effect of promoting the secretion of sialomucin (Patent Document 7), etc. It is known to have the physiological function of

  However, the effects of sphingomyelin on motor functions such as muscular strength and endurance have not been known so far.

Japanese Patent Laid-Open No. 2002-065212 JP 2005-89384 A International Publication No. 2005/07496 Pamphlet JP 2009-107987 A JP 2000-250563 A JP 2007-246404 A JP 2007-112793 A

Anderson, JAMA, 223, 1973 Von Allwarden, Phospholipids, AOCS Press, 1995 Kingsley, Med Sci Sports Excel, 38, 2006

  The present invention relates to providing a material that is used in pharmaceuticals, quasi-drugs, foods, and feeds that have abundant dietary experience, are highly safe, and have excellent motor function improvement effects.

  As a result of searching for an effective component in improving the motor function, the present inventors have an effect of improving endurance, improving muscular strength, or suppressing muscular strength in sphingomyelin, and this can exert the effect. The present invention was completed by finding it useful as an active ingredient of pharmaceuticals, quasi drugs, foods and feeds.

That is, the present invention relates to the following (1) to (4).
(1) An endurance improver comprising sphingomyelin as an active ingredient.
(2) A muscle strength improver comprising sphingomyelin as an active ingredient.
(3) A muscular strength reduction inhibitor comprising sphingomyelin as an active ingredient.
(4) An anti-fatigue agent containing sphingomyelin as an active ingredient.

  The endurance improver, anti-fatigue agent, muscle strength improver, and muscle weakness inhibitor of the present invention are used in a wide range of ages including the elderly to improve endurance and anti-fatigue during daily activities including exercise and work. It is useful as a material for blending as an active ingredient in foods, pharmaceuticals, quasi drugs or feeds for improving muscle strength or suppressing muscle strength reduction.

The graph which shows transition of swimming endurance. Ex represents a standard food intake + exercise group, and SPM represents a 0.25% sphingomyelin food intake + exercise group. The graph which shows the muscular strength of the extracted soleus and long finger extensor. Cont is a standard diet intake group, Ex is a standard diet intake + exercise group, and SPM is a 0.25% sphingomyelin diet intake + exercise group. The graph which shows the muscular strength of the isolated soleus. Normal represents a standard diet intake + untreated (non-tail suspension) group, Cont represents a standard diet intake + tail suspension group, and SPM represents a 0.25% sphingomyelin diet intake + tail suspension group. # Indicates a significant difference with respect to the Normal group.

The sphingomyelin that can be used in the present invention is not particularly limited, and examples thereof include chemically synthesized and naturally derived ones.
For example, as a chemical synthesis method of sphingomyelin, 1) a method via phosphoramidite (Weis, Chem Phys Lip, 3, 1999), 2) a method via cyclic phosphate (Dong, Tetrahedron Lett, 5291, 1991) Or 3) A method of converting to sphingomyelin by introducing phosphocholine into the 1-position hydroxyl group of ceramide by a method via a cyclic phosphite (Byun, J Org Chem, 6495, 1994) is known.
Furthermore, milk fat globule membrane components obtained from milk are purified by techniques such as dialysis, ammonium sulfate fractionation, gel filtration, isoelectric precipitation, ion exchange chromatography, solvent fractionation (Sanchez-Juanes, Int Dairy J, 273). , 2009), high-purity sphingomyelin can be obtained.
Furthermore, a commercial item can also be used as sphingomyelin. Examples of such commercially available products include NOF Corporation “milk-derived sphingomyelin: NM-70” and “egg yolk-derived sphingomyelin: NM-10”.

As shown in the examples below, sphingomyelin significantly increased swimming time and significantly increased the strength of the soleus muscle in mice. Has anti-fatigue action that is backed by improvement. In addition, sphingomyelin significantly suppresses the decrease in muscle strength in a mouse whose muscle strength is reduced by non-muscle use (tail suspension) treatment, and thus has an effect of suppressing muscle strength reduction.
Therefore, sphingomyelin can be used as an endurance improver, muscle strength improver, anti-fatigue agent, muscle strength reduction inhibitor (hereinafter referred to as “motor function improver etc.”), and these agents are also produced. Can be used to At this time, in the motor function improving agent, etc., sphingomyelin alone or in addition to this, other phospholipids, carriers, stabilizers, etc., which are appropriately selected as necessary, should be added as described later. In this case, an acceptable material may be used. In addition, the said formulation can be manufactured by a conventional method according to the target object which should be mix | blended.

  And the motor function improving agent of the present invention is effective for human or veterinary drugs, quasi-drugs, foods, or feeds that exert the effects of improving endurance, improving muscular strength, anti-fatigue, and suppressing muscular weakness. It can be used as a component. Further, the motor function improving agent of the present invention is based on the concept of improving endurance, improving muscle strength, anti-fatigue, and suppressing muscle weakness in those who are underexercise, middle-aged and elderly, bed rest persons, or athletes and exercise enthusiasts. It can be applied to foods, functional foods, foods for the sick, and foods for specified health use.

  Examples of the dosage form when the motor function improving agent of the present invention is used as an active ingredient of pharmaceuticals and quasi-drugs include oral administration or injections such as tablets, capsules, granules, powders, and syrups. Parenteral administration such as an agent, an inhalation agent, a transdermal absorption agent, and an external preparation. In addition, in order to prepare such various dosage forms, the motor function improving agent of the present invention alone or other pharmaceutically acceptable excipients, binders, extenders, disintegrants. , Surfactants, lubricants, dispersants, buffers, preservatives, flavoring agents, fragrances, coating agents, carriers, diluents, medicinal components other than fat globule film components, and the like can be used in appropriate combinations. Of these dosage forms, the preferred form is oral administration, and when an oral liquid preparation is prepared, it can be produced by a conventional method by adding a flavoring agent, a buffering agent, a stabilizer and the like.

Examples of the case where the motor function improving agent of the present invention is used as an active ingredient of food include milk, processed milk, milk beverage, yogurt, soft drink, tea-based beverage, coffee beverage, fruit juice beverage, carbonated beverage, juice In addition to foods and drinks such as jelly, wafers, biscuits, bread, noodles, sausages and various foods such as nutritional foods, nutritional supplements in the same form (tablets, capsules, syrups, etc.) as the above-mentioned oral preparations Composition for use.
To prepare various forms of food, the motor function improving agent of the present invention alone, or other food materials, solvents, softeners, oils, emulsifiers, preservatives, fragrances, stabilizers, colorants , Antioxidants, moisturizers, thickeners, active ingredients other than fat globule film components, etc., in combination as appropriate, food for improving motor function, food for improving endurance, food for anti-fatigue, food for improving muscle strength, pet food, etc. It is possible to blend in.
In addition, the motor function improving agent of the present invention can be blended in the form of a nutritional composition such as an enteral nutrient in elderly people who are difficult to supplement with an appropriate amount of nutrition or in bed rest. is there.
The amount of sphingomyelin in a beverage containing the motor function improving agent of the present invention, for example, a milk beverage, a soft drink, or a tea-based beverage (in terms of dry matter) is usually 0.0001 to 1.0% by mass, and further 0. 0.001 to 0.5% by mass, particularly 0.01 to 0.2% by mass is preferable.

  In the case of foods and feeds other than beverages containing the motor function improving agent of the present invention, and pharmaceuticals such as oral solid preparations such as tablets, granules and capsules, oral liquid preparations such as oral liquids and syrups The sphingomyelin (in terms of dry matter) is usually 0.002 to 50% by mass, more preferably 0.02 to 25% by mass, and particularly preferably 0.2 to 10% by mass. In addition, sphingomyelin may be in a dissolved state or in a dispersed state, and the presence state thereof does not matter.

  The intake of the motor function improving agent and the like of the present invention varies depending on the dosage form and application, but sphingomyelin is preferably 0.1 to 1000 mg / 60 kg body weight per day for an adult, What is necessary is just to adjust the compounding quantity etc. to a medicine, food-drinks, and a feed so that it may become 250 mg / 60 kg body weight and also 5-100 mg / 60 kg body weight.

  As typical examples of the present invention, Production Examples, Test Examples, and Formulation Examples are shown below, but the present invention is not limited thereto.

Production Example 1: Preparation of sphingomyelin Take 201 g of milk phospholipid PC-500 (obtained from Fontera Japan, containing 8.8% sphingomyelin), add 4000 mL of acetone, and add a homomixer (TK auto homomixer, special under ice cooling) Dispersed by Kika Industries Co., Ltd. Thereafter, the acetone-soluble fraction (neutral lipid) was removed by centrifugation. To the obtained acetone insoluble fraction A, chloroform (800 mL), methanol (400 mL), and water (300 mL) were added, followed by liquid-liquid extraction, and a chloroform layer was collected. The chloroform layer was concentrated under reduced pressure to obtain 154 g of a chloroform fraction A.
To the obtained chloroform fraction A, 28.05 g of potassium hydroxide and 1000 mL of methanol were added, and the mixture was stirred under nitrogen at 37 ° C. for 15 hours for hydrolysis. After completion of the reaction, 2000 mL of chloroform and 750 mL of water were added, and the liquid was partitioned, and the chloroform layer was collected. The chloroform layer was concentrated under reduced pressure and then neutralized with 15 mL of acetic acid, and 400 mL of chloroform, 200 mL of methanol, and 150 mL of water were added, and liquid-liquid extraction was performed again. The chloroform layer was collected and concentrated under reduced pressure to obtain 90 g of chloroform fraction B.
To the obtained chloroform fraction B, 1200 mL of acetone was added, and the mixture was dispersed with a homomixer (TK auto homomixer, manufactured by Tokushu Kika Kogyo Co., Ltd.) under ice cooling. Thereafter, the acetone-soluble fraction (free fatty acid) was removed by centrifugation. The same operation was further repeated twice to obtain 44 g of acetone insoluble fraction B.
To the obtained acetone insoluble fraction B, 400 mL of hexane, 400 mL of methanol, 150 mL of water, and 50 mL of 28% ammonia water were added, liquid-liquid extraction was performed, and a hexane layer was collected. The hexane layer was further washed with 400 mL of methanol, 150 mL of water, and 50 mL of 28% aqueous ammonia. The obtained hexane layer was concentrated under reduced pressure to obtain 27 g of a hexane fraction.
Purification was performed by silica gel column chromatography using 22 g of the obtained hexane fraction. That is, after adsorbing the chloroform solution of the hexane fraction to 1 kg of silica gel (silica gel 60, manufactured by Merck), the contaminants were eluted with a chloroform / methanol mixed solvent, then eluted with 21 L of methanol, and purified sphingomyelin fraction 16 g (yield: 8%) was obtained.

Test Example 1: Endurance improvement and muscle strength improvement effect of sphingomyelin <Method> Sphingomyelin was extracted from milk phospholipid (PC-500, Fontera Japan) according to the method of Production Example 1 above.
After 1 week of pre-breeding, 9-week-old male BALB / c mice (Charles River Japan) were tested for body weight and swimming endurance. Depth: 90 × 45 × 45 cm, water depth: 38 cm, water temperature: 34 ° C. (Matsumoto, J Appl Physiol. 81: 1843-1849, 1996)). Ex group, SPM group) (8 animals in each group).
After grouping, the Cont and Ex mice had a control diet (10% lipid, 20% casein, 55.5% potato starch, 8.1% cellulose, 0.2% methionine, 2.2% vitamin (commodity Name: Vitamin-mixed AIN-76, Oriental Bioservice), 4% mineral (trade name: Mineral-mixed AIN-76, Oriental Bioservice), and mice in SPM group containing sphingomyelin according to Production Example 1 Diet (10% lipid, 20% casein, 55.25% potato starch, 8.1% cellulose, 0.2% methionine, 2.2% vitamin, 4% mineral, 0.25% sphingomyelin) for 13 weeks I was fed.
During the feeding period, in the Ex group and SPM group mice, the limit swimming time was measured once a week using a running water pool for mice. The limit swimming time was defined as the time from the start of swimming until the mouse was unable to rise to the surface for 7 seconds due to breathing at a flow rate of 7 L / min. During this time, swimming training (6 L / min, 30 min) was performed twice a week to acclimate the mouse to exercise.
After rearing for 13 weeks, it was subjected to dissection and the muscle strength of the extracted soleus and long extensor muscles was measured. The muscle strength of the isolated muscle was measured according to the method of Cannon et al. (Biomed Sci Instrument, 2005). Specifically, the soleus and long finger extensors were removed from the mouse, fixed to the transducer (WPI, FORT100) using a suture (# 5-0 silk), and in a Krebs solution at 37 ° C. (95% -O ₂ , 5% -CO ₂ aeration). Electrical stimulation of 40 Hz, 330 ms, and 10 V was applied from two platinum electrodes, and the signal obtained from the transducer was measured as muscle strength (g / mg muscle).

<Results> FIG. 1 shows the effect of sphingomyelin on swimming endurance. In sphingomyelin-administered mice, an improvement in endurance was observed early after the start of administration, and after 5 weeks from the start of administration, a significant improvement in endurance was observed relative to the Cont group. FIG. 2 shows the effect of sphingomyelin on muscle strength improvement. In sphingomyelin-treated mice, the muscle strength of the soleus and long extensor muscles showed significantly high values.
Endurance and muscle strength are typical motor functions for moving the body, and it is considered that resistance to physical fatigue is improved by improving motor functions. Therefore, in this test, sphingomyelin increased the exercise endurance and the muscle strength of the extracted muscle, and it was revealed that sphingomyelin is effective in improving endurance, improving muscle strength, and anti-fatigue.

Test Example 2: Sphingomyelin suppresses muscle weakness <Method> Male BALB / c mice (9 weeks old) are preliminarily raised for 1 week and divided into 3 groups (Normal group, Cont group, SPM group) based on body weight. (Each group n = 8).
After grouping, the Normal and Cont mice had a control diet (10% lipid, 20% casein, 55.5% potato starch, 8.1% cellulose, 0.2% methionine, 2.2% vitamins, 4% minerals), and mice in the SPM group also had a test meal (10% lipid, 20% casein, 55.25% potato starch, 8.1% cellulose, 0.2%) containing sphingomyelin according to Production Example 1. % Methionine, 2.2% vitamin, 4% mineral, 0.25% sphingomyelin) for 2 weeks.
After feeding the test diet for 2 weeks, the mice in the Cont group and the SPM group were subjected to tail suspension treatment, and the gravity load on the hindlimb muscle group (such as the soleus muscle) was eliminated. Muscles with reduced load exhibit disuse muscle atrophy and muscle mass and strength decrease. The normal group of mice received no tail suspension treatment.
Seven days after tail suspension treatment, the mice were subjected to dissection, and the muscle strength of the extracted soleus muscle was measured. The muscle strength of the extracted muscle was measured by the same method as in Test Example 1.

<Results> FIG. 3 shows the strength of the extracted soleus muscle. With tail suspension treatment, the muscle strength of the Cont group and the SPM group significantly decreased. On the other hand, the muscular strength of the SPM group was significantly higher than that of the Cont group. Therefore, from these results, the effect of sphingomyelin in suppressing muscle weakness was clarified.

Formulation Example Formulation 1 Jelly food for improving motor function Carrageenan and locust bean gum mixed gelling agent 0.65%, grapefruit 50% concentrated fruit juice 5.0%, citric acid 0.05%, vitamin C 0.05% , And sphingomyelin (NM-70, NOF Corporation) were mixed, adjusted to 100% by adding water, and dissolved at 65 ° C. Further, a small amount of grapefruit flavor was added and kept at 85 ° C. for 5 minutes, sterilized, and dispensed into a 100 mL container. Allow to stand for 8 hours and cool to 5 ° C while gradually cooling to obtain a jelly food containing sphingomyelin with good mouth meltability, fruit flavor and good texture when put into the mouth. It was.

Formulation Example 2 Tablet for improving motor function Formulation consisting of 180 mg of ascorbic acid, 50 mg of citric acid, 12 mg of aspartame, 24 mg of magnesium stearate, 120 mg of crystalline cellulose, 594 mg of lactose, and 120 mg of sphingomyelin (NM-10, manufactured by NOF Corporation) 2200 mg), a tablet was produced according to the Japanese Pharmacopoeia (general general rule “tablet”) to obtain a tablet containing sphingomyelin.

Formulation Example 3 Vitamin oral solution for improving motor function Taurine 800 mg, sucrose 2000 mg, caramel 50 mg, sodium benzoate 30 mg, vitamin B1 nitrate 5 mg, vitamin B2 20 mg, vitamin B6 20 mg, vitamin C 2000 mg, vitamin E 100 mg, vitamin D3 2000 IU, Nicotinamide 20 mg, purified sphingomyelin (Production Example 1) 50 mg, leucine 200 mg, isoleucine 100 mg, and valine 100 mg are dissolved in an appropriate amount of purified water, adjusted to pH 3 with an aqueous phosphoric acid solution, and further purified water is added. The total volume was 50 mL. This was sterilized at 80 ° C. for 30 minutes to obtain a drink for improving motor function containing sphingomyelin and amino acids.

Formulation Example 4 Milk Beverage for Improvement of Motor Function Milk Casein 3.4g, Isolated Soy Protein 1.67g, Dextrin 14.86g, Sucrose 1.3g, Soybean Oil 1.75g, Perilla Oil 0.18g, Soybean Phospholipid 0 .14 g, glycerin fatty acid ester 0.07 g, minerals 0.60 g, vitamins 0.06 g, purified sphingomyelin (Production Example 1) 100 mg, purified water is added and sterilized by retort according to a conventional method, and contains sphingomyelin A motor function improving beverage (100 mL) was obtained.

Claims (2)

  A muscle strength improver that is taken during exercise, containing sphingomyelin as an active ingredient.   A food for improving muscular strength that is taken during exercise, containing sphingomyelin as an active ingredient.

Images