JP2024117078A - Creatine-containing composition - Google Patents

Abstract

The present invention provides a composition for promoting creatine storage. The composition contains (a) a creatine compound, (b) a naturally occurring component that exhibits a yellow color, and (c) isoleucine or a salt thereof. [Selected Figure]

Description

The present invention relates to a composition that is effective in promoting creatine storage.

The energy required for vital activities such as moving the body or maintaining its functions comes from adenosine triphosphate (ATP). Therefore, continuous supply of this ATP is essential for continuous exercise. In particular, during intense exercise, a large amount of ATP is used, making it necessary to resynthesize ATP. A substance in the body that has the function of resynthesizing ATP is a high-energy phosphate compound. One representative high-energy phosphate compound is creatine phosphate. Creatine phosphate reacts with adenosine diphosphate (ADP) to restore the amount of ATP, thereby supplying energy, so creatine phosphate is an important energy storage substance for physical activity. Creatine is a component of creatine phosphate, and when creatine increases in the body, creatine phosphate also increases. Therefore, by increasing the amount of creatine stored in the body tissues, the amount of energy stored in the body also increases. It is also known that oral intake of creatine compounds increases the amount of free creatine and creatine phosphate in the body tissues (Non-Patent Documents 1 and 2).

Creatine compounds ingested from food are taken up by various organs, including skeletal muscles. Ingestion of creatine compounds not only increases the amount of creatine in muscle tissue, but also increases the amount of creatine in brain tissue, promoting energy storage in the brain tissue. Creatine has been used by athletes and training enthusiasts as a food to improve athletic function and increase muscle mass (Non-Patent Document 3).

On the other hand, in order to increase the amount of creatine stored in body tissues, it may be necessary to ingest a large amount of a creatine compound, and compositions and methods have been proposed for increasing the amount of creatine in tissues in order to reduce the amount of ingested creatine.
For example, simultaneous intake of creatine with sugars used as energy increases the blood insulin concentration. Insulin acts on cells in various tissues to increase the activity of creatine transporters. This increases the amount of creatine taken up into cells, resulting in an increase in the amount of creatine stored in tissues.
Similarly, α-lipoic acid is known to activate the insulin cascade and increase the amount of creatine taken up when used in combination with creatine (Patent Document 1, Non-Patent Document 4). However, when people with a certain genetic predisposition take α-lipoic acid, they may experience symptoms of insulin autoimmune syndrome, such as trembling of the hands and feet and cold sweats, due to hypoglycemic conditions, and therefore caution is required when using it.
Another example is an oral creatine formulation containing enteric-coated creatine hydrochloride and polyethylene glycol (Patent Document 2).
Patent Document 3 relates to a delivery system for a functional ingredient using a matrix, and describes a delivery system containing creatine, a coloring agent, a flavoring agent, and the like.

Thus, various studies have been conducted on methods for promoting creatine uptake, but there is a demand for products that allow creatine to be taken more easily, or that can be produced more simply and provide the full effects of creatine.

Medicine & Science in Sports & Exercise. 1998;30(7):1123-1129 Sports Nutrition: Answering "Why?" from the Basics of Science First Edition October 25, 2017 Nutrients. 2021;13(6):1915. Int J Sport Nutr Exerc Metab. 2003;13(3):294-302.

Special Publication No. 2007-536250 Patent No. 5433571 Special Publication No. 2005-527591 JP 2012-012409 A

The object of the present invention is to provide a composition that promotes creatine storage.

As a result of intensive research conducted by the inventors to solve the above problems, they discovered that the creatine storage effect can be further enhanced by combining a naturally occurring component that gives a yellow color to creatine compounds with isoleucine, and thus completed the present invention.

That is, the present invention provides:
(1) A composition comprising (a) a creatine compound, (b) a yellow-colored component derived from a natural product, and (c) isoleucine or a salt thereof;
(2) (a) The composition according to (1), wherein the creatine compound is creatine monohydrate;
(3) The composition according to (1), wherein the yellow-colored naturally derived component is at least one selected from the group consisting of crocin, lutein, safflower yellow, catechin, diosmin, zeaxanthin, and curcumin.
(4) The composition according to (1), which is for promoting creatine storage.
(5) The composition according to (1), which is for promoting energy storage.
(6) A creatine storage promoter comprising (b) a yellow-colored component derived from a natural product and (c) isoleucine or a salt thereof as active ingredients;
It is.

The present invention makes it possible to provide a composition that can promote creatine storage.

FIG. 1 is a graph showing that the test substance in Example 1 promotes creatine storage in a mouse myoblast cell line. The change rate of intracellular creatine amount corrected for protein amount is shown. Each group has 6 cases, and the average value of the change rate ± standard error is shown, with Comparative Example 1 taken as 100%. ***p<0.001 (Dunnett's multiple test using Comparative Example 1 as the control). 2 is a graph showing that the test substance promotes creatine storage in a mouse myoblast cell line in Example 2. The change rate of intracellular creatine amount corrected for protein amount is shown. Each group has 3 cases, and the average value of the change rate ± standard error is shown, with Comparative Example 1 taken as 100%. *p<0.05 (Student's t-test) 3 is a graph showing that the test substance promotes creatine storage in a mouse myoblast cell line in Example 3. The change rate of intracellular creatine amount corrected for protein amount is shown. Each group has 3 cases, and the average value of the change rate ± standard error is shown, with Comparative Example 1 taken as 100%. *p<0.05 (Student's t-test) 4 is a graph showing that the test substance promotes creatine storage in a mouse myoblast cell line in Example 4. The change rate of intracellular creatine amount corrected for protein amount is shown. Each group has 6 cases, and the average value of the change rate ± standard error is shown, with Comparative Example 1 taken as 100%. ***p<0.001 (Dunnett's multiple test using Comparative Example 1 as a control). 5 is a graph showing that the test substance promotes creatine storage in a mouse myoblast cell line in Example 5. The change rate of intracellular creatine amount corrected by protein amount is shown. Each group has 3 cases, and the average value of the change rate ± standard error is shown, with Comparative Example 1 taken as 100%. *p<0.05 (Student's t-test) 6 is a graph showing that the test substance promotes creatine storage in a mouse myoblast cell line in Example 6. The change rate of intracellular creatine amount corrected for protein amount is shown. Each group has 3 cases, and the average value of the change rate ± standard error is shown, with Comparative Example 1 taken as 100%. ***p<0.001 (Student's t-test) 7 is a graph showing that the test substance promotes creatine storage in a mouse myoblast cell line in Example 7. The change rate of intracellular creatine amount corrected for protein amount is shown. Each group has 3 cases, and the average value of the change rate ± standard error is shown, with Comparative Example 1 taken as 100%. *p<0.05 (Student's t-test) 8 is a graph showing that the test substances in Comparative Examples 5 and 6 do not change creatine storage in mouse myoblast cell lines. The change rate of intracellular creatine amount corrected by protein amount is shown. Each group has 3 cases, and the average value ± standard error of the change rate is shown with Comparative Example 1 set as 100%. n.s. No significant difference (Dunnett's multiple test with Comparative Example 1 as the control).

The component (a) creatine compound in the present invention refers to creatine or its derivatives, their salts, or their hydrates.The derivatives of creatine refer to esterified creatine, creatine with amino acids added, creatine with gluconic acid added, creatine with sugars added, cyclized creatine, etc.Preferably, creatine, creatine monohydrate, creatine anhydride, creatine phosphate, creatine citrate, creatine hydrochloride, creatine nitrate, creatine malate, creatine orotate, creatine pyruvate, creatine maleate, creatine phosphate, creatine nitrate, creatine maleate, creatine methyl ester, creatine ethyl ester, creatine sulfate, creatine leucinate, creatine gluconate, cyclocreatine, and polyethylene glycosylate creatine, more preferably, creatine, creatine monohydrate, creatine hydrochloride, creatine ethyl ester, creatine nitrate, creatine malate, and even more preferably, creatine monohydrate from the viewpoint of stability, etc.
These creatine compounds are commercially available as food or chemically synthesized products, and can be, for example, those manufactured by Alzchem, Tokyo Chemical Industry Co., Ltd., Fujifilm Wako Pure Chemical Industries, Ltd., etc. In addition, they may be synthesized from arginine, glycine, methionine, guanidinoacetic acid, sarcosinic acid, cyanamide, etc., or may be extracted from natural products containing creatine compounds.

The content of the creatine compound (a) in the composition of the present invention is not particularly limited, but is preferably 25 to 95 w/w%, more preferably 30 to 90 w/w%, and even more preferably 35 to 70 w/w%.

In the present invention, the naturally occurring components that exhibit a yellow color include crocin, lutein, β-carotene, safflower yellow, diosmin, isoflavone, catechin, curcumin, and zeaxanthin. The naturally occurring components that exhibit a yellow color have a maximum absorption wavelength of 250 to 490 nm.
There are no particular limitations on the safflower yellow, so long as it is a safflower-derived component that is normally used in foods, quasi-drugs, medicines, etc., and it may be one extracted from safflower, an annual plant of the Asteraceae family. Other names for safflower yellow include safflor yellow, safflower yellow pigment, and safflower pigment.
Crocin is a water-soluble carotenoid contained in gardenia, saffron, etc., and lutein is a type of fat-soluble carotenoid contained in green-yellow vegetables such as spinach and kale, and marigolds. β-carotene is a type of carotenoid with high fat solubility, and is contained in vegetables such as carrots and spinach. Diosmin is a type of flavonoid contained in rosemary, and the present invention also includes an embodiment in which rosemary extract is used as a pigment, and the rosemary pigment is not particularly limited as long as it is a rosemary-derived component that is normally used in foods, quasi-drugs, medicines, etc. Isoflavone is a type of flavonoid and is contained in large amounts in plants of the legume family. Catechin is a type of flavonoid mainly contained in tea. The catechin in the present invention is not particularly limited, but examples include epigallocatechin, epicatechin gallate, epicatechin, etc., and is preferably epigallocatechin gallate (EGCG). Curcumin is classified as a curcuminoid and is contained in turmeric, etc. Zeaxanthin, like lutein, is a type of carotenoid and is contained in leafy vegetables, corn, etc. The present invention also includes embodiments that use crushed products, squeezed juices, extracts, etc. of vegetables that contain these yellow-colored naturally derived components.

The content of the yellow-colored naturally derived component (b) in the composition of the present invention is not particularly limited, but is preferably 0.1 to 10 w/w%, more preferably 0.2 to 9 w/w%, and even more preferably 0.3 to 8 w/w%.

The component (c) isoleucine or a salt thereof in the present invention is not particularly limited as long as it is normally used in foods, quasi-drugs, pharmaceuticals, etc., and examples thereof include L-isoleucine, L-isoleucine ethyl ester hydrochloride, and L-isoleucine amide hydrochloride, and is preferably L-isoleucine.

The content of (c) isoleucine or a salt thereof in the composition of the present invention is not particularly limited, but is preferably 1 to 40 w/w%, more preferably 2 to 35 w/w%, and even more preferably 3 to 30 w/w%.

In the present invention, the mass ratio [(A):(B)] of the content of the creatine compound (A) to the content of the yellow naturally derived component (B) is preferably 1:0.001-1, more preferably 1:0.01-0.5, and even more preferably 1:0.01-0.2, from the viewpoint of promoting creatine storage in tissues. (b) Among the yellow naturally derived components, curcumin can obtain a creatine storage effect even in small amounts compared to other components, so the mass ratio [(A):(B)] is preferably 1:0.001-1, more preferably 1:0.003-0.5, and even more preferably 1:0.005-0.1.

In the present invention, the mass ratio of the content of the creatine compound (A) to the content of isoleucine or a salt thereof (C) [(A):(C)] is preferably 1:0.01-100, more preferably 1:0.05-50, and even more preferably 1:0.05-1, from the viewpoint of promoting creatine storage in tissues.

In the present invention, the mass ratio [(B):(C)] of the content of the yellow naturally derived component (B) to the content of isoleucine or a salt thereof (C) is preferably 1:0.01-100, more preferably 1:0.1-50, even more preferably 1:0.2-35, and even more preferably 1:0.2-10, from the viewpoint of promoting creatine storage in tissues. (b) Among the yellow naturally derived components, curcumin can obtain a creatine storage effect even in small amounts compared to other components, so the mass ratio [(B):(C)] is preferably 1:10-100, more preferably 1:20-90, and even more preferably 1:40-80.

The dosage of the composition of the present invention is not particularly limited, but the daily creatine intake for an adult is preferably 1.5 to 20 mg, more preferably 1.5 to 5 mg, and even more preferably 3 to 5 mg.

The composition of the present invention may contain vitamins, amino acids, minerals, herbal medicines and their extracts, etc., within the scope of not impairing the effects of the present invention. In addition, excipients, disintegrants, binders, lubricants, flow agents, antioxidants, coating agents, colorants, flavoring agents, etc. may be added, and further pH adjusters, cooling agents, suspending agents, antifoaming agents, thickening agents, solubilizing agents, surfactants, fragrances, etc. may be added as necessary.

The composition of the present invention can be used orally or parenterally.
The composition of the present invention can be used for foods, quasi-drugs, medicines, cosmetics, etc., and is preferably used as a food product.

When the composition of the present invention is used as an oral composition, its form may be solid or liquid, and is not particularly limited. For example, the composition may be provided as a powder, a powder, a granule, a tablet, a capsule, a dry syrup, a drink, a jelly, a beverage, a concentrated beverage, a solid effervescent beverage, a powdered beverage, or a jelly-like beverage.
When the composition of the present invention is used as a parenteral composition, it can be provided as an external preparation such as a lotion, cream, ointment, or patch, or as an injection.

The oral composition of the present invention can be prepared by a conventional method, and the method is not particularly limited.For example, in the case of a solid preparation, it can be prepared by a process of taking each component and mixing them uniformly.For example, in the case of a liquid preparation, it can be prepared by a process of taking each component, dissolving it in an appropriate amount of purified water, adjusting the pH appropriately, adding the remaining purified water to adjust the volume, filtering and sterilizing as necessary, and filling it into a container.
The parenteral composition of the present invention can also be prepared by a conventional method, and the method is not particularly limited.

The composition of the present invention has a creatine storage promoting effect that promotes creatine storage in tissues, and an energy storage promoting effect that promotes the storage of energy required for physical activities such as exercise.

The composition of the present invention is expected to promote creatine storage in tissues, thereby promoting energy production in tissues, improving explosive exercise performance, improving endurance exercise performance, improving muscle strength, improving muscle endurance, increasing muscle mass, improving basal metabolism, improving endurance to high-stress training, reducing fatigue during exercise, and improving recovery from fatigue. In addition, by reducing muscle atrophy or maintaining muscle mass, it is expected to maintain or improve symptoms associated with reduced muscle strength, reduced energy metabolism, and the like, such as sarcopenia and locomotive syndrome.

The composition of the present invention and its instructions may be labeled to the effect that the composition is used to increase the amount of creatine in tissues, promote creatine storage, and promote energy storage. Here, "labeled" includes being labeled on the body, container, packaging, etc. of a product containing the composition of the present invention, or being labeled in documents such as instructions disclosing product information, package inserts, pamphlets, posters, in-store POP, or other printed matter, and being labeled in advertisements used for promotion, including various leaflets, Internet, and television commercials. In addition, even if the composition is labeled to have other functions, or there is no indication of a function, the scope of the present invention includes those that substantially have the effect of promoting creatine storage in tissues or energy storage.

The creatine storage promoter of the present invention contains (b) a yellow-colored naturally derived component and (c) isoleucine or a salt thereof as active ingredients. The type, content, mass ratio [(B):(C)], etc. of (b) the yellow-colored naturally derived component and (c) isoleucine or a salt thereof are the same as those of the composition of the present invention. By ingesting the creatine storage promoter of the present invention together with food, drink, etc. containing a creatine compound, it is possible to promote the uptake and storage of creatine in tissues.

The present invention will be described in more detail below with reference to examples, but the present invention is not limited to these examples. In the following examples and comparative examples, creatine, safflower yellow, crocin, blue No. 1, phycocyanin, diosmin, and curcumin were manufactured by Tokyo Chemical Industry Co., Ltd., isoleucine and EGCG were manufactured by Fujifilm Wako Pure Chemical Industries, Ltd., lutein was manufactured by AstaTech Inc., and zeaxanthin was manufactured by AK Scientific, Inc.

The present invention will be described in more detail below with reference to examples, but the present invention is not limited to these examples.

Preparation of test samples (Examples 1 to 7, Comparative Examples 1 to 6)
Creatine, safflower yellow, crocin, lutein, EGCG, zeaxanthin, diosmin, curcumin, blue No. 1, phycocyanin, and isoleucine were weighed out to obtain the respective formulations shown in Table 1 below, and added to amino acid-free DMEM (FUJIFILM Wako Pure Chemical Industries, Ltd.) containing 10% Fetal Bovine Serum (FBS) to obtain test samples. However, lutein in Example 3, zeaxanthin in Example 5, and curcumin in Example 7 were dissolved in DMSO and then dissolved in the medium. These samples were heated to 37° C. using a hot water bath before the start of the evaluation.

Test Example 1: Evaluation in mouse myoblast cell line C2C12 cells Mouse myoblast cell line C2C12 cells (DS Pharma Biomedical) were seeded in a 6-well plate at 25,000 cells/ ^cm2 using a growth medium, DMEM (Fujifilm Wako Pure Chemical Industries, Ltd.) containing 10% fetal bovine serum (FBS), and cultured in an incubator set at 37°C and 5% _CO2 .
After one day of culture, the medium was removed by suction and replaced with the medium of the test sample shown in Examples 1 to 7 and Comparative Examples 1 to 6. After 4 hours of culture, the medium was removed, the Assay Buffer attached to the Creatine Assay Kit (Abcam, ab65339) was added, and the cells were collected. The amount of intracellular creatine was measured from the cell lysate using the Creatine Assay Kit (Abcam) according to the attached protocol. The amount of intracellular creatine was corrected by the amount of protein measured using the Protein Assay BCA Kit (Nacalai Tesque). The results were shown as the rate of change in the amount of creatine, with Comparative Example 1 taken as 100%. The rate of change in the amount of creatine was calculated as follows.
Rate of change in creatine amount (%)=(intracellular creatine amount/protein amount)/(intracellular creatine amount in Comparative Example 1/protein amount in Comparative Example 1)×100

The results are shown in Figures 1 to 8. For Comparative Example 1, data was obtained for each test and compared with each Example (when comparing with Examples 3, 5, and 7, DMSO was added to eliminate the effect of DMSO). For Comparative Example 3, data was also obtained for each test. From Figure 1, it was confirmed that the amount of intracellular creatine increased in Example 1 compared to Comparative Example 1, but the amount of intracellular creatine increased in Comparative Examples 2 and 3 compared to Comparative Example 1 was not confirmed. From Figure 2, it was confirmed that the amount of intracellular creatine increased in Example 2 compared to Comparative Example 1. From Figure 3, it was confirmed that the amount of intracellular creatine increased in Example 3 compared to Comparative Example 1. From Figure 4, it was confirmed that the amount of intracellular creatine increased in Example 4 compared to Comparative Example 1, but the amount of intracellular creatine increased in Comparative Examples 3 and 4 compared to Comparative Example 1 was not confirmed. From Figure 5, it was confirmed that the amount of intracellular creatine increased in Example 5 compared to Comparative Example 1. From Figure 6, it was confirmed that the amount of intracellular creatine increased in Example 6 compared to Comparative Example 1. From Figure 7, it was confirmed that the amount of intracellular creatine increased in Example 7 compared to Comparative Example 1. From FIG. 8, no increase in intracellular creatine was confirmed in Comparative Examples 5 and 6 compared to Comparative Example 1. Note that Blue No. 1 and phycocyanin used in Comparative Examples 5 and 6 are blue pigments used in foods, etc. It is presumed that the composition of the present invention has the effect of increasing the amount of creatine stored in muscle tissue.

<Prescription Examples>
Table 2 below lists examples of formulations for oral compositions of the present invention. The formulations which can be produced by the present invention increase creatine stores in the body when taken orally.

The components of the present invention that promote creatine storage in tissues can be used, for example, to improve explosive exercise performance, endurance exercise performance, etc., and to maintain or improve muscle strength, muscle endurance, muscle mass, basal metabolism, endurance to high-load training, and ability to recover from fatigue.
In addition, since creatine storage promoters have the effect of promoting energy storage in bodily tissues such as skeletal muscles, they are also effective in maintaining or improving cognitive function in the brain and preventing or improving wrinkles in skin tissue (see Patent Document 4).
Furthermore, since sufficient energy can be stored in tissues even if the amount of creatine ingested is reduced, gastrointestinal symptoms caused by ingesting a large amount of creatine can be alleviated.

Claims (6)

A composition containing (a) a creatine compound, (b) a yellow-colored component derived from a natural product, and (c) isoleucine or a salt thereof. (a) The composition according to claim 1, wherein the creatine compound is creatine monohydrate. (b) The composition according to claim 1 or 2, wherein the naturally occurring component that exhibits a yellow color is at least one selected from the group consisting of crocin, lutein, safflower yellow, catechin, diosmin, zeaxanthin, and curcumin. The composition according to claim 1, which is for promoting creatine storage. The composition according to claim 1 for promoting energy storage. A creatine storage promoter comprising (b) a yellow-colored naturally-derived component and (c) isoleucine or a salt thereof as active ingredients.

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