JP2020176103A - Composition for preventing or recovering myofunction reduction, composition for preventing or recovering muscle mass reduction and composition for preventing or recovering muscle quality reduction - Google Patents

Abstract

To provide a novel composition for preventing or recovering myofunction reduction, capable of preventing or recovering myofunction reduction; a novel composition for preventing or recovering muscle mass reduction, capable of preventing or recovering muscle mass reduction; and a novel composition for preventing or recovering muscle quality reduction, capable of preventing or recovering muscle quality reduction.SOLUTION: A composition for preventing or recovering myofunction reduction has a peptide including an amino acid sequence of His-Lys at a C terminal or a salt thereof as an active ingredient.SELECTED DRAWING: None

Description

The present invention relates to a composition for suppressing or recovering muscle function decline. The present invention also relates to a composition for suppressing or recovering muscle mass loss, and a composition for suppressing or recovering muscle loss.

In a country like Japan where the proportion of elderly people is high, extending healthy life expectancy and improving the quality of life (QOL) are issues. One of the factors for shortening healthy life expectancy is a decrease in muscle mass (muscle mass) and muscle quality (muscle quality) with aging. Decreased muscle mass and quality cause a decrease in muscle function, leading to an increased risk of falls, injuries, and long-term bed rest. When the amount of activity of the elderly decreases due to injury, illness, etc., it causes a vicious cycle in which muscle function further declines.

In addition, muscle function also deteriorates when exercise is insufficient due to the development of transportation means, or when long-term rest is required for medical treatment of illness after surgery. In order to return to daily life sooner after healing, it is desirable to suppress muscle dysfunction.

In order to suppress or recover the decrease in muscle function, it is important to suppress or recover the decrease in muscle mass and muscle quality, and as one of the means for that purpose, exercise therapy such as rehabilitation is performed. On the other hand, from a nutritional point of view, research on components that can improve muscle mass and the like is being conducted. Patent Document 1 describes a muscle mass improving agent containing at least one tyrosine compound and at least one amino acid selected from the group consisting of leucine and isoleucine as active ingredients.

Japanese Unexamined Patent Publication No. 2016-14007

An object of the present invention is to provide a novel composition for suppressing or recovering muscle dysfunction, which can suppress or recover muscle dysfunction. Another object of the present invention is to provide a novel composition for suppressing or recovering muscle mass loss, which can suppress or recover muscle loss. Another object of the present invention is to provide a novel composition for suppressing or recovering muscle loss, which can suppress or recover muscle loss.

As a result of diligent research to solve the above problems, the present inventors have found that a peptide containing the amino acid sequence of His-Lys (histidine-lysine) at the C-terminal or a salt thereof suppresses muscle loss or muscle loss or suppresses muscle loss. It has been found that it has a recovering action and is useful for suppressing or recovering a decrease in muscle function.

That is, the present invention provides the following compositions for suppressing or recovering muscle function, a composition for suppressing or recovering muscle mass loss, and a composition for suppressing or recovering muscle loss.
[1] A composition for suppressing or recovering muscle function decline, which contains a peptide containing the amino acid sequence of His-Lys at the C-terminal or a salt thereof as an active ingredient.
[2] The composition for suppressing or recovering a decrease in muscle function according to the above [1], wherein the decrease in muscle function is a decrease in muscle function due to aging.
[3] A composition for suppressing or recovering muscle mass loss, which contains a peptide containing the amino acid sequence of His-Lys at the C-terminal or a salt thereof as an active ingredient.
[4] The composition for suppressing or recovering muscle mass loss according to the above [3], wherein the muscle mass loss is age-related muscle mass loss.
[5] A composition for suppressing or recovering muscle loss, which contains a peptide containing the amino acid sequence of His-Lys at the C-terminal or a salt thereof as an active ingredient.
[6] The composition for suppressing or recovering muscle loss according to the above [5], wherein the muscle loss is age-related muscle loss.
[7] The composition for suppressing or recovering muscle loss according to the above [5] or [6], wherein the muscle loss is a decrease in adhesion between muscle cells.
[8] The composition according to any one of the above [1] to [7], wherein the peptide containing the amino acid sequence of His-Lys at the C-terminal is any of the following peptides (a) to (c).
(A) X ^1- His-Lys (X ¹ does not exist, or the amino acid at positions n1 to 58 of the amino acid sequence shown in SEQ ID NO: 1 (n1 represents an integer of any of 1 to 58)) Peptide consisting of the amino acid sequence shown in (Representing the sequence) In the amino acid sequence shown in (b) SEQ ID NO: 1, 1 to 9 amino acids were deleted, substituted and / or added in the 1st to 58th regions. Peptide consisting of amino acid sequence (c) Peptide consisting of a partial sequence of the peptide of (b) above and containing the amino acid sequence of His-Lys at the C-terminal [9] The amino acid sequence of His-Lys at the C-terminal. peptides ^{comprising, X 2 -His-Lys (X} 2 is absent or, N2～18 amino acid sequence shown in SEQ ID nO: 2 (n2 represents an integer of 1 to 18) of The composition according to any one of the above [1] to [8], which is a peptide consisting of the amino acid sequence shown in (representing the amino acid sequence).
[10] The peptide containing the amino acid sequence of His-Lys at the C-terminal is a peptide consisting of the amino acid sequence of His-Lys, Leu-His-Lys or Leu-Leu-His-Lys (SEQ ID NO: 3). 1] The composition according to any one of [9].
[11] The composition according to any one of the above [1] to [10], which is an oral composition.

According to the present invention, there is provided a novel composition for suppressing or recovering muscle dysfunction, which can suppress or recover muscle dysfunction. Further, according to the present invention, there is provided a novel composition for suppressing or recovering muscle mass loss, which can suppress or recover muscle loss. According to the present invention, there is provided a novel composition for suppressing or recovering muscle loss, which can suppress or recover muscle loss.

1A-E are photomicrographs of HE-stained rat skeletal muscle (scale bar: 50 μm).

The composition for suppressing or recovering muscle function deterioration of the present invention contains a peptide or a salt thereof containing the amino acid sequence of His-Lys at the C-terminal as an active ingredient.
The composition for suppressing or recovering muscle loss of the present invention contains a peptide containing the amino acid sequence of His-Lys at the C-terminal or a salt thereof as an active ingredient.
The composition for suppressing or recovering muscle loss of the present invention contains a peptide or a salt thereof containing the amino acid sequence of His-Lys at the C-terminal as an active ingredient.

In the present specification, a peptide containing the amino acid sequence of His-Lys at the C-terminal is also described as an HK-containing peptide. One kind of HK-containing peptide or a salt thereof may be used, or two or more kinds may be used.
The salt of the peptide is not particularly limited as long as it is a pharmacologically acceptable salt, and may be either an acidic salt or a basic salt. As acidic salts, for example, inorganic acid salts such as hydrochlorides, sulfates, nitrates, phosphates; acetates, citrates, maleates, malates, oxalates, lactates, succinates, fumals. Examples thereof include organic acid salts such as acid salts and propionates. Examples of the basic salt include alkali metal salts such as sodium salt and potassium salt; and alkaline earth metal salts such as calcium salt and magnesium salt.

The number of amino acid residues of the peptide containing the amino acid sequence of His-Lys at the C-terminal may be, for example, 2 to 100. The peptide containing the His-Lys amino acid sequence at the C-terminal may be a peptide consisting of the His-Lys amino acid sequence.
As the peptide containing the amino acid sequence of His-Lys at the C-terminal, the peptides (a) to (c) below are preferable.
(A) X ^1- His-Lys (X ¹ does not exist, or the amino acid at positions n1 to 58 of the amino acid sequence shown in SEQ ID NO: 1 (n1 represents an integer of any of 1 to 58)) Peptide consisting of the amino acid sequence shown in (Representing the sequence) (b) In the amino acid sequence shown in SEQ ID NO: 1, 1 to 9 amino acids were deleted, substituted and / or added in the 1st to 58th regions. Peptide consisting of amino acid sequence (c) The amino acid sequence shown in Peptide SEQ ID NO: 1, which is a peptide consisting of a partial sequence of the peptide of (b) above and contains the amino acid sequence of His-Lys at the C-terminal, is that of a jersey cattle. It is the 1st to 60th amino acid sequence from the amino end of β-lactoglobulin C.

In the present specification, deletion, substitution and / or addition of 1 to 9 amino acids in an amino acid sequence means that 1 to 9 amino acids are deleted, substituted and / or added at any position in the amino acid sequence of 1 to 9 in the same sequence. It means that there is an amino acid deletion, substitution and / or addition, and two or more of the deletion, substitution and addition may occur at the same time.

In the amino acid sequence shown in SEQ ID NO: 1, the peptide (b) is preferably 1 to 8 (more preferably 1 to 7, 1 to 6, 1 to 5) in the 1 to 58th regions. Or it consists of a sequence in which 1 to 4, more preferably 1 to 3, 1 to 2 or 1) amino acids are deleted, substituted and / or added.
The peptide of (c) above is a peptide consisting of a partial sequence of the peptide according to (b) above and containing an amino acid sequence of His-Lys at the C-terminal.

The peptide of (a) above is more preferable as the peptide containing the amino acid sequence of His-Lys at the C-terminal.
In one embodiment, as the peptide of (a), X ^2- His-Lys (X ² is absent, or n2 is any of 1 to 18 of the amino acid sequence shown in SEQ ID NO: 2). A peptide consisting of the amino acid sequence represented by) (representing the integer) is preferable. The amino acid sequence shown in SEQ ID NO: 2 is the 41st to 60th amino acid sequence of the amino acid sequence shown in SEQ ID NO: 1 (the peptide in which n1 represents an integer of 41 to 58 in the peptide of (a)).

As the peptide containing the amino acid sequence of His-Lys at the C-terminal, a peptide having the amino acid sequences of His-Lys, Leu-His-Lys, and Leu-Leu-His-Lys (SEQ ID NO: 3) is more preferable. These peptides or salts thereof are preferable from the viewpoints of suppressing or recovering muscle function, suppressing or recovering muscle mass loss, and suppressing or recovering muscle loss. As a peptide or a salt thereof containing the amino acid sequence of His-Lys at the C-terminal, a peptide consisting of the amino acid sequence of His-Lys, Leu-His-Lys or Leu-Leu-His-Lys (SEQ ID NO: 3) or a salt thereof is further added. Preferably, a peptide consisting of the amino acid sequence of Leu-His-Lys or a salt thereof is particularly preferable.

The method for producing a peptide or a salt thereof containing the amino acid sequence of His-Lys at the C-terminal is not particularly limited. The HK-containing peptide can be produced, for example, by a known peptide synthesis method. When the HK-containing peptide is obtained by chemical synthesis, it can be synthesized by either a solid-phase method or a liquid-phase method. The peptide obtained by the synthesis can be purified by a usual purification method using reverse phase high performance liquid chromatography, ion exchange resin, chromatography using a high porous polymer resin, affinity chromatography and the like. Further, for example, His-Lys, Leu-His-Lys and the amino acid sequence shown in SEQ ID NO: 3 (Leu-Leu-His-Lys) are contained in β-lactoglobulin C contained in the milk of Jersey cows. It is a sequence and can also be produced by degrading β-lactoglobulin C with an enzyme or the like. Peptides containing the His-Lys amino acid sequence at the C-terminus can also be biosynthesized using microorganisms. In the case of biosynthesis using a microorganism, an expression vector in which a gene encoding a peptide containing a His-Lys amino acid sequence is introduced at the C-terminal is prepared, and the expression vector is introduced into a host microorganism to prepare a transformant. obtain. The transformant can be cultured and a peptide containing the His-Lys amino acid sequence at the C-terminus can be purified from the culture. The host microorganism is not particularly limited, and for example, Escherichia coli, yeast and the like can be used. Separation and purification from the culture can be performed by a conventional purification method.
Peptide salts can be readily prepared by one of ordinary skill in the art by any method known in the art.

As used herein, suppressing or recovering a decrease in muscle function refers to suppressing the decrease in muscle function or restoring the decreased muscle function. Muscle function is a concept that includes muscle strength, elastic force, endurance, and the like. The muscle dysfunction may be an age-related muscle dysfunction.
Suppression or recovery of muscle mass loss refers to suppressing or recovering muscle mass loss. Suppression of muscle loss is a concept that includes muscle mass gain. The muscle loss may be age-related muscle loss.
Suppression or recovery of muscle loss refers to suppressing loss of muscle (muscle condition) or recovering weakened muscle. The muscle loss may be age-related muscle loss. Examples of muscle deterioration include a decrease in adhesion between muscle cells. When the adhesion between muscle cells is reduced, muscle function is reduced.
Suppressing or restoring the decrease in adhesion between muscle cells refers to suppressing the decrease in adhesion between muscle cells or restoring the decreased adhesion between muscle cells. Suppression of reduced adhesion between muscle cells is a concept that includes strengthening adhesion between muscle cells.
The muscle mass of animals such as humans can be measured by, for example, microPET / CT (positron emission tomography / computed tomography, INVEON, Siemens, USA).

As shown in the examples below, aged rats had reduced adhesion between myocytes in skeletal muscle compared to young rats. When aged rats ingest a peptide containing the amino acid sequence of His-Lys or a salt thereof at the C-terminal, the number of muscle cells is increased and the adhesion between the muscle cells is strengthened as compared with the control aged rat. It was suggested. In addition, rats fed with a peptide containing the amino acid sequence of His-Lys or a salt thereof had more skeletal muscle mass than rats not fed. Therefore, a peptide or a salt thereof containing the His-Lys amino acid sequence at the C-terminal is useful for suppressing or recovering muscle loss. In addition, a peptide containing the His-Lys amino acid sequence at the C-terminal or a salt thereof suppresses or restores the decrease in adhesion between muscle cells, and is therefore useful for suppressing or recovering the decrease in muscle quality. By suppressing or recovering muscle mass loss or suppressing or recovering muscle quality loss, it is possible to suppress or recover muscle function loss. In one embodiment, a peptide or salt thereof containing the His-Lys amino acid sequence at the C-terminus is suitable for suppressing or recovering muscle dysfunction by suppressing or recovering muscle mass loss and / or muscle loss. Can be used.

A peptide or a salt thereof containing the His-Lys amino acid sequence at the C-terminal is useful for suppressing or recovering muscle dysfunction, for suppressing or recovering muscle loss, or for suppressing or recovering muscle loss. .. Peptides or salts thereof containing the amino acid sequence of His-Lys at the C-terminal are used to suppress or restore skeletal muscle dysfunction, to suppress or restore skeletal muscle mass loss, and to reduce skeletal muscle muscularity. Useful for suppression or recovery. In one aspect of the present invention, the peptide or salt thereof containing the amino acid sequence of His-Lys at the C-terminal is used for suppressing or recovering muscle function decline due to aging, and for suppressing or recovering muscle mass loss due to aging. It is suitably used for suppressing or recovering muscle deterioration due to aging.
The use of the above peptides or salts thereof may be therapeutic or non-therapeutic. "Non-therapeutic" is a concept that does not include medical practice, ie surgery, treatment or diagnosis.

The composition for suppressing or recovering muscle dysfunction, the composition for suppressing or recovering muscle mass loss, and the composition for suppressing or recovering muscle loss of the present invention are collectively referred to below as the composition for suppressing or recovering muscle dysfunction. Also called a thing.
The composition for suppressing or recovering muscle function decline of the present invention can be provided, for example, in the form of food or drink (food or drink composition), pharmaceutical product (pharmaceutical composition), feed (feed composition) or the like. , Not limited to these. The composition for suppressing or recovering muscle function deterioration of the present invention may itself be a food or drink, a pharmaceutical product, a feed or the like, or may be a preparation or a material such as an additive used for these. In one aspect, from the viewpoint of more fully exerting the effects of the present invention, the composition for suppressing or recovering muscle function decline is preferably an oral composition, and is preferably a food or drink or an oral drug. More preferably, it is a food or drink.

The composition for suppressing or recovering muscle function deterioration of the present invention contains any additive or any component in addition to the HK-containing peptide or a salt thereof used as an active ingredient as long as the effect of the present invention is not impaired. Can be contained. As these additives and ingredients, those that can be generally used for foods and drinks, pharmaceuticals, feeds and the like can be used.
The composition for suppressing or recovering muscle function deterioration of the present invention can be provided in the form of an agent as an example, but is not limited to this form. The agent can be provided as it is as a composition or as a composition containing the agent.

When the composition for suppressing or recovering muscle function deterioration of the present invention is used as a food or drink, a component that can be used in the food or drink (for example, a food or drink material, if necessary) is used in the HK-containing peptide or a salt thereof. Food additives, etc.) can be blended to make various foods and drinks. Foods and drinks are not particularly limited, and examples thereof include general foods and drinks, health foods, and raw materials thereof. The form of the food or drink is not particularly limited, and examples thereof include solid, semi-fluid, and fluid. Foods and drinks are oral solid preparations such as tablets, coated tablets, fine granules, granules, powders, pills, capsules, dry syrups and chewables; various forms of oral liquid preparations such as oral liquids and syrups. It can also be.

When the composition for suppressing or recovering muscle function deterioration of the present invention is used as a pharmaceutical product, a pharmacologically acceptable excipient or the like is mixed with the HK-containing peptide or a salt thereof to obtain various dosage forms of the pharmaceutical product. can do. The administration form of the drug is not particularly limited, and may be oral administration or parenteral administration. From the viewpoint of obtaining the effect of the present invention more sufficiently, the administration form of the drug is preferably oral administration. The dosage form of the drug may be a dosage form suitable for the dosage form. Dosage forms of oral pharmaceuticals include, for example, oral solid preparations such as tablets, coated tablets, fine granules, granules, powders, pills, capsules, dry syrups, chewables; oral solutions such as oral liquids and syrups. Examples include liquid preparations. Dosage forms of pharmaceuticals for parenteral administration include injections, infusions, ointments, lotions, patches, suppositories, nasal agents, pulmonary agents (inhalants) and the like. The drug may be a non-human animal drug.

When the composition for suppressing or recovering muscle function deterioration of the present invention is used as a feed, a component that can be used in the feed can be mixed with the HK-containing peptide or a salt thereof to prepare the feed. Examples of the feed include livestock feed used for cattle, pigs, chickens, sheep, horses, etc .; small animal feed used for rabbits, guinea pigs, rats, mice, etc .; pet food used for dogs, cats, small birds, etc. ..
When the composition for suppressing or recovering muscle function deterioration of the present invention is a food or drink, a pharmaceutical product, a feed or the like, the production method thereof is not particularly limited. For example, it can be produced by a general method using an HK-containing peptide or a salt thereof used as an active ingredient.

The content of the HK-containing peptide or a salt thereof in the composition for suppressing or recovering muscle function deterioration of the present invention is not particularly limited and can be appropriately set according to the form and the like. For example, when a composition for suppressing or recovering muscle function decline is used as a food or drink, a pharmaceutical product, a feed, etc., the total content of the HK-containing peptide or its salt (the HK-containing peptide and its salt) in any form. The total content) may be, for example, 0.001% by weight or more, preferably 0.01% by weight or more, 90% by weight or less, and preferably 80% by weight or less in the composition. In one embodiment, the total content of the HK-containing peptide or salt thereof may be, for example, 0.001 to 90% by weight, preferably 0.01 to 90% by weight, in the composition for suppressing or recovering muscle function decline. It can be 80% by weight.

The composition for suppressing or recovering muscle function deterioration of the present invention can be administered or ingested by an appropriate method according to its form, and is preferably administered or ingested orally.
The intake amount (which can also be referred to as a dose) of the composition for suppressing or recovering muscle function deterioration of the present invention is not particularly limited, and suppresses or recovers muscle loss, suppresses or recovers muscle loss, and muscles. The amount may be any amount (effective amount) that can suppress or recover the functional deterioration, and may be appropriately set according to the administration form, administration method, and the like. In one embodiment, when a human (adult) is orally administered or ingested, the total intake of the HK-containing peptide or its salt (total intake of the HK-containing peptide and its salt) is the C-terminus of the HK-containing peptide. In terms of the weight of His-Lys, for example, at a body weight of 50 kg per day, 0.1 mg or more is preferable, 1.0 mg or more is more preferable, 2.5 mg or more is further preferable, and 10 mg or more is particularly preferable. , 2.5 g or less, more preferably 1000 mg or less, further preferably 500 mg or less, and particularly preferably 100 mg or less. In one embodiment, the total intake of the HK-containing peptide or salt thereof in the case of a human (adult) is 50 kg in body weight per day, and is 0.1 mg in terms of the weight of His-Lys at the C-terminal of the HK-containing peptide. ~ 2.5 g is preferable, 1.0 to 1000 mg is more preferable, 2.5 to 500 mg is further preferable, 10 to 500 mg is even more preferable, and 10 to 100 mg is particularly preferable. It is preferable that the above amount is orally administered or ingested, for example, once a day or in a plurality of times (for example, 2 to 3 times). It is preferable to administer or ingest the composition for suppressing or recovering muscle function decline of the present invention or the like so that the intake amount of the HK-containing peptide or a salt thereof is within the above range. The composition for suppressing or recovering muscle function deterioration of the present invention is an oral composition for administering or ingesting the above amount of HK-containing peptide or salt thereof per day per 50 kg of body weight to humans. Good.

The composition for suppressing or recovering muscle function deterioration of the present invention is preferably continuously ingested (administered). When the HK-containing peptide or its salt is continuously ingested, the effect of suppressing or recovering muscle function, the effect of suppressing or recovering muscle mass loss, and the effect of suppressing or recovering muscle loss are enhanced. In one embodiment of the present invention, the composition for suppressing or recovering muscle function decline is preferably ingested continuously for 2 weeks or more, and more preferably continuously for 4 weeks or more.

As described above, the HK-containing peptide or a salt thereof has an inhibitory or recovering effect on muscle function decline, an inhibitory or recovering effect on muscle mass loss, and a suppressing or recovering effect on muscle loss. For this reason, HK-containing peptides or salts thereof are useful for preventing or ameliorating conditions or diseases caused by muscle dysfunction, muscle loss or muscle loss.
Conditions or diseases caused by decreased muscle function, decreased muscle mass, or decreased muscle quality include sarcopenia, bedridden, falls, tiredness, and decreased QOL. In one aspect, the composition for suppressing or recovering muscle function decline of the present invention can be used for prevention or improvement of the above-mentioned conditions or diseases. Prevention includes prevention of onset, delay of onset, and reduction of incidence. Improvement includes symptom relief, symptom progression suppression, and symptom healing.

The subject to which the composition for suppressing or recovering muscle function deterioration of the present invention is administered or ingested (hereinafter, also simply referred to as an administration subject) is preferably a human or a non-human animal, and more preferably a human or a non-human mammal. Humans are even more preferred. Examples of non-human mammals include cows, pigs, sheep, horses, rabbits, dogs, cats, guinea pigs, rats, mice, monkeys and the like. In addition, as a target for administration of a composition for suppressing or recovering muscle dysfunction, a subject who needs or desires suppression or recovery of muscle dysfunction, a subject who needs or desires suppression or recovery of muscle mass loss, a muscle deterioration Subjects who need or desire to suppress or recover from are preferred. For example, a subject who desires prevention or improvement of a condition or disease caused by the above-mentioned decrease in muscle function, decrease in muscle mass, or decrease in muscle quality can be mentioned.
Since muscle function, muscle mass, and muscle quality decrease with age, as an example of a preferable administration target in the present invention, a subject who needs or desires suppression or recovery of the decrease in muscle function, muscle mass, or muscle quality due to age. Can be mentioned. In one aspect, the elderly are preferred as administration targets.

The composition for suppressing or recovering muscle function deterioration of the present invention includes one of the uses, types of active ingredients, the above-mentioned effects, usage methods (for example, ingestion method, administration method), etc. in packaging, containers or instructions. Two or more may be displayed. The composition for suppressing or recovering muscle function deterioration of the present invention is labeled as having an action of suppressing or recovering muscle function loss, muscle mass loss, muscle loss, or an action based on this action. You may be.

In one aspect, the composition for suppressing or recovering muscle function decline of the present invention includes "maintenance of walking ability (function)", "maintenance of walking function", "maintenance of muscle mass", and "improvement of muscle mass". , "Suppression of muscle weakness", "Supporting muscle building power", etc. may be labeled with one or more. In one embodiment of the present invention, the composition for suppressing or recovering muscle function decline of the present invention is preferably a food or drink with the above-mentioned function indication. Further, the above display may be a display to the effect that it is used to obtain the above function.

The present invention also includes the following methods.
A method for suppressing or recovering muscle dysfunction, which comprises administering to a subject a peptide or a salt thereof containing the amino acid sequence of His-Lys at the C-terminal.
A method for suppressing or recovering muscle loss, which comprises administering to a subject a peptide or a salt thereof containing the amino acid sequence of His-Lys at the C-terminal.
A method for suppressing or recovering muscle loss, which comprises administering to a subject a peptide or a salt thereof containing the amino acid sequence of His-Lys at the C-terminal.
Peptides containing the amino acid sequence of His-Lys at the C-terminal (HK-containing peptide) or salts thereof, administration targets, administration methods, dosages and preferred embodiments thereof include the above-mentioned compositions for suppressing or recovering muscle function deterioration. Is the same as. The dose of the HK-containing peptide or a salt thereof may be an amount that can suppress or recover muscle function loss, suppress or recover muscle loss, suppress or recover muscle loss, that is, an effective amount. There is no particular limitation. For example, it is preferable to administer the above-mentioned intake amount. The HK-containing peptide or salt thereof may be administered as it is, or may be administered as a composition containing the HK-containing peptide or salt thereof. For example, the above-mentioned composition for suppressing or recovering muscle function decline of the present invention can be administered. According to the present invention, muscle mass loss and muscle loss can be suppressed or recovered. In addition, it is possible to suppress or recover the decrease in muscle function. In addition, it is possible to prevent or ameliorate a condition or disease caused by a decrease in muscle function, a decrease in muscle mass, or a decrease in muscle quality. The above method may be a therapeutic method or a non-therapeutic method. In one aspect, the muscle dysfunction may be age-related muscle dysfunction. The muscle loss may be age-related muscle loss. The muscle loss may be age-related muscle loss.

The present invention also includes the following uses.
Use of a peptide or salt thereof containing the His-Lys amino acid sequence at the C-terminus to suppress or restore muscle dysfunction.
Use of peptides or salts thereof containing the His-Lys amino acid sequence at the C-terminus to suppress or restore muscle loss.
Use of a peptide or salt thereof containing the His-Lys amino acid sequence at the C-terminus to suppress or restore muscle loss.
The above use is for human or non-human animals, preferably for humans. The use may be therapeutic or non-therapeutic use. The HK-containing peptide or salt thereof, the subject, the administration method, the dose, and preferred embodiments thereof are the same as those in the above-mentioned composition for suppressing or recovering muscle function decline.

The present invention uses a peptide containing the amino acid sequence of His-Lys at the C-terminal or a salt thereof for producing a composition for suppressing or recovering muscle loss; producing a composition for suppressing or recovering muscle loss. Use of a peptide containing the amino acid sequence of His-Lys at the C-terminal or a salt thereof; a peptide containing the amino acid sequence of His-Lys at the C-terminal for producing a composition for suppressing or recovering muscle loss. Or the use of its salts. The composition for suppressing or recovering muscle function, the composition for suppressing or recovering muscle mass loss, the composition for suppressing or recovering muscle loss, and preferred embodiments thereof are the same as described above.

Hereinafter, examples will be shown in which the present invention will be described in more detail. The present invention is not limited to these examples.

A series of animal experiments were conducted based on a plan approved by the head of the institution after being examined by the in-house and on-campus animal experiment committees in compliance with the Act on Welfare and Management of Animals and other related laws and regulations.

<Example 1>
Experimental method 1. As experimental animal old rats, Wister male rats were purchased at 5 months of age (Nippon SLC Co., Ltd.) and subjected to the experiment at 17 months of age. Young rats were purchased at 2 months of age and used at 3 months of age.

2. Test substance The following test substance was used.
Acetate of a tetrapeptide consisting of the amino acid sequence of Leu-Leu-His-Lys (SEQ ID NO: 3) (hereinafter referred to as LLHK).
Acetate of a tripeptide consisting of the amino acid sequence of Leu-His-Lys (hereinafter referred to as LHK)
A salt of a dipeptide consisting of the amino acid sequence of His-Lys (hereinafter referred to as HK).
The synthesis of LLHK (acetate) and LHK (acetate) was commissioned to Peptide Institute of Japan (Minoh City).

3. 3. The test substance was dissolved in tap water so that the drinking LLHK was 30 mg / 30 mL, the LHK was 23 mg / 30 mL, and the HK was 16 mg / 30 mL, and the rats were allowed to drink. The His-Lys concentration in the HK aqueous solution was 12 mg / 30 mL.
17-month-old rats were fed tap water (water drinking group, old-age control), LLHK (LLHK drinking group), LHK (LHK drinking group) or HK (HK drinking group) to ad lib and bred for 1 month ( Each group n = 2). Three-month-old rats (young control) were fed tap water and bred for one month (n = 2). During this experiment, the same solid feed (trade name MF, manufactured by Oriental Yeast Co., Ltd.) was freely used in the test substance (LLHK, LHK or HK) drinking group, water drinking group (control) and young control group. Was ingested. Rats ingested about 22 mL of the above aqueous test substance solution daily.
After the above breeding (after the end of the experiment), the weight of the rats in each group was measured. Then, the following test was conducted.

4. Measurement of muscle weight and analysis of histology For the LLHK drinking group, HK drinking group and young control group, rat skeletal muscle (soleus muscle) was excised and immediately 10% neutral buffered formaldehyde solution (manufactured by Nacalai Tesque, Inc.). After fixing with, a 5 mm section was prepared. After staining with hematoxylene and eosin (HE staining), it was observed with an optical microscope (BX53 manufactured by Olympus Corporation) (magnification 10 × 20).
For the water drinking group and the LHK drinking group, rat skeletal muscle (soleus muscle) was excised and immediately weighed. After the weight measurement, the skeletal muscle was fixed by the same method as described above and observed with an optical microscope.

5. Microarray analysis The excised skeletal muscle is immediately placed in an RNA lattice (Ambion, Austin, TX, USA), and total RNA purification is performed using RNeasy Fibrow Tissue Mini Kit (Qiagen GmbH, Hilden, Germany) and QI (Germany). went. RNA purity was confirmed using NanoDrop 1000 spectrophotometer (Thermo Fisher Scientific Inc., Wilmington, DE, USA) and Agilent 2100 Bioanalyzer (Agilent Technologies, USA), Agilent Technologies, and Clara, USA. Labeled cRNA was synthesized and fragmented using (Agilent Technologies, Santa Clara, Ca, USA). Hybridization was performed at SurePrint G3 Rat GE ver 2.0 (Agilent Technologies) and 2545A Hybridization Oven (Agilent Technologies) at 65 ° C. for 17 hours. This Array can analyze about 30,000 genes. GeneSpring GX version 14.9 (Agilent Technologies) was used for the analysis.

(result)
The values obtained are shown as mean ± standard error. The significance test is the test result for the drinking group (old age control) performed by t-test (test with two samples assuming homoscedasticity, p-value, one-sided test) using Excel's analysis tool (*). *: P <0.01).

1. 1. Effect of test substance on body weight Table 1 shows the body weight of rats before the start of the experiment and after drinking the test substance (LLHK, LHK or HK) or water for one month (after the end of the experiment). The body weight tended to decrease with aging in all of the water drinking group (aging control), the LLHK drinking group, the LHK drinking group, and the HK drinking group, but there was no significant difference between the groups (Table 1).

2. Effect of Test Substance on Tissue Image of Skeletal Muscle Figures 1A to 1E are micrographs of HE-stained rat skeletal muscle (scale bar: 50 μm). 1A to 1E show A is a young control (drinking water), B is an old age control (drinking water), C is a rat that drank LLHK, D is a rat that drank LHK, and E is a rat that drank HK. .. In young animals, myocytes are neatly bundled (Fig. 1A). However, in the aged animals for drinking water, the bonds between the muscle cells were broken and separated (Fig. 1B). When LLHK is drunk (Fig. 1C), LHK is drunk (Fig. 1D), and HK is drunk (Fig. 1E), the muscle cells recover their binding force and become lumpy, approaching the image of a young animal (Fig. 1A). It was.

3. 3. Effect of test substance on muscle weight Table 2 shows the weight of rat soleus muscle after drinking LHK or water for one month (after the end of the experiment). In the LHK drinking group, the muscle weight was significantly heavier than in the water drinking group (aging control) (**: p <0.01, n = 2).

4. Effect of test substance on gene expression of skeletal muscle We analyzed genes in which LLHK, LHK, or HK was increased by 1.5 times or more compared to drinking water, and genes in which young control was higher than old control. Drinking LLHK, LHK or HK increased 1139 entities as a common gene. For genes that were 1.5 times or more lower than drinking water when drinking LLHK, LHK, or HK, and genes that were lower in the young control than in the old control, 1289 genes were reduced as common genes.

Genes were examined by function (Tables 3-8).
Columns A in Tables 3 to 8 are values (OLLHK / OC) obtained by dividing the gene expression level (OLLHK) of LLHK drinking aged animals (LLHK drinking group) by the gene expression level (OC) of water drinking aged animals (aging control). is there. Column B is a value (OLHK / OC) obtained by dividing the gene expression level (OLHK) of the LHK drinking aged animals (LHK drinking group) by the gene expression level (OC) of the water drinking aged animals. Column C is a value (OHK / OC) obtained by dividing the gene expression level (OHK) of the HK drinking old animals (HK drinking group) by the gene expression level (OC) of the water drinking old animals. Column D is a value (YC / OC) obtained by dividing the gene expression level (YC) of a drinking young animal (young control) by the gene expression level (OC) of a drinking water old animal.
In Tables 3-5 and 7-8 below, the values shown in columns A, B and C that are closest to the values in column D (YC / OC) are underlined. This underlined number means that the gene expression of each group is closest to that of the young control. In the results shown in Tables 3-5 and 7-8, the gene expression of the animals that drank LHK was the closest to the gene expression of the young animals.

Table 3 shows the results of examining the expression of genes that promote muscle development (promote muscle mass increase). Insulin-like growth factor genes (Igf1), insulin-like growth factor-binding protein genes (Igf2bp2, Igfbp6, Igfbp4), and insulin-like growth factor receptor genes (Igf1) and insulin-like growth factor receptor genes (Igf1) and insulin-like growth factor-binding protein genes (Igf2bp2, Igfbp6, Igfbp4) are used as genes that promote muscle development by LLHK, LHK, or HK. The expression of Igf1r), transcriptional prolonging factor (Tceanc2), and ankirin 44 (Ankdr44), which is a protein lining the cell membrane, was increased.

Table 4 shows the results of examining the expression of genes related to cell adhesion. By LLHK, LHK or HK, as genes related to cell adhesion, Nectin gene (Nectin3, Fndc1), Hyaluronic acid receptor gene (Cd44), Integrin gene (Itga2, Itga3, Itgb1, Itgb7, Itfg2), Cadherin Expression of genes (Cad4, Pcdha5, Pcdh17) and the like was increased.

Table 5 shows the results of examining the expression of genes involved in muscle repair. LLHK, LHK, or HK increased the expression of chemokine ligand genes (Ccl7, Cxcl9, Cxcl16), catenin (Ctnnbip1, Ctnna2), etc. as genes involved in cell repair.

Table 6 shows the results of examining the expression of genes that promote differentiation into skeletal muscle cells. LLHK, LHK or HK increased the expression of Pax3 as a gene that promotes differentiation into skeletal muscle cells. The higher the expression level of the gene that promotes differentiation into skeletal muscle cells, the more the differentiation into skeletal muscle is promoted and the skeletal muscle mass increases.

Table 7 shows the results of examining the expression of genes involved in cell matrix remodeling. LLHK, LHK or HK increased the expression of CTGF and Dcn as genes involved in cell matrix remodeling.

Table 8 shows the results of examining the expression of genes that promote the growth of skeletal muscle. By LLHK, LHK or HK, the expression of growth factor genes (Ltbp1, Tgfb2, Tgfbi, Ltbp4) and the like as genes promoting the growth of skeletal muscle was increased.

The results shown in FIG. 1 show that LLHK, LHK, and HK promoted morphogenesis by enhancing the adhesion of disjointed cells due to poor cell adhesion in old age. In FIG. 1B, the adhesion between muscle cells is loose, and the number of cells in one muscle bundle is smaller than that in FIG. 1A, whereas in FIGS. 1C to E (particularly, FIGS. 1C and 1D), between muscle cells. The adhesion of the cells became stronger, and the number of cells in one muscle bundle increased from that in FIG. 1B. When the adhesion between muscle cells is strengthened, muscle function is improved. It is considered that the enhancement of cell adhesion and morphogenesis by LLHK, LHK, and HK is due to the increase in gene expression related to cell adhesion and muscle repair shown in Tables 3 to 8. For example, as shown in Table 4, LHK, LHK, and HK increase the expression of genes involved in muscle cell adhesion. The enhancement of adhesion between muscle cells by LHK, LHK, and HK is considered to be due to the expression of these genes. In addition, as shown in Table 2, the rats that drank LHK had significantly heavier muscle weight than the rats that did not drink LHK.

As can be seen in Table 3, drinking LLHK, LHK, and HK increases various insulin-like growth factor gene (Igf) expression, insulin-like growth factor binding protein gene, and insulin-like growth factor receptor gene gene. I let you. There are many reports that insulin-like growth factor increases muscle mass. For example, insulin / insulin-like growth factor activates the PI (3) K / Akt signaling system to increase muscle protein synthesis and thicken myotubes (Rommel C. Et al, Nature Cell Biol 3). : 1009-1013, 2001).
In addition, as shown in Table 5, the expression of catenin (Ctnana2) was increased by LLHK, LHK, and HK. Catenin forms a complex with cadherin and is essential for cadherin to exert its function as a cell adhesion factor, and also has a function as a transcriptional regulator (Valenta T, et al., The EMBO Journal 31 (12): 2714). -36, 2012.). From these results, drinking a peptide consisting of the amino acid sequence of His-Lys, Leu-His-Lys or Leu-Leu-His-Lys (SEQ ID NO: 3) or a salt thereof is effective in suppressing or recovering muscle loss. It can be said that there is. In addition, it can be said that drinking the peptide or a salt thereof is effective in suppressing or recovering muscle deterioration. Therefore, the peptide or a salt thereof is effective for sarcopenia (age-related muscle weakness) and the like.

HK-containing peptides such as peptides consisting of the amino acid sequences of His-Lys, Leu-His-Lys or Leu-Leu-His-Lys (SEQ ID NO: 3) or salts thereof cause senile tissue changes in skeletal muscle and deterioration of senile function. It turned out to be effective for repair.
Further, as described above, the HK-containing peptide or a salt thereof has an effect of strengthening the adhesion between muscle cells. In recent years, research on the production of artificial meat by culturing has been active, and the addition of an HK-containing peptide or a salt thereof can be expected to have an effect of promoting artificial meat formation. In particular, Leu-His-Lys or its salt does not rely on animal-derived meat, which is likely to have the ability to clump loose meat cells, and when making artificial meat in a factory or the like, Leu-His-Lys HK-containing peptides such as, or salts thereof may be one of the important tools for food supply.

Claims (11)

A composition for suppressing or recovering muscle dysfunction, which contains a peptide containing the amino acid sequence of His-Lys at the C-terminal or a salt thereof as an active ingredient. The composition for suppressing or recovering a decrease in muscle function according to claim 1, wherein the decrease in muscle function is a decrease in muscle function due to aging. A composition for suppressing or recovering muscle mass loss, which contains a peptide containing the amino acid sequence of His-Lys at the C-terminal or a salt thereof as an active ingredient. The composition for suppressing or recovering muscle mass loss according to claim 3, wherein the muscle mass loss is age-related muscle mass loss. A composition for suppressing or recovering muscle loss, which contains a peptide containing the amino acid sequence of His-Lys at the C-terminal or a salt thereof as an active ingredient. The composition for suppressing or recovering muscle loss according to claim 5, wherein the muscle loss is age-related loss of muscle strength. The composition for suppressing or recovering muscle loss according to claim 5 or 6, wherein the muscle loss is a decrease in adhesion between muscle cells. The composition according to any one of claims 1 to 7, wherein the peptide containing the amino acid sequence of His-Lys at the C-terminal is any of the following peptides (a) to (c).
(A) X ^1- His-Lys (X ¹ does not exist, or the amino acid at positions n1 to 58 of the amino acid sequence shown in SEQ ID NO: 1 (n1 represents an integer of any of 1 to 58)) Peptide consisting of the amino acid sequence shown in (Representing the sequence) (b) In the amino acid sequence shown in SEQ ID NO: 1, 1 to 9 amino acids were deleted, substituted and / or added in the 1st to 58th regions. Peptide consisting of amino acid sequence (c) Peptide consisting of a partial sequence of the peptide of (b) above, which contains the amino acid sequence of His-Lys at the C-terminal. Said peptide comprising the amino acid sequence of His-Lys at the ^{C-terminus, X 2 -His-Lys (X} 2 is absent, or, n2~18 th (n2 of the amino acid sequence shown in SEQ ID NO: 2, 1 The composition according to any one of claims 1 to 8, which is a peptide consisting of the amino acid sequence represented by) (representing the amino acid sequence of any of 18 to 18). The peptide containing the amino acid sequence of His-Lys at the C-terminal is a peptide consisting of the amino acid sequence of His-Lys, Leu-His-Lys or Leu-Leu-His-Lys (SEQ ID NO: 3). The composition according to any one of the above. The composition according to any one of claims 1 to 10, which is an oral composition.