Composition for improving, maintaining, and / or preventing deterioration of liver function

Abstract

This composition for improving, maintaining, and / or preventing the deterioration of liver function comprises a tetrapeptide consisting of the amino acid sequence represented by SEQ ID NO: 1, and is for healthy humans.

Description

Composition for improving, maintaining, and / or preventing deterioration of liver function

[0001] The present invention relates to a composition for improving, maintaining, and / or preventing deterioration of liver function.

[0002] The liver is an important organ in the human body for maintaining homeostasis. Decrease in liver function not only leads to deterioration of nutritional status accompanied by decreases in glucose metabolism, lipid metabolism, protein synthesis ability, etc., but also affects important biological functions such as detoxification and excretion. It is known that continuous high values of aspartate aminotransferase (AST) or alanine aminotransferase (ALT) used as indicators of liver function are associated with chronic liver diseases (such as liver cirrhosis, liver cancer, etc.).

[0003] Since globin proteolysate suppresses the increase in postprandial blood neutral fat levels, it is used in foods for specified health uses, foods with functional claims, etc.

[0004] Regarding globin proteolysate, Patent Document 1 discloses that globin proteolysate had an effect of improving liver damage on a mouse alcoholic liver injury model and a mouse acute liver injury model. However, it is not known whether globin proteolysate has an effect of improving liver function on humans.

[0005] Japanese Patent Application Laid-Open No. 2006-287569

[0006] An object of the present invention is to provide a composition capable of improving, maintaining, and / or preventing deterioration of liver function. Another object of the present invention is to provide a composition capable of improving, maintaining, and / or preventing deterioration of blood alanine aminotransferase (ALT) level or blood aspartate aminotransferase (AST) level.

[0007] The present inventors have found that a composition containing a tetrapeptide consisting of the amino acid sequence (tryptophan-threonine-glutamine-arginine) represented by Sequence ID No. 1 (for example, a globin protein hydrolysate) can improve, maintain, and / or prevent deterioration of liver function in healthy humans. Furthermore, the present inventors have found that the above composition can improve, maintain, and / or prevent deterioration of liver function marker values ​​(serum ALT and serum AST).

[0008] This disclosure provides, for example, the inventions described in [1] to

[26] below: [1] A composition for improving, maintaining and / or preventing deterioration of liver function, for use in healthy humans, comprising a tetrapeptide consisting of the amino acid sequence represented by Sequence ID No. 1. [2] A composition for improving, maintaining and / or preventing deterioration of blood alanine aminotransferase (ALT) or aspartate aminotransferase (AST) levels, for use in healthy humans, comprising a tetrapeptide consisting of the amino acid sequence represented by Sequence ID No. 1. [3] The composition according to [1] or [2], wherein the composition contains a globin protein hydrolysate, and at least a portion of the tetrapeptide is derived from the globin protein hydrolysate. [4] The composition according to any one of [1] to [3], wherein the human is a person whose blood triglyceride level is 120 mg / dL or more and less than 200 mg / dL. [5] The composition according to any one of [1] to [4], wherein the human is a person who drinks alcohol. [6] The composition according to any one of [1] to [5], wherein the person is a person whose blood ALT level is 20 U / L or more and 50 U / L or less. [7] The composition according to any one of [1] to [6], wherein the person is a person whose blood AST level is 50 U / L or less. [8] The composition according to any one of [1] to [7], which is used to be ingested or administered in an amount of 5 mg or more and 15 mg or less per day in terms of the tetrapeptide. [9] The composition according to any one of [1] to [8], which is used to be ingested or administered for 4 weeks or more.

[10] A method for improving, maintaining and / or preventing deterioration of liver function in a subject that requires the ingestion or administration of a composition containing a tetrapeptide consisting of an amino acid sequence represented by Sequence ID No. 1, wherein the subject is a healthy person.

[11] A method for improving, maintaining and / or preventing deterioration of a subject's blood alanine aminotransferase level or blood aspartate aminotransferase level, comprising ingesting or administering a composition containing a tetrapeptide comprising the amino acid sequence represented by Sequence ID No. 1, wherein the subject is a healthy human being.

[12] The method according to

[10] or

[11] , wherein the composition contains a globin protein hydrolysate.

[13] A tetrapeptide consisting of the amino acid sequence represented by Sequence ID No. 1 for use in improving, maintaining, and / or preventing deterioration of the liver function of the target. [13a] A composition containing a tetrapeptide consisting of the amino acid sequence represented by Sequence ID No. 1 for use in improving, maintaining, and / or preventing deterioration of the liver function of the target.

[14] A tetrapeptide consisting of the amino acid sequence represented by Sequence ID No. 1 for use in improving, maintaining, and / or preventing deterioration of the blood alanine aminotransferase level or blood aspartate aminotransferase level of the target. [14a] A composition containing a tetrapeptide consisting of the amino acid sequence represented by Sequence ID No. 1 for use in improving, maintaining, and / or preventing deterioration of the blood alanine aminotransferase level or blood aspartate aminotransferase level of the target.

[15] A globin protein hydrolysate containing the above tetrapeptide for use in improving, maintaining, and / or preventing deterioration of the liver function of the target. [15a] A composition containing a globin protein hydrolysate containing the above tetrapeptide for use in improving, maintaining, and / or preventing deterioration of the liver function of the target.

[16] A globin protein hydrolysate containing the above tetrapeptide for use in improving, maintaining, and / or preventing deterioration of target blood alanine aminotransferase levels or blood aspartate aminotransferase levels. [16a] A composition containing a globin protein hydrolysate containing the above tetrapeptide for use in improving, maintaining, and / or preventing deterioration of target blood alanine aminotransferase levels or blood aspartate aminotransferase levels.

[17] Use of a tetrapeptide consisting of the amino acid sequence represented by Sequence ID No. 1 for the production of a composition for healthy humans to improve, maintain, and / or prevent deterioration of liver function.

[18] Use of a tetrapeptide consisting of the amino acid sequence represented by Sequence ID No. 1 for the production of a composition for healthy humans to improve, maintain, and / or prevent deterioration of blood alanine aminotransferase levels or blood aspartate aminotransferase levels.

[19] Use of a globin protein hydrolysate for the production of a composition for healthy humans to improve, maintain, and / or prevent deterioration of liver function.

[20] Use of globin proteolytic product for the manufacture of a composition for healthy human beings for improving, maintaining and / or preventing deterioration of blood alanine aminotransferase levels or blood aspartate aminotransferase levels.

[21] The human being is a person whose blood triglyceride level is 120 mg / dL or more and less than 200 mg / dL, and the method according to any of

[11] to

[20] , tetrapeptide, globin proteolytic product, or use.

[22] The human being is a person who drinks alcohol, and the method according to any of

[11] to

[21] , tetrapeptide, globin proteolytic product, or use.

[23] The human being is a person whose blood ALT level is 20 U / L or more and 50 U / L or less, and the method according to any of

[11] to

[22] , tetrapeptide, globin proteolytic product, or use.

[24] The human being is a person whose blood AST level is 50 U / L or less, and the method according to any of

[11] to

[23] , tetrapeptide, globin proteolytic product, or use.

[25] The tetrapeptide or globin protein hydrolysate is used in such a manner that it is taken in or administered at a daily dose of 5 mg or more and 15 mg or less in terms of the tetrapeptide, according to any of the methods described in

[11] to

[24] , the tetrapeptide, the globin protein hydrolysate, or the use thereof.

[26] The tetrapeptide or globin protein hydrolysate is used in such a manner that it is taken in or administered for four weeks or more, according to any of the methods described in

[11] to

[25] , the tetrapeptide, the globin protein hydrolysate, or the use thereof.

[0009] The present invention provides a composition that can improve, maintain, and / or prevent deterioration of liver function in healthy individuals. Furthermore, the present invention provides a composition that can improve, maintain, and / or prevent deterioration of liver function marker values ​​(serum ALT or serum AST) in healthy individuals.

[0010] The embodiments of this disclosure will be described in detail below.

[0011] The composition for improving, maintaining, and / or preventing deterioration of liver function according to this embodiment, and the composition for improving, maintaining, and / or preventing deterioration of blood ALT or blood AST levels according to this embodiment (hereinafter, "the composition for improving, maintaining, and / or preventing deterioration of liver function according to this embodiment" and "the composition for improving, maintaining, and / or preventing deterioration of blood ALT or blood AST levels according to this embodiment" will be collectively referred to simply as "the composition according to this embodiment") contain a tetrapeptide consisting of the amino acid sequence represented by Sequence ID No. 1. The composition according to this embodiment may be used in healthy humans.

[0012] The composition according to this embodiment can reduce abnormally high blood ALT or AST levels in subjects who ingest or are administered it, and therefore can improve, maintain, and / or prevent deterioration of blood ALT or AST levels.

[0013] Improving blood ALT or AST levels means reducing a person's blood ALT or AST levels that are high outside the normal range to bring them within the normal range or closer to the normal range, or reducing a person's blood ALT or AST levels that are slightly elevated within the normal range. Maintaining blood ALT or AST levels means suppressing an increase in a person's blood ALT or AST levels and maintaining them within the normal range. Preventing deterioration of blood ALT or AST levels means reducing blood ALT or AST levels that are high outside the normal range or slightly elevated within the normal range, thereby preventing those blood ALT or AST levels from rising further.

[0014] Serum ALT and serum AST levels are known liver function markers. Serum ALT and serum AST are enzymes produced by liver cells and are released into the bloodstream when liver cells are damaged. Therefore, improvement, maintenance, and / or prevention of deterioration of serum ALT or serum AST levels indicate improvement, maintenance, and / or prevention of deterioration of liver function. Conversely, high serum ALT or serum AST levels suggest liver damage.

[0015] In this disclosure, the term "serum ALT value" refers to the concentration (U / L) of ALT in the blood as measured by the following blood biochemistry test. In the above blood biochemistry test, the serum ALT value is measured according to the JSCC standardized method.

[0016] In this disclosure, "blood AST value" refers to the concentration (U / L) of AST in the blood as measured by the blood biochemistry test described below. In the blood biochemistry test described above, the blood AST value is measured according to the JSCC standardized method.

[0017] The "Standard Health Checkup and Health Guidance Program (FY2024 Edition)" (Ministry of Health, Labour and Welfare) designates a blood ALT level of 31 U / L as the threshold for health guidance and a blood ALT level of 51 U / L as the threshold for recommending a medical consultation (https: / / www.mhlw.go.jp / content / 12401000 / 000967510.pdf, accessed July 19, 2024). Furthermore, according to the Japan Society for Human Dock and Preventive Medicine (https: / / www.ningen-dock.jp / public_method / , accessed August 6, 2024), a blood ALT level of 30 U / L or less is considered the normal range, and a blood ALT level between 31 U / L and 50 U / L is considered the range requiring attention. Based on these, in this disclosure, the normal range for human blood ALT levels (healthy range for human blood ALT levels) is defined as 50 U / L or less.

[0018] The "Standard Health Checkup and Health Guidance Program (FY2024 Edition)" (Ministry of Health, Labour and Welfare) defines a blood AST value of 31 U / L as the threshold for health guidance and a blood AST value of 51 U / L as the threshold for recommending a medical examination (https: / / www.mhlw.go.jp / content / 12401000 / 000967510.pdf, accessed July 19, 2024). Furthermore, according to the Japan Society for Human Dock and Preventive Medicine (https: / / www.ningen-dock.jp / public_method / , accessed August 6, 2024), a blood AST value of 30 U / L or less is considered the normal range. Based on these, in this disclosure, the normal range of human blood AST values ​​(healthy range of human blood AST values) is defined as 50 U / L or less.

[0019] Whether the composition according to this embodiment can improve, maintain, and / or prevent deterioration of liver function can be determined based on whether it can improve blood ALT levels or blood AST levels. Whether the above composition can improve blood ALT levels or blood AST levels can be determined, for example, by administering the above composition to 35 to 40 healthy individuals with blood ALT levels of 20 to 50 U / L in an appropriate dosage and manner, and whether the blood ALT levels or blood AST levels of the above individuals can be reduced. For example, if the rate of change in blood ALT value due to ingestion of the above composition (the average value of the percentage of the change in blood ALT value due to ingestion of the above composition, calculated for each person, relative to the blood ALT value before ingestion) is -0.1% or less, -0.2% or less, -0.3% or less, -0.5% or less, -0.7% or less, -1% or less, -1.2% or less, -1.4% or less, -2.5% or less, or -10% or less, or if the rate of change in blood AST value due to ingestion of the composition ( If the average percentage (%) of the change in blood AST value due to ingestion of the above composition, calculated for each individual, relative to the blood AST value before ingestion, is -0.1% or less, -0.2% or less, -0.3% or less, -0.5% or less, -0.7% or less, -1% or less, -1.2% or less, -1.4% or less, -2.5% or less, or -10% or less, then it can be determined that the above composition can improve the blood ALT value or blood AST value of a healthy person.

[0020] In this disclosure, "healthy person" may mean, for example, a person who has no history of suffering from a serious liver disease (viral hepatitis, drug-induced liver injury, cirrhosis, etc.), a person who does not suffer from the above-mentioned serious liver disease, or a person who meets all of these conditions.

[0021] The composition according to this embodiment may be used for healthy humans. The humans may be, for example, 18 years of age or older, 20 years of age or older, 40 years of age or older, 20 years of age or older but under 65 years of age, or 40 years of age or older but under 65 years of age. The humans may be female or male. Furthermore, the humans may be Asian or Japanese.

[0022] The above-mentioned person is preferably a person whose blood triglyceride level is 120 mg / dL or more and less than 200 mg / dL, or a person whose blood triglyceride level is 120 mg / dL or more and 199 mg / dL or less. As shown in the examples described below, when a person with blood triglyceride levels in this range ingests the composition according to this embodiment, the improvement effect on blood ALT and blood AST levels is significant. In this disclosure, "blood triglyceride level" means the concentration of triglycerides in the blood (mg / dL) measured by the blood biochemistry test described below. The above blood biochemistry test is performed by an enzymatic method.

[0023] According to Attachment 2 of the response regarding the partial amendment to "Approval of Labeling for Foods for Specified Health Uses," a blood triglyceride level of 120 mg / dL to 149 mg / dL is considered "high normal range," and a blood triglyceride level of 150 mg / dL to 199 mg / dL is considered "slightly high." Therefore, blood triglyceride levels of 120 mg / dL to less than 200 mg / dL and 120 mg / dL to 199 mg / dL can be considered high blood triglyceride levels within the normal range.

[0024] The person described above is preferably a person who drinks alcohol. As shown in the examples described later, when a person who drinks alcohol ingests the composition according to this embodiment, the improvement effect on blood ALT and blood AST values ​​is significant. A person who drinks alcohol may be a person who drinks alcohol during the period in which the above composition is administered or ingested, or a person who has a drinking habit. The former person who drinks alcohol may be a person whose average daily intake of pure alcohol during the above period is 0.4 g or more and 42.2 g or less. The latter person who drinks alcohol may have a drinking habit in which their average daily intake of pure alcohol is 0.4 g or more and 42.2 g or less. The amount of pure alcohol ingested is the product of the alcohol content (volume %) of the alcohol consumed, multiplied by the volume of the alcohol and the specific gravity of alcohol (0.8).

[0025] The above-mentioned individuals may be those with elevated blood ALT levels outside the normal range, those with levels within the normal range, those with levels between 20 U / L and 50 U / L, or those with levels between 25 U / L and 49 U / L. A blood ALT level between 20 U / L and 50 U / L can be considered a slightly elevated ALT level within the normal range. Therefore, if the blood ALT levels of individuals in this range can be reduced, it is expected that future liver disease can be prevented.

[0026] The healthy individuals mentioned above may have blood AST levels that are high but outside the normal range, or within the normal range, or below 50 U / L, or between 18 U / L and 51 U / L. Reducing slightly elevated blood AST levels within the normal range is expected to help prevent future liver disease.

[0027] In the examples described later, it has been demonstrated that globin proteolytic products can improve human blood ALT or blood AST levels. Sasagawa et al. (Yuka Sasagawa. "Hepatoprotective effects of globin proteolytic products and their active ingredient Trp-Thr-Gln-Arg on a galactosamine-induced liver injury model in SD rats." Pharmacology and Therapeutics 39.8 (2011): 741-749.) have disclosed that in non-clinical studies using SD rats, a tetrapeptide consisting of the amino acid sequence represented by SEQ ID NO: 1 exerted hepatoprotective effects by improving the antioxidant defense mechanism and inflammatory state of the liver in SD rats. Previously, it was unclear whether the above-mentioned tetrapeptide could improve human blood ALT or AST levels. However, considering that globin protein hydrolysates were able to improve human blood ALT or AST levels, that the above-mentioned globin protein hydrolysates contain the above-mentioned tetrapeptide, and that the above-mentioned tetrapeptide can exert a hepatoprotective effect in rats, it can be said that the improvement of human blood ALT or AST levels by globin protein hydrolysates is due to the hepatoprotective effect of the above-mentioned tetrapeptide.

[0028] The composition according to this embodiment may contain globin proteolytic product. Since globin proteolytic product contains a tetrapeptide consisting of the amino acid sequence represented by Sequence ID No. 1, if the composition contains globin proteolytic product, at least a portion of the tetrapeptide contained in the composition will be derived from the globin proteolytic product. Alternatively, if the composition contains globin proteolytic product, all of the tetrapeptide contained in the composition may be derived from the globin proteolytic product.

[0029] Globin protein hydrolysates are the breakdown products of globin proteins such as hemoglobin and myoglobin. The above-mentioned hydrolysates may also be enzymatic hydrolysates.

[0030] The globin protein hydrolysate contains a tetrapeptide consisting of the amino acid sequence represented by Sequence ID No. 1, but the content of the above tetrapeptide in the globin protein hydrolysate may be 0.1% by mass or more and 5% by mass or less, or 0.3% by mass or more and 2% by mass or less.

[0031] The globin protein hydrolysate may contain not only the tetrapeptide consisting of the amino acid sequence represented by SEQ ID NO: 1, but also other tetrapeptides such as valine-valine-tyrosine-proline (SEQ ID NO: 2).

[0032] Globin proteins are not particularly limited, but may be derived from the blood or meat (livestock meat, fish meat) of animals such as mammals (cattle, pigs, sheep, humans, horses, etc.) and fish. Blood typically contains a large amount of hemoglobin, while meat typically contains a large amount of myoglobin.

[0033] The method for producing globin protein hydrolysates is not particularly limited, but for example, it may be a method in which a globin protein-containing substance such as blood or meat is added to water to form a dispersion, and a hydrolytic enzyme is added to the dispersion to allow the hydrolysis reaction of the globin protein to proceed.

[0034] The amount of the globin protein-containing substance in the above dispersion may be, for example, 5% by mass or more and 30% by mass or less, based on the solid content (components other than water) of the globin protein-containing substance.

[0035] As the hydrolytic enzymes mentioned above, acidic proteases, neutral proteases, alkaline proteases, or combinations thereof may be used.

[0036] The above hydrolysis reaction may be carried out at the optimal pH of the hydrolytic enzyme used. The above hydrolysis reaction may be carried out at the optimal temperature of the hydrolytic enzyme used (for example, 20°C to 70°C). The above hydrolysis reaction may be carried out for, for example, 3 hours to 48 hours. There are no particular restrictions on the method of adding the hydrolytic enzyme to the above dispersion, but the hydrolytic enzyme may be added all at once or sequentially.

[0037] The above hydrolysis reaction yields a globin protein hydrolysate dispersed in water. This globin protein hydrolysate may be used as is, dried, or dried and solidified with an appropriate amount of a bulking agent such as carboxymethylcellulose or dextrin. Furthermore, the globin protein hydrolysate may be purified as needed.

[0038] Furthermore, globin protein hydrolysates may be produced by the method described in International Publication No. 89 / 06970.

[0039] The composition according to this embodiment may be used for oral ingestion or administration. That is, the composition may be an oral preparation.

[0040] In the composition according to this embodiment, the content of the tetrapeptide consisting of the amino acid sequence represented by Sequence ID No. 1 may be, for example, 0.01% by mass or more and 99% by mass or less, or 0.1% by mass or more and 90% by mass or less.

[0041] If the composition according to this embodiment is a tablet, the content of the tetrapeptide consisting of the amino acid sequence represented by Sequence ID No. 1 in the tablet may be, for example, 0.05 w / w% or more, 0.01 w / w% or more, 0.1 w / w% or more, 0.25 w / w% or more, or 0.3 w / w% or more, and may also be 6 w / w% or less, 5 w / w% or less, 2 w / w% or less, 1 w / w% or less, or 0.5 w / w% or less.

[0042] When the composition according to this embodiment is a liquid preparation, the content of the tetrapeptide consisting of the amino acid sequence represented by SEQ ID NO: 1 in the liquid preparation may be, for example, 0.00005 w / v% or more, 0.0001 w / v% or more, 0.0005 w / v% or more, 0.001 w / v% or more, 0.0015 w / v% or more, or 0.002 w / v% or more, and may also be 1 w / v% or less, 0.5 w / v% or less, 0.1 w / v% or less, 0.05 w / v% or less, or 0.02 w / v% or less.

[0043] When the composition according to this embodiment is a granule preparation, the content of the tetrapeptide consisting of the amino acid sequence represented by SEQ ID NO: 1 in the granule preparation may be, for example, 0.05 w / w%, 0.1 w / w% or more, 0.2 w / w% or more, or 0.25 w / w% or more, and may also be 10 w / w% or less, 5 w / w% or less, 1 w / w% or less, or 0.5 w / w% or less.

[0044] When the composition according to this embodiment contains a globin proteolysate, the content of the globin proteolysate in the composition may be, for example, 1 mass% or more and 99 mass% or less, and may also be 10 mass% or more and 90 mass% or less.

[0045] When the composition according to this embodiment is a tablet, the content of the globin proteolysate in the tablet may be, for example, 1 w / w% or more, 2 w / w% or more, 5 w / w% or more, 10 w / w% or more, 20 w / w% or more, or 50 w / w% or more, and may also be 99 w / w% or less, 96 w / w% or less, 91 w / w% or less, 89 w / w% or less, 70 w / w% or less, 40 w / w% or less, or 20 w / w% or less.

[0046] When the composition according to this embodiment is a liquid preparation, the content of the globin proteolysate in the liquid preparation may be, for example, 0.01 w / v% or more, 0.02 w / v% or more, 0.1 w / v% or more, 0.2 w / v% or more, 0.3 w / v% or more, or 0.4 w / v% or more, and may also be 25 w / v% or less, 20 w / v% or less, 10 w / v% or less, 8 w / v% or less, or 5 w / v% or less.

[0047] When the composition according to this embodiment is a granule, the content of the above globin proteolysate in the granule may be, for example, 10 w / w% or more, 15 w / w% or more, 20 w / w% or more, 30 w / w% or more, 40 w / w% or more, or 50 w / w% or more, and may be 99 w / w% or less, 97 w / w% or less, 91 w / w% or less, 85 w / w% or less, or 80 w / w% or less.

[0048] The composition according to this embodiment may be used such that, in terms of the tetrapeptide consisting of the amino acid sequence represented by SEQ ID NO: 1, for example, 1 mg or more and 30 mg or less is ingested or administered per day, 3 mg or more and 20 mg or less is ingested or administered per day, 5 mg or more and 15 mg or less, or 10 mg is ingested or administered per day.

[0049] When the composition according to this embodiment contains a globin proteolysate, the above composition may be used such that, in terms of the globin proteolysate, for example, 0.5 g or more and 4 g or less is ingested or administered per day, 1 g or more and 3 g or less is ingested or administered per day, 1.5 g or more and 2.5 g or less, or 2 g is ingested or administered per day.

[0050] The composition according to this embodiment may be used such that it is ingested or administered for 4 weeks or more, or 8 weeks or more. When the above composition is ingested or administered to a healthy human whose blood neutral fat value is 120 mg / dL or more and less than 200 mg / dL, or a human who drinks alcohol, by ingesting or administering the above composition every day for 8 weeks or more, the blood ALT value and the blood AST value are significantly improved. The above ingestion or administration dose and the above ingestion or administration period can be arbitrarily combined.

[0051] The composition according to this embodiment may further contain pharmacologically active ingredients or physiologically active ingredients, as long as the effects of the present invention are not impaired. Specific examples of such pharmacologically active ingredients or physiologically active ingredients include, for example, ubiquinone (coenzyme Q10), vitamin B6, vitamin B12, vitamin C, vitamin D, vitamin A, vitamin E, niacin, calcium pantothenate, taurine, carnosine, anserine, balenine, citrulline, gamma-aminobutyric acid, valine, leucine, isoleucine, glycine, arginine, ornithine, glutamic acid, glutamine, creatine, carnitine, and ru Examples include theolin, quercetin, genistin, cyanidin, resveratrol, diosgenin, isoflavone aglycone, lipoic acid, zinc, iron, calcium, selenium, astaxanthin, zeaxanthin, beta-carotene, myristic acid, palmitic acid, stearic acid, oleic acid, linoleic acid, linolenic acid, arachidonic acid, placenta extract, pycnogenol, curcumin, bisacroline, sulforaphane, liver extract, liver hydrolysate, GABA, etc. From the viewpoint of further enhancing the effects of the present invention, preferred pharmacologically active or physiologically active ingredients include ubiquinone (coenzyme Q10), vitamin B1, vitamin B2, vitamin B6, vitamin B12, vitamin C, vitamin E, astaxanthin, zinc, carnitine, ornithine, curcumin, bisacroline, sulforaphane, liver extract, liver hydrolysate, or GABA, with ubiquinone (coenzyme Q10), astaxanthin, zinc, ornithine, curcumin, or GABA being more preferred. The pharmacologically active or physiologically active ingredients may be used individually or in combination of two or more.

[0052] The composition according to this embodiment may further contain various additives, as long as they do not impair the effects of the present invention. Specific examples of such additives include, for example, excipients, binders, disintegrants, lubricants, colorants, flavoring agents, odorants, sugars, sugar alcohols / polyhydric alcohols, high-intensity sweeteners, oils and fats, emulsifiers, thickeners, acidulants, fruit juices, etc. Examples of excipients include lactose, sucrose, sodium chloride, glucose, starch, gelatin, calcium carbonate, kaolin, crystalline cellulose, and silicic acid. Examples of binders include water, ethanol, propanol, simple syrup, glucose solution, starch solution, gelatin solution, cellulose, carboxymethylcellulose, hydroxypropylcellulose, hydroxypropylmethylcellulose, hydroxypropyl starch, methylcellulose, ethylcellulose, calcium phosphate, and polyvinylpyrrolidone. Examples of disintegrants include dried starch, sodium alginate, agar powder, sodium bicarbonate, calcium carbonate, sodium lauryl sulfate, monoglyceride stearate, and lactose. Examples of lubricants include talc, stearate, borax, and polyethylene glycol. Examples of flavoring agents include sucrose, orange peel, citric acid, and tartaric acid. Examples of sugars include sucrose, isomerized sugar, glucose, fructose, palatinose, trehalose, lactose, and xylose. Examples of sugar alcohols and polyhydric alcohols include sorbitol, xylitol, erythritol, lactitol, maltitol, palatinose, reduced starch syrup, reduced maltose syrup, glycerin, and propylene glycol. Examples of high-intensity sweeteners include aspartame, stevia, acesulfame potassium, and sucralose. Examples of oils and fats include safflower oil, grape seed oil, sunflower oil, olive oil, corn oil, sesame oil, soybean oil, rapeseed oil, perilla oil, and rice oil. Examples of emulsifiers include sucrose fatty acid esters, glycerin fatty acid esters, and lecithin. Examples of thickeners include carrageenan, xanthan gum, guar gum, pectin, and locust bean gum. Examples of acidulants include citric acid, lactic acid, and malic acid. Examples of fruit juices include lemon juice, orange juice, and berry juices.Examples of fragrances include citrus fragrances and black tea fragrances. Additives may be used individually or in combination of two or more.

[0053] The composition according to this embodiment can be used, for example, as an ingredient in food and beverages (drinks, food products), pharmaceuticals, quasi-drugs, etc. Furthermore, the composition according to this embodiment can also be used, for example, as a functional food, health supplement, food preparation (e.g., confectionery tablets), food for specified health uses, nutritional functional foods, special dietary foods, pharmaceutical preparations, quasi-drug preparations, etc.

[0054] Furthermore, when the composition according to this embodiment is used as food or beverage, the food may be a general food or beverage blended with a tetrapeptide consisting of the amino acid sequence represented by SEQ ID NO: 1 and other components as needed. Examples of such food and beverages include cookies, biscuits, snack foods, jellies, gummies, chocolates, gum, candies, cheese, noodles such as ramen, seasonings, processed meat products, frozen foods, instant foods, french fries, nutritional drinks, juices, tea beverages, coffee beverages, dairy beverages, and alcoholic beverages.

[0055] Furthermore, the composition according to this embodiment may be in any dosage form depending on the purpose. Examples of such dosage forms include oral preparations such as tablets (including orally disintegrating tablets, chewable tablets, lozenges, etc.), granules, powders, soft capsules, hard capsules, lozenges, jellies, or liquids (including suspensions, emulsions, syrups, etc.); external preparations such as ointments, suppositories, patches, sprays, etc.; and injectable preparations.

[0056] The composition according to this embodiment can improve blood ALT or blood AST levels and can therefore be used as a liver protective agent. Furthermore, the composition according to this embodiment may be labeled with statements such as, "Recommended for people who drink alcohol," "Recommended for people with elevated liver enzyme levels," "Recommended for people with elevated blood triglycerides," "Reduces slightly elevated liver enzyme (ALT) levels within the normal range," "Reduces slightly elevated liver enzyme (ALT and AST) levels within the normal range," "Reduces slightly elevated liver enzyme (ALT and AST) levels due to alcohol," "Reduces slightly elevated liver enzyme (ALT and AST) levels due to alcohol," or "Reduces slightly elevated liver enzyme (ALT and AST) levels due to drinking." Note that liver enzyme levels are synonymous with liver function marker levels and refer to blood ALT or blood AST levels. Also, the blood triglycerides refer to blood triglyceride levels.

[0057] The composition according to this embodiment is suitably used for people who want to take care of their liver, those who are concerned about their liver, those who are concerned about their liver function values, those who are concerned about drinking too much, those who are concerned about decreased liver function, and those who want to prevent hangovers. It is also suitably used in products that include illustrations of the liver or alcoholic beverages on the packaging, and in products that include the word "liver" in the product name, etc.

[0058] The present invention can also be understood as a method for improving, maintaining, and / or preventing deterioration of liver function in a subject requiring the use of a composition containing a tetrapeptide comprising the amino acid sequence represented by Sequence ID No. 1, wherein the subject is a healthy human being. The present invention can also be understood as a tetrapeptide comprising the amino acid sequence represented by Sequence ID No. 1 for use in improving, maintaining, and / or preventing deterioration of liver function in a subject. The present invention can also be understood as a composition containing a tetrapeptide comprising the amino acid sequence represented by Sequence ID No. 1 for use in improving, maintaining, and / or preventing deterioration of liver function in a subject. The present invention can also be understood as a globin protein hydrolysate containing the above tetrapeptide for use in improving, maintaining, and / or preventing deterioration of liver function in a subject. The present invention can also be understood as the use of a tetrapeptide comprising the amino acid sequence represented by Sequence ID No. 1 for the production of a composition for healthy humans, for improving, maintaining, and / or preventing deterioration of liver function. The present invention can also be understood as the use of globin proteolytics for the production of compositions for use in healthy humans and for improving, maintaining, and / or preventing deterioration of liver function. The methods, tetrapeptides, globin proteolytics, and specific embodiments of use can be applied without limitation to the embodiments described as specific embodiments of the compositions according to this embodiment.

[0059] The present invention can also be understood as a method for improving, maintaining, and / or preventing deterioration of a target's blood alanine aminotransferase level or blood aspartate aminotransferase level, comprising ingesting or administering a composition containing a tetrapeptide comprising the amino acid sequence represented by Sequence ID No. 1 to a target requiring such composition, wherein the target is a healthy human being. The present invention can also be understood as a tetrapeptide comprising the amino acid sequence represented by Sequence ID No. 1 for use in improving, maintaining, and / or preventing deterioration of a target's blood alanine aminotransferase level or blood aspartate aminotransferase level. The present invention can also be understood as a composition containing a tetrapeptide comprising the amino acid sequence represented by Sequence ID No. 1 for use in improving, maintaining, and / or preventing deterioration of a target's blood alanine aminotransferase level or blood aspartate aminotransferase level. The present invention can also be understood as a globin protein hydrolysate containing the above tetrapeptide for use in improving, maintaining, and / or preventing deterioration of a target's blood alanine aminotransferase level or blood aspartate aminotransferase level. The present invention can also be considered as a composition containing a globin protein hydrolysate containing the above-mentioned tetrapeptide for use in improving, maintaining, and / or preventing deterioration of target blood alanine aminotransferase levels or blood aspartate aminotransferase levels. The present invention can also be considered as the use of a tetrapeptide consisting of the amino acid sequence represented by SEQ ID NO: 1 for the production of a composition for use in healthy humans for improving, maintaining, and / or preventing deterioration of blood alanine aminotransferase levels or blood aspartate aminotransferase levels. The present invention can also be considered as the use of a globin protein hydrolysate for the production of a composition for use in healthy humans for improving, maintaining, and / or preventing deterioration of blood alanine aminotransferase levels or blood aspartate aminotransferase levels. The methods, tetrapeptides, globin protein hydrolysates, and specific embodiments of use can be applied without limitation to the embodiments described as specific embodiments of the composition according to this embodiment.

[0060] The present invention will be described in more detail below with reference to examples.

[0061] In this study, a randomized, placebo-controlled, double-blind, parallel-group comparative trial was conducted to evaluate the effects of ingesting a test food containing globin protein hydrolysate on blood ALT and blood AST levels.

[0062] This study was approved by the Fukuzumi Internal Medicine Clinic Institutional Review Board on August 7, 2023, and after obtaining written consent from the subjects, it was conducted in accordance with the spirit of the Declaration of Helsinki (revised October 2013), the Ethical Guidelines for Life Science and Medical Research Involving Human Subjects (partially revised March 27, 2023), and the study protocol for this study (Registration Number UMIN000051963).

[0063] [Test Foods (Test Food and Control Food)] The test foods used were a test food containing an excipient and 250 mg of hydrolyzed globin protein per tablet (containing 0.5 w / w% of a tetrapeptide consisting of the amino acid sequence represented by SEQ ID NO: 1, manufactured by MG Pharma Co., Ltd.), and a control food containing the above excipient but without the hydrolyzed globin protein. Both the test food and the control food were yellow film-coated tablets and packaged in plain aluminum pouches so that subjects and researchers could not distinguish between the test food and the control food. The hydrolyzed globin protein was obtained by protease hydrolysis of globin protein derived from porcine red blood cells.

[0064] [Subjects] In this study, the number of subjects was designed during the planning stage. The number of subjects was determined by referring to the results of a similar study conducted prior to this study that confirmed the effect of improving liver function and preventing liver damage (Masahiro Kikuchi, Yudai Aoki, Koichi Aizawa, Hiroyuki Suganuma, Yasuhiro Nishizaki, Verification of the efficacy of a broccoli sprout extract-containing supplement on liver function in healthy adults - A multicenter randomized double-blind placebo-controlled parallel-group comparative study -, Pharmacology and Therapeutics 2018, 46(1), 81-95). Assuming an effect of 0.7 for the test food, a significance level of 0.05, and a power of 80%, it was determined that 33 subjects per group were needed, resulting in a total of 66 subjects. Assuming a dropout or discontinuation rate of approximately 20% during the study period, 82 subjects (41 in each group) were selected for this study.

[0065] Recruitment was conducted, and prospective participants visited Fukuhara Clinic under the direction of the research collaborators. The principal investigator obtained written consent from the prospective participants to participate in the study, and then conducted screening tests. Subsequently, the principal investigator reviewed the results obtained and considered the eligibility for inclusion in the study based on the following selection criteria (1) to (4) and exclusion criteria (1) to (17). As a result, it was confirmed that a sufficient number of potential subjects had been secured, and subjects to be included in the intervention (ingestion of the test food) were selected.<Selection Criteria> (1) Japanese men and women aged 20 to under 65 at the time of obtaining consent to participate in the study, (2) individuals with a blood ALT level of 20 U / L to 50 U / L at the time of screening, (3) individuals who can come to the study site on the scheduled date, (4) individuals who fully understand the purpose and content of this study and give written consent before the start of the study. <Exclusion Criteria> (1) Individuals with chronic diseases who regularly take medications; (2) Individuals with a history of serious liver disease (viral hepatitis, drug-induced liver injury, and cirrhosis), or those suspected of currently having the above liver disease; (3) Individuals with mental illness, sleep disorders, heart or kidney disease, diabetes, or other serious illnesses; (4) Individuals with a history of severe illness who are deemed unsuitable for participation in the study; (5) Individuals who may show allergic symptoms to components contained in the test food (test food and control food); (6) Shift workers or night shift workers whose work schedule extends beyond 0:00 AM; (7) Individuals with extremely irregular daily lifestyles or eating habits; (8) Individuals with a systolic blood pressure of less than 90 mmHg; (9) Individuals who, in a survey of alcohol consumption for one week starting the day after the screening test, have a habit of drinking 60g or more per day in terms of pure alcohol; (10) Individuals with a history of blood sampling Therefore, the following individuals are ineligible to participate: (11) those who have experienced malaise or deterioration in physical condition; (2) those who regularly use pharmaceuticals, quasi-drugs, herbal medicines, or health foods (including supplements) that may affect the study; (3) those who are unable to comply with requests to subjects during the study period; (4) those who are participating in another clinical trial at the time of the screening test, or who have participated in another clinical trial within two months prior to the start of the study; (5) those who have undergone apheresis donation or whole blood collection of 200 mL or more within one month prior to the screening test, or who have plans to do either during the study period; (6) those who travel or go on business trips for an average of 10 days or more per month; (7) those who wish to become pregnant during the study period, or who are pregnant (including those who may be pregnant) or breastfeeding; (8) and any other individuals deemed unsuitable by the principal investigator based on data such as health checkup results.

[0066] [Study Design] The study period was 8 weeks, and tests were conducted before the start of intake of the test food, 4 weeks after intake (after 4 weeks of intake), and 8 weeks after intake (after 8 weeks of intake).

[0067] An independent external organization, not involved in this study, acted as the allocation manager, dividing the subjects into two groups and randomly assigning them to the test food group and the control food group using random numbers. At this time, it was confirmed that there were no significant differences between the groups in the allocation factors used: age, sex, AST, ALT, and γ-GT. The allocation manager sealed the test food allocation sheet and kept it strictly stored until the end of the study, keeping the allocation confidential from all parties involved in this study, including the sponsor, principal investigator, co-investigators, consultants, and subjects, thereby ensuring the blinding of the test food.

[0068] During the study period, participants were instructed to take eight tablets of either the test food or the control food daily (2000 mg in terms of globin protein hydrolysate for the test food, or 10 mg in terms of tetrapeptide consisting of the amino acid sequence represented by SEQ ID NO: 1) with water or lukewarm water. The intake period was eight weeks, from the day before the start of intake to the day before the eight-week post-intake test. There was no specific time of intake, but it was prohibited to carry over intake to the next day if intake was not possible on the day. On the day before the start of intake and the day of the four-week post-intake test, the test food was to be taken after the test was completed.

[0069] Participants were required not to significantly change their lifestyle habits (diet, exercise, sleep, smoking, alcohol consumption, etc.) from before the study, and were prohibited from taking any medicines, quasi-drugs, herbal medicines, or health foods (including supplements) that could potentially affect the study. Blood donation was also prohibited during the study period.

[0070] [Test Items] (Serum ALT, serum AST, and serum γ-GT levels) In the screening test, pre-intake test, 4-week post-intake test, and 8-week post-intake test, serum ALT, serum AST, and serum γ-GT levels were measured as liver function markers. Serum ALT was the primary endpoint, and serum AST was the secondary endpoint.

[0071] Serum ALT, AST, and γ-GT levels were measured by blood biochemistry tests. Specifically, each was measured using the JSCC standardized method.

[0072] (Blood triglyceride levels) Blood triglyceride levels were measured during the screening test.

[0073] Blood triglyceride levels were measured by blood biochemistry tests. Specifically, they were measured using an enzymatic method.

[0074] (Daily Life Diary) From before the start of intake until the examination 8 weeks after intake, participants were asked to record daily in a daily life diary, including their wake-up and bedtime, whether or not they ate meals, the time they consumed the test food, the amount of alcohol they drank, changes in their eating and exercise habits, and the number of cigarettes they smoked.

[0075] (Physician's examination) The principal investigator conducted interviews and auscultations before the start of intake, 4 weeks after intake, and 8 weeks after intake.

[0076] [Statistical Analysis] SAS 9.4 (SAS Institute Inc.) was used for data aggregation and analysis, with a significance level of 5% (two-sided). The change in blood ALT levels at the time of screening and the time of pre-intake testing was used as a covariate, and the amount and rate of change from before the start of intake at each testing time were compared between the control food group and the test food group. Covariance analysis was performed with "time," "test food," and "time * test food" as factors, and then a t-test was performed to compare between groups.

[0077] Furthermore, to examine trends based on subject attributes, subgroup analyses were conducted for subjects with blood triglyceride levels between 120 mg / dL and 200 mg / dL in the screening test, and for subjects who consumed alcohol during the study period. Note that no correction for multiple data points was applied to the t-tests in these subgroup analyses.

[0078] [Participants in Analysis] Of the 82 subjects enrolled in this study (41 in the test food group and 41 in the control food group), one subject lost track of the study during the test food intake period, and the study was terminated at the discretion of the principal investigator. Therefore, 81 subjects completed the study. In addition, one subject was found to have not complied with the restrictions, and another subject was deemed unsuitable for analysis by the principal investigator, thus meeting the exclusion criteria. The remaining 79 subjects, excluding the one subject who lost track of the study and the two subjects who met the exclusion criteria, were included in the PPS (Per Protocol Set) analysis. One case in which abnormal changes in liver function markers were observed at the 4-week post-intake examination was excluded from the analysis at 4 weeks post-intake, as it was determined that the changes were due to the influence of medication related to COVID-19, even though the subject had no subjective symptoms.

[0079] [Results] (PPS Analysis) Table 1 shows the mean ± standard deviation of the measured blood ALT levels before ingestion, 4 weeks after ingestion, and 8 weeks after ingestion for each group included in the PPS analysis, the mean ± standard deviation of the change in blood ALT levels for each subject from before ingestion to 4 weeks after ingestion and 8 weeks after ingestion, and the mean ± standard deviation of the rate of change (%), which is the percentage of the above change relative to the blood ALT level before ingestion for each subject. The maximum blood ALT level for the PPS analysis subjects at the time of screening was 49 U / L and the minimum was 25 U / L.

[0080]

[0081] Table 2 shows the mean ± standard deviation of the measured blood AST values ​​before intake, 4 weeks after intake, and 8 weeks after intake for each group included in the PPS analysis, the mean ± standard deviation of the change in blood AST values ​​for each subject from before intake to 4 weeks after intake and 8 weeks after intake, and the mean ± standard deviation of the rate of change (%), which is the percentage of the above change relative to the blood AST value before intake for each subject. The maximum blood AST value for the PPS analysis subjects at the time of screening was 51 U / L, and the minimum value was 18 U / L.

[0082]

[0083] As shown in Table 1, the mean percentage change in blood ALT levels in the test food group was negative and smaller than that of the control food group at both 4 weeks and 8 weeks after intake. Therefore, it was shown that intake of the tetrapeptide consisting of the amino acid sequence represented by Sequence ID No. 1 can improve blood ALT levels in healthy individuals.

[0084] Furthermore, as shown in Table 2, the mean percentage change in blood AST levels in the test food group was negative and smaller than that of the control food group at both 4 weeks and 8 weeks after intake. Therefore, it was shown that intake of the tetrapeptide consisting of the amino acid sequence represented by Sequence ID No. 1 can improve blood AST levels in healthy individuals.

[0085] (Subgroup analysis 1: Subjects with blood triglyceride levels of 120 mg / dL or higher and less than 200 mg / dL) Table 3 shows the mean ± standard deviation of the measured blood ALT and AST values ​​before ingestion, 4 weeks after ingestion, and 8 weeks after ingestion, the mean ± standard deviation of the change in blood ALT and AST values ​​for each subject from before ingestion to 4 weeks after ingestion and 8 weeks after ingestion, and the mean ± standard deviation of the rate of change (%), which is the percentage change for each subject relative to the blood ALT and AST values ​​before ingestion, for each subgroup of subjects in the PPS analysis whose blood triglyceride levels in the above screening test were 120 mg / dL or higher and less than 200 mg / dL.

[0086]

[0087] As shown in Table 3, at both 4 weeks and 8 weeks after ingestion, the mean percentage changes in blood ALT and blood AST levels in the test food group were negative and smaller than those in the control food group. Furthermore, at 8 weeks after ingestion, the percentage changes in blood ALT and blood AST levels in the test food group were significantly smaller than those in the control food group. Also, for the test food group with blood triglyceride levels between 120 mg / dL and 200 mg / dL, the mean percentage changes in blood ALT levels after 4 weeks, and the mean percentage changes in blood ALT and blood AST levels after 8 weeks, were both smaller than those of PPS. Therefore, it was shown that when individuals with blood triglyceride levels between 120 mg / dL and 200 mg / dL ingested the tetrapeptide consisting of the amino acid sequence represented by Sequence ID No. 1, their blood ALT and blood AST levels improved more effectively, and this trend was particularly pronounced when the tetrapeptide was ingested for 8 weeks or more.

[0088] (Subgroup Analysis 2: Subjects who consumed alcohol during the intake period) Table 4 shows the mean ± standard deviation of the measured blood ALT and blood AST values ​​before intake, 4 weeks after intake, and 8 weeks after intake, the mean ± standard deviation of the change in blood ALT and blood AST values ​​for each subject from before intake to 4 weeks after intake and 8 weeks after intake, and the mean ± standard deviation of the percentage change (%) of the above change relative to the blood ALT and blood AST values ​​for each subject, for each subgroup of the PPS analysis subjects who consumed alcohol at least once during the intake period. Note that the amount of pure alcohol consumed during the intake period (8 weeks) in the test food group that consumed alcohol was between 0.4 g / day and 42.2 g / day.

[0089]

[0090] As shown in Table 4, at both 4 weeks and 8 weeks after ingestion, the mean percentage changes in blood ALT and AST levels in the test food group were negative and smaller than those in the control food group. Furthermore, at 8 weeks after ingestion, the percentage changes in blood ALT and AST levels in the test food group were significantly smaller than those in the control food group. Additionally, the mean percentage changes in blood ALT and AST levels at 4 weeks and 8 weeks after ingestion for test food groups that consumed alcohol at least once during the ingestion period were both smaller than those in the PPS group. Moreover, as mentioned above, participants were instructed not to significantly change their lifestyle, such as drinking alcohol, during the ingestion period; therefore, subjects who consumed alcohol at least once during the ingestion period can be considered to have a drinking habit. Therefore, it was shown that when individuals with a drinking habit ingested the tetrapeptide consisting of the amino acid sequence represented by Sequence ID No. 1, their blood ALT and AST levels improved more effectively, and this trend was particularly pronounced when the tetrapeptide was ingested for eight weeks or more.

Claims

1. A composition for use in healthy humans for improving, maintaining, and / or preventing deterioration of liver function, comprising a tetrapeptide consisting of the amino acid sequence represented by Sequence ID No.

1.

2. A composition for healthy human use that contains a tetrapeptide consisting of the amino acid sequence represented by Sequence ID No. 1, for improving, maintaining, and / or preventing deterioration of blood alanine aminotransferase (ALT) levels or blood aspartate aminotransferase (AST) levels.

3. The composition according to claim 1 or 2, wherein the composition contains a globin protein hydrolysate, and at least a portion of the tetrapeptide is derived from the globin protein hydrolysate.

4. The composition according to claim 1 or 2, wherein the person is a person whose blood triglyceride level is 120 mg / dL or more and less than 200 mg / dL.

5. The composition according to claim 1 or 2, wherein the person is a person who drinks alcohol.

6. The composition according to claim 1 or 2, wherein the person is a person whose blood ALT level is 20 U / L or more and 50 U / L or less.

7. The composition according to claim 1 or 2, which is used to be ingested or administered in an amount of 5 mg or more and 15 mg or less per day in terms of tetrapeptide.

8. The composition according to claim 7, which is used to be ingested or administered for four weeks or more.

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