Composition for promoting expression of glutamate cysteine ligase catalytic subunit (GCLC) gene

Abstract

To provide a composition for promoting the expression of a glutamate cysteine ligase catalytic subunit (GCLC) gene.SOLUTION: A composition for promoting the expression of a glutamate cysteine ligase catalytic subunit (GCLC) gene contains one or more components selected from the group consisting of jujube, ginger, and platycodon root.SELECTED DRAWING: None

Description

[Technical field]

[0001] The present invention relates to a composition for promoting expression of the glutamate cysteine ​​ligase catalytic subunit (GCLC) gene. [Background technology]

[0002] Glutathione is an antioxidant that is widely distributed in the body. It is a tripeptide consisting of cysteine, glutamic acid, and glycine. As the main reducing agent in the body, it plays an important role in various redox metabolic processes, such as cell defense, xenobiotic metabolism, and repair processes.

[0003] Glutathione exists in two forms: reduced glutathione (GSH) and oxidized glutathione (GSSG), in which the side chain thiol groups of cysteine ​​are bonded together via disulfide bonds. In the body, more than 98% exists as reduced glutathione due to the action of glutathione reductase, and therefore glutathione acts to reduce free radicals and peroxides in the body and protects cells. In addition, because the side chain thiol group of glutathione is nucleophilic, it also attacks and detoxifies toxic compounds.

[0004] Normally, when cysteine, an amino acid that constitutes proteins, exists alone in the body, it reacts with itself to become cystine, but this reaction generates radicals, which keeps the intracellular cysteine ​​concentration relatively low. On the other hand, glutathione is broken down into each of the constituent amino acids by γ-glutamyltranspeptidase and dipeptidase, and is thought to function as a cysteine ​​source in the body.

[0005] Therefore, it is believed that when glutathione in the body is reduced and normal cell function cannot be maintained, various disorders are caused.In fact, it has been reported that the decrease in intracellular concentration of glutathione causes cell damage, inflammation, blackening, age spots, freckles, acute or chronic alcoholic liver disease, liver disease, chronic renal failure, lung disease caused by smoking, idiopathic pulmonary fibrosis, cataract, ischemic heart disease, Parkinson's disease, Alzheimer's disease, gastric ulcer, adult respiratory syndrome, immunodeficiency, bone marrow hypoplasia, acquired immune deficiency syndrome, latent viral infection, and aging phenomenon and carcinogenesis associated with physiological aging.

[0006] In order to prevent or improve such disorders, research has been conducted on the use of glutathione in the fields of medicine and beauty, and various useful effects of glutathione, such as skin-whitening effects and liver function improvement effects, have been reported. On the other hand, there are problems with using glutathione itself as a drug, cosmetic, or quasi-drug, such as the Pharmaceutical and Medical Device Act and the fact that glutathione itself is relatively chemically unstable. Therefore, various safe and continuously ingestible ingredients that can promote the production of glutathione in the body have been studied.

[0007] For example, a composition for enhancing glutathione containing a plant such as meadowsweet (Patent Document 1), a glutathione production promoter containing luteolin or its glycoside as an active ingredient (Patent Document 2), etc. have been reported.

[0008] Thus, various drugs have been found to promote glutathione production, but their effects are not always sufficient, and there is still a demand for drugs that promote glutathione production with better effects. [Prior art documents] [Patent documents]

[0009] [Patent Document 1] JP 2002-275079 A [Patent Document 2] JP 2022-187468 A Summary of the Invention

[0010] The present invention provides a composition for promoting expression of the glutamate cysteine ​​ligase catalytic subunit (GCLC) gene.

[0011] The present inventors have found that one or more components selected from the group consisting of jujube, ginger, and Chinese bellflower are useful for promoting the expression of glutamate cysteine ​​ligase catalytic subunit (GCLC) gene. The present invention is based on this finding.

[0012] According to the present invention, the following inventions are provided. (1) A composition for promoting expression of a glutamate cysteine ​​ligase catalytic subunit (GCLC) gene, comprising one or more components selected from the group consisting of jujube, ginger, and bellflower. (2) A composition for promoting the activity of glutathione synthetase, comprising one or more components selected from the group consisting of jujube, ginger, and bellflower. (3) A composition for promoting glutathione production comprising one or more components selected from the group consisting of jujube, ginger, and bellflower. (4) The composition according to any one of (1) to (3) for improving skin transparency. (5) A composition according to any one of (1) to (4) for promoting the production of pheomelanin. (6) The composition according to any one of (1) to (5), which is a food composition. (7) Use of one or more components selected from the group consisting of jujube, ginger, and bellflower for the manufacture of a drug intended to promote expression of a glutamate cysteine ​​ligase catalytic subunit (GCLC) gene. (8) Use of one or more components selected from the group consisting of jujube, ginger, and bellflower for the manufacture of a medicament for promoting the activity of glutathione synthetase. (9) Use of one or more components selected from the group consisting of jujube, ginger, and bellflower for the manufacture of a medicament for promoting glutathione production. (10) The use according to any one of (7) to (9) for the manufacture of a drug for improving skin transparency. (11) The use according to any one of (7) to (10) for the manufacture of a drug intended to promote the production of pheomelanin. (12) Use of a composition comprising one or more components selected from the group consisting of jujube, ginger, and bellflower, for promoting expression of the glutamate cysteine ​​ligase catalytic subunit (GCLC) gene. (13) Use of a composition comprising one or more components selected from the group consisting of jujube, ginger, and bellflower for promoting the activity of glutathione synthetase. (14) Use of a composition comprising one or more ingredients selected from the group consisting of jujube, ginger, and bellflower for promoting glutathione production. (15) The use according to any one of (12) to (14) for improving skin transparency. (16) The use according to any one of (12) to (15) for promoting the production of pheomelanin. (17) Jujube, ginger, or Chinese bellflower to promote expression of the glutamate cysteine ​​ligase catalytic subunit (GCLC) gene. (18) Jujube, ginger, or bellflower to promote glutathione synthetase activity. (19) Jujube, ginger, or bellflower to promote glutathione production. (20) The jujube, ginger, or bellflower according to any one of (17) to (19) for improving skin transparency. (21) The jujube, ginger, or bellflower according to any one of (17) to (20) for promoting the production of pheomelanin. (22) A method for promoting expression of the glutamate cysteine ​​ligase catalytic subunit (GCLC) gene in a subject, the method comprising administering an effective amount of one or more components selected from the group consisting of jujube, ginger, and platycodon. (23) A method for promoting glutathione synthetase activity in a subject, comprising administering an effective amount of one or more ingredients selected from the group consisting of jujube, ginger, and platycodon. (24) A method for promoting glutathione production in a subject, comprising administering an effective amount of one or more ingredients selected from the group consisting of jujube, ginger, and platycodon. (25) A method for improving the transparency of the skin of a subject, comprising administering an effective amount of one or more ingredients selected from the group consisting of jujube, ginger, and bellflower. (26) A method for promoting pheomelanin production in a subject, comprising administering an effective amount of one or more components selected from the group consisting of jujube, ginger, and bellflower.

[0013] According to the present invention, a composition for promoting the expression of glutamate cysteine ​​ligase catalytic subunit (GCLC) gene is provided. According to the present invention, a composition for promoting the activity of glutathione synthetase can be provided. According to the present invention, a composition for promoting the production of glutathione can be provided. [Brief description of the drawings]

[0014] [Figure 1] FIG. 1 shows the results of evaluating the effect of Zizyphus jujube on the expression of the glutamate cysteine ​​ligase catalytic subunit (GCLC) gene in epidermal keratinocytes. [Diagram 2] FIG. 2 shows the results of evaluating the effect of ginger on the expression of the glutamate cysteine ​​ligase catalytic subunit (GCLC) gene in epidermal keratinocytes. [Diagram 3] FIG. 3 shows the results of evaluating the effect of Platycodon grandiflorum on the expression of glutamate cysteine ​​ligase catalytic subunit (GCLC) gene in epidermal keratinocytes. Description of the Invention

[0015] According to one embodiment of the present invention, a composition for promoting the expression of glutamate cysteine ​​ligase catalytic subunit (GCLC) gene is provided, the composition comprising one or more components selected from the group consisting of jujube, ginger, and Chinese bellflower. Such a composition may comprise a GCLC gene expression promoter, and the GCLC gene expression promoter may comprise or consist of one or more components selected from the group consisting of jujube, ginger, and Chinese bellflower.

[0016] γ-Glutamylcysteine ​​is a precursor of glutathione, which is a peptide bond between glutamic acid and cysteine, and is biosynthesized from glutamic acid and cysteine ​​by γ-glutamylcysteine ​​synthetase (γGCS). γGCS is the rate-limiting enzyme in glutathione biosynthesis and is known to consist of two subunits, glutamic acid cysteine ​​ligase modifying subunit (GCLM) and glutamic acid cysteine ​​ligase catalytic subunit (GCLC).

[0017] Therefore, according to another aspect of the present invention, there is provided a composition for promoting the activity of glutathione synthetase, the composition comprising one or more components selected from the group consisting of jujube, ginger, and Chinese bellflower. Such a composition may comprise a glutathione synthetase activity promoter, and the glutathione synthetase activity promoter may comprise or consist of one or more components selected from the group consisting of jujube, ginger, and Chinese bellflower.

[0018] The glutathione synthetase in the present invention may be any enzyme that is directly or indirectly involved in the biosynthesis of glutathione, and is preferably γ-glutamylcysteine ​​synthetase (γGCS).

[0019] According to yet another embodiment of the present invention, there is provided a composition for promoting glutathione production, the composition comprising one or more components selected from the group consisting of jujube, ginger, and Chinese bellflower. Such a composition may comprise a glutathione production promoter, and the glutathione production promoter may consist of or comprise one or more components selected from the group consisting of jujube, ginger, and Chinese bellflower.

[0020] In general, melanin is produced by the oxidation of tyrosine by copper ion-dependent tyrosinase in the melanosomes of melanocytes to produce dihydroxyphenylalanine (DOPA), which is then converted to dopaquinone by DOPA-oxidase, and then polymerized nonenzymatically in melanocytes to dopachrome and indolequinone. The melanin produced in this way is called eumelanin (true melanin) and is black-brown in color. In addition to true melanin, orange-red pheomelanin (submelanin), which is synthesized from dopaquinone and cysteine ​​via dihydrobenzothiazine, is also known as melanin (see Pigment Cell Melanoma Res, 28(5), 520-44(2015), etc.).

[0021] It has been reported that glutathione not only inhibits the production of eumelanin by binding to tyrosinase and inhibiting its activity, but also has the specificity of binding to dopaquinone to produce pheomelanin (see, for example, Yakugaku Zasshi, 128(8), 1203-7(2008); Postepy Hig Med Dosw, 70(0), 695-708(2016)).

[0022] Therefore, according to a preferred embodiment of the present invention, the composition of the present invention is a composition for promoting the production of pheomelanin.

[0023] The skin color of human skin is composed of a balance of eumelanin (blackish brown) and pheomelanin (orange-red), and this balance also has a great influence on the brightness of the skin. For example, it has been reported that, when comparing melanocytes derived from a human with dark skin with melanocytes derived from a human with light skin, more pheomelanin is produced in the melanocytes derived from the human with light skin (see, for example, The Journal of investigative dermatology, 122, 1251-1255 (2004)). It is believed that promoting the production of pheomelanin, etc., can lead to a constitution in which pheomelanin is dominant over eumelanin, thereby contributing to the maintenance and improvement of skin brightness, and glutathione has a great influence on the promotion of this pheomelanin production. Therefore, according to another preferred embodiment of the present invention, the composition of the present invention is a composition for maintaining and / or improving skin brightness.

[0024] In addition, skin brightness can affect skin transparency, just like other factors such as melanin amount, complexion, hemoglobin amount, hemoglobin oxygen saturation, skin texture, etc. Therefore, according to another preferred embodiment of the present invention, the composition of the present invention is a composition for maintaining and / or improving skin transparency.

[0025] In the present invention, "jujube" refers to the dried fruit of jujube (Zizyphus jujuba Miller var. inermis Rehder), a deciduous tree of the Rhamnaceae family, which is also used as a herbal medicine in China and other places.

[0026] The form of the "jujube" in the present invention is not limited as long as it has the desired effect of the present invention. For example, powdered form, extract, extract powder, etc. can be used.

[0027] In the present invention, "ginger" refers to the rhizome of ginger (Zingiber officinale), a perennial plant of the Zingiberaceae family, and may be dried. In particular, ginger rhizomes that have been parboiled or steamed and dried are called "dried ginger." Both "ginger" and "dried ginger" are used as herbal medicines in China and other countries.

[0028] The form of "ginger" in the present invention is not limited as long as it has the intended effect of the present invention. For example, powdered ginger, extract, extract powder, etc. can be used.

[0029] In the present invention, "bellflower" refers to Platycodon grandiflorum A. De Candolle (sometimes called Campanulaceae) of the Campanulaceae family. The part to be used is not limited, but the root is preferred. It is also used as a herbal medicine in China and other places.

[0030] The form of the "Platycodon grandiflorum" in the present invention is not limited as long as it has the desired effect of the present invention. For example, powdered form, extract, extract powder, etc. can be used.

[0031] The method for producing the extract of jujube, ginger, or bellflower in the present invention is not particularly limited, and can be produced according to a method commonly used in the art, for example, by extracting the raw material of the extract. The extraction method is not limited, but includes, for example, hot water extraction, ultrasonic extraction, filtration, reflux extraction, solvent (e.g., ethanol) extraction, etc. These may be carried out alone, or two or more methods may be used in combination. In addition, the extract may be further extracted one or more times in the same manner to obtain a high-purity extract. In addition, such an extract may be made into extract powder by carrying out an appropriate process such as powderization by spray drying.

[0032] The type of solvent used for producing the extract of jujube, ginger, or bellflower in the present invention is not particularly limited, and any solvent known in the art may be used as long as it can produce an extract having the desired effect of the present invention. Examples of such solvents include, but are not limited to, water, alcohols having 1 to 4 carbon atoms, ethyl acetate, acetone, chloroform, etc., and two or more of these may be used in combination.

[0033] If jujube, ginger, or platycodon (including powdered forms, extracts, extract powders, etc.) are commercially available, they may be used as long as they have the desired effect of the present invention.

[0034] The composition of the present invention may be applied either in vivo or in vitro.

[0035] When the composition of the present invention is applied to a subject, examples of the subject to which it can be applied include, but are not limited to, humans, primates including chimpanzees, pet animals such as dogs and cats, livestock animals such as cows, horses, sheep and goats, and mammals such as rodents such as mice and rats.

[0036] When the composition of the present invention is applied to a biological sample, the biological sample to be applied includes, but is not limited to, cells obtained from a living body, their passaged cells, or established cell lines, and these cells may be differentiated and used as necessary. Such cells may be cells derived from any animal or plant, and are, but are not limited to, preferably animal cells, and preferably human cells.

[0037] The amount of jujube used in the composition of the present invention can be appropriately determined depending on the application, but preferably, the dry weight of jujube is 1 mg to 15 g, more preferably 3 mg to 5 g, and even more preferably 10 mg to 1.5 g, relative to the total amount of the composition.

[0038] The amount of ginger used in the composition of the present invention can be appropriately determined depending on the application, but preferably, the dry weight of ginger is 1 mg to 10 g, more preferably 3 mg to 0.33 g, and more preferably 10 mg to 1 g, relative to the total amount of the composition.

[0039] The amount of Platycodon grandiflorum used in the composition of the present invention can be appropriately determined depending on the application, but preferably, the dry weight of Platycodon grandiflorum is 1 mg to 10 g, more preferably 3 mg to 0.33 g, and more preferably 10 mg to 1 g, relative to the total amount of the composition.

[0040] The composition of the present invention may be an oral composition or a topical composition.The jujube, ginger and bellflower of the present invention are preferably absorbed by the digestive tract when taken orally, and their active ingredients are believed to act in the body.Therefore, according to one preferred embodiment of the present invention, the composition of the present invention is an oral composition.

[0041] When the composition of the present invention is used as an oral composition, it can further contain known components (additives) that are used in oral compositions.Such components include, but are not limited to, emulsifiers, hydrating agents, solvents, emollients, stabilizers, thickeners, preservatives, lubricants, chelating agents, fillers, excipients, powders, fragrances, flavors, absorbents, dyes, opacifiers, antioxidants, preservatives, vitamins, amino acids, nutrients, minerals (electrolytes), flavors such as synthetic flavors and natural flavors, pectinic acid and its salts, alginic acid and its salts, organic acids, pH regulators, glycerin, alcohol, carbonation agents used in carbonated drinks, etc.

[0042] The oral composition of the present invention may be any of liquid, semi-solid, and solid.

[0043] When the composition of the present invention is used as a topical composition, it can further contain known components (additives) that are used in topical compositions.Such components include, but are not limited to, emulsifiers, hydrating agents, solvents, emollients, stabilizers, thickeners, preservatives, lubricants, chelating agents, fillers, excipients, powders, fragrances, flavorings, absorbents, dyes, opacifiers, antioxidants, vitamins, amino acids, etc.

[0044] The topical compositions of the present invention may be liquid, semi-solid, or solid.

[0045] The composition of the present invention may be any of a pharmaceutical composition, a cosmetic composition, and a food composition, and is preferably a food composition. EXAMPLES

[0046] The present invention will be described in detail with reference to the following examples, but the present invention is not limited to these examples. Contents are expressed in mass % unless otherwise specified.

[0047] Extraction of Jujube A 10-fold amount of purified water was added to the cut-up herbal medicine jujube, and the mixture was heated to 95-100°C while stirring, and then extracted for about 60 minutes. After the extraction was completed, the extract was separated into solid and liquid, and the separated liquid was concentrated under reduced pressure and then spray-dried to obtain a dried jujube extract.

[0048] Ginger Extract The cut raw ginger was mixed with 10 times the amount of purified water, heated to 95-100°C while stirring, and extracted for about 60 minutes. After the extraction, the extract was separated into solid and liquid, concentrated under reduced pressure, and then spray-dried to obtain a dried ginger extract.

[0049] Platycodon Extract The cut herbal medicine Platycodon grandiflorum was mixed with 10 times the amount of purified water, heated to 95-100°C while stirring, and extracted for about 60 minutes after the temperature was raised. After the extraction was completed, the extract was separated into solid and liquid, and the separated liquid was concentrated under reduced pressure and then spray-dried to obtain a dried Platycodon grandiflorum extract.

[0050] Evaluation of the effect of drugs (each extract) on GCLC gene expression in normal human epidermal keratinocytes Normal human epidermal keratinocytes (passage number: 4, Kuraray Industries, Inc.) were cultured in a 96-well plate at 1.5 × 10 4 The cells were seeded at a cell density of 10 ... Real-time PCR was performed using the Step OnePlus™ Real-Time PCR System. Analysis was performed using the delta-delta CT method, and the gene expression levels of each group were expressed as relative values, with the correction value of the test drug untreated group (control) set at 1. After calculating the values, unpaired Student's t-test was performed as a statistical treatment.

[0051] The results of evaluating the effect of jujube on GCLC gene expression are shown in Figure 1, the effect of ginger on GCLC gene expression in Figure 2, and the effect of bellflower on GCLC gene expression in Figure 3. In Figure 1, a significant increase in GCLC gene expression was observed in the group to which 20 μg / mL of jujube was added compared to the control group. In addition, in Figure 2, a significant increase in GCLC gene expression was observed in the group to which 0.2 μg / mL of ginger was added compared to the control group. In addition, in Figure 3, a significant increase in GCLC gene expression was observed in the group to which 2 μg / mL of bellflower was added compared to the control group. These results suggest that the three components of jujube, ginger, and bellflower promote the expression of GCLC gene.

Claims

1. A composition for promoting expression of a glutamate cysteine ​​ligase catalytic subunit (GCLC) gene, comprising one or more components selected from the group consisting of jujube, ginger, and bellflower.

2. A composition for promoting the activity of glutathione synthetase, comprising one or more components selected from the group consisting of jujube, ginger, and bellflower.

3. A composition for promoting glutathione production comprising one or more components selected from the group consisting of jujube, ginger, and bellflower.

4. The composition according to any one of claims 1 to 3 for improving skin clarity.

5. The composition according to any one of claims 1 to 3 for promoting the production of pheomelanin.

6. The composition according to any one of claims 1 to 3, which is a food composition.

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