JP7134322B2 - Composition for suppressing AGEs formation - Google Patents

Description

TECHNICAL FIELD The present invention relates to an AGE formation inhibitory composition, and more particularly to an AGE formation inhibitor, especially an AGE formation inhibitor for external use.

AGEs (abbreviation for Advanced Glycation End Products) is a general term for products produced by the glycation reaction of proteins, and is called advanced glycation products or advanced glycation products. It has been reported that AGEs accumulate in the body and cause or exacerbate various symptoms related to age-related chronic diseases such as diabetes, arteriosclerosis, and neurological diseases. ing.

FIG. 1 is a cross-sectional view schematically showing the hair growth mechanism. Dermal papilla cells at the root of hair deliver nutrients and hair growth factors to hair matrix cells to grow hair. Hair matrix cells are cells that surround dermal papilla cells and actively undergo cell division to form hair. Therefore, hair growth is maintained by firmly receiving hair growth factors in hair matrix cells. Conversely, if the cooperative relationship between the dermal papilla cells and the hair matrix cells does not go well, the hair becomes thin.

Our recent research (Non-Patent Document 1) has revealed that when AGEs accumulate in dermal papilla cells, inflammatory substances that upset the cooperative relationship between the two cells are released from dermal papilla cells. As a result, cell division of hair matrix cells is disturbed and hair growth is suppressed.

Eur J Dermatol. 2015 Jul-Aug;25(4):359-61 Matsui T, Yamagishi S, et al. Immunol Lett. 2015 Oct;167(2):141-6

In such cases, the hair becomes finer and thinner overall, so AGEs seem to be related to the diffuse thinning of hair on the top of the head, which is more common in women. In order for AGEs to act on dermal papilla cells, AGEs must get stuck in the keyholes that exist in the cells. The holes in which AGEs get stuck are called AGE receptors, and are called RAGE in English. In other words, AGEs were involved not only in wrinkles and sagging, but also in thinning hair.

The present invention has been made to solve the above problems, and an object of the present invention is to provide a composition for suppressing the formation of AGEs which is excellent in safety and usefulness.

The present invention provides a composition for suppressing AGE formation containing celery extract and rutin as active ingredients. A composition containing celery extract and rutin may be hereinafter referred to as "CR2".

In one form, the celery extract is an aqueous extract of celery.

In one embodiment, the ratio of the weight of celery to the volume of water is 1/5 to 1/0.2.

In one aspect, the celery comprises celery stalks.

The present invention also provides an AGE formation inhibitor containing any of the above compositions for inhibiting AGE formation.

The present invention also provides the above-mentioned AGE formation inhibitor, which is an external preparation.

The present invention also provides the above AGE formation inhibitor, which is a shampoo, rinse or hair restorer.

INDUSTRIAL APPLICABILITY According to the present invention, a composition for suppressing AGE formation that is excellent in safety and usefulness is provided.

FIG. 4 is a cross-sectional view schematically showing the hair growth mechanism. 1 is a graph showing how the amount of AGEs produced from albumin and glyceraldehyde changes in the presence of celery extract, rutin, or both. 1 is a graph showing how albumin-induced oxidative stress levels in human endothelial cells change in the presence of celery extract. Fig. 3 is a graph showing how the RAGE gene expression level by albumin in human endothelial cells changes in the presence of celery extract.

<Celery extract>
"Celery", whose scientific name is "Apium graveolens var. dulce", is a plant of the Umbelliferae family. Celery is classified as a light-colored vegetable, and is roughly divided into four varieties: intermediate, green, yellow, and native oriental varieties. In the present invention, celery stems, leaves, or stems and leaves are used. Celery extract refers to a liquid component obtained by removing water-insoluble components from celery stems or leaves.

A celery extract can be produced by preparing celery as a raw material, pulverizing, squeezing, filtering, solid-liquid separation, and, if necessary, purification. A solvent may be added to the crushed celery during juicing. As a result, the liquid component can be efficiently extracted from the celery.

As a solvent for extracting liquid components from celery, water, 1,3-butylene glycol, or other organic solvents can be used (including water containing organic solvents). The organic solvent typically includes methanol and ethanol, but widely includes known organic solvents such as acetone, diethyl ether, ethyl acetate, and hexane. When water is used as the extraction solvent, a phosphate buffer is preferably used from the viewpoint of stabilizing the pH of the resulting celery extract.

For example, a celery extract can be obtained by adding a predetermined amount of water to a predetermined amount of celery, pulverizing the celery with a mixer, filtering and centrifuging, and then collecting the clear supernatant. The amounts of celery and water used are, for example, such that the ratio of the weight of ground celery to the volume of water is 1/5 to 1/0.2. By setting the amounts of celery and water used within this range, the stability of the obtained celery extract is improved. The ratio of the weight of ground celery to the volume of water is preferably 1/2 to 1/0.5, more preferably 1/1.5 to 1/0.7.

As a source of celery extract, concentrated celery juice or reduced celery juice may be used, thereby eliminating the steps of preparing, crushing and squeezing the celery. You may use the celery extract marketed as a quasi-drug.

The amount of celery extract to be used is 0.2 to 5.0% by weight based on the weight of the composition for suppressing AGEs formation. By setting the amount of celery extract to be used within the above range, the effect of suppressing the formation of AGEs is improved. The amount of celery extract used is preferably 0.4 to 3.0% by weight, more preferably 0.7 to 2.0% by weight, based on the volume of the AGEs formation-suppressing composition.

<Rutin>
Rutin is one of the tasteless and odorless flavonoids widely contained in higher plants, vegetables and fruits. It has attracted attention as a natural extract with many functions including antioxidant action, and is a substance with a proven track record in applications as pharmaceuticals and raw materials for pharmaceuticals.

Rutin can be produced, for example, by extracting it from a rutin-rich plant such as buckwheat sprouts. For rutin, for example, commercially available products can be used. Rutin may be rutin that has been modified to have increased water solubility. A specific example of such water-soluble rutin is α-glucosylated rutin.

The amount of rutin used is 20 to 500 μM (micromoles/liter) based on the volume of the composition for suppressing AGE formation. By setting the amount of rutin used within the above range, the effect of suppressing AGE formation is improved. The amount of rutin used is preferably 50-250 μM, more preferably 80-150 μM, based on the volume of the composition for suppressing AGE formation.

<Composition for suppressing AGE formation>
INDUSTRIAL APPLICABILITY The composition for suppressing AGE formation of the present invention exhibits an excellent AGE formation suppressing effect as an AGE formation inhibitor. The composition for suppressing AGE formation of the present invention, for example, not only suppresses the formation of AGEs in the scalp, but also supports hair growth by suppressing endothelial cell damage and inflammatory reaction caused by AGEs already present in the scalp and dermal papilla. . The composition for suppressing formation of AGEs of the present invention is extremely useful when it is used as an external preparation by being contained in soaps, cosmetics, etc., especially when it is contained in shampoos, rinses, hair restorers, etc. and used as an external preparation. high.

<Regarding the action of CR2 (celery extract + rutin)>
Using celery extract and rutin, two ingredients that are quasi-drugs and are recognized as medicinal soaps, shampoos and conditioners, hair removers, hair growth agents, other medicated cosmetics, armpit odor inhibitors, contraindications, and bath agents. , AGEs formation inhibitory effect, oxidative stress production by AGEs and RAGE gene expression inhibitory effect were investigated. First, 50 ml of phosphate buffer was added to 50 g of fresh celery, pulverized with a mixer, filtered with a case, and centrifuged. Rutin was purchased from Wako Pure Chemical with a purity of 90%.

1) Inhibition of AGE formation 25 mg/ml bovine serum albumin and 2 mM glyceraldehyde (reducing sugar) in 0.2 M phosphate buffer (pH 7.4) solution, final concentration 1% celery extract, final concentration 100 μM rutin present. , incubated at 37°C for 1 day in the absence. After that, the sample was dialyzed against a phosphate buffer, and the amount of albumin converted to AGEs was measured by an ELISA method using a monoclonal antibody against AGEs as previously reported (Reference: Non-Patent Document 2). The 50% inhibitory concentration of albumin converted to AGEs in the standard curve was defined as 1U.

Results: When AGEs derived from glyceraldehyde were specifically measured by ELISA, administration of rutin and celery extract alone tended to suppress the formation of AGEs, but the effect was statistically significant. was not significant for However, it was found that the formation of AGEs was significantly suppressed in the presence of a final concentration of 1% celery extract and a final concentration of 100 μM rutin (Fig. 2, N = 5 for each group, results are mean ± standard deviation. Statistics are analyzed by Student's t test.*, p<0.05).

2) Effect on oxidative stress production in endothelial cells and expression induction of RAGE gene, which is an AGE receptor After treatment with control albumin (hereinafter referred to as albumin), oxidative stress production was analyzed 30 minutes later, and RAGE gene expression was analyzed 4 hours later by the fluorescence reagent method and RT-PCR method, respectively.

Results: It was clarified that 1% celery extract suppresses the production of oxidative stress by AGEs and the induction of RAGE gene expression in endothelial cells (Figures 3 and 4; Expression: N = 5 in each group, results are mean ± standard deviation Statistics were analyzed by Student's t test *, p<0.05, **, p<0.01, albumin + 2 mM glyceraldehyde (1 day)) .

<Discussion>
In this study, it was clarified for the first time in the world that the combination of two quasi-drugs, celery extract and rutin (CR2), significantly suppressed the formation of AGEs. Furthermore, celery extract was shown to reduce reactivity to AGEs in endothelial cells through antioxidant and RAGE expression suppression effects. From the above, CR2 not only suppresses the formation of AGEs in the scalp, but also suppresses endothelial cell damage and inflammatory reactions caused by AGEs already present in the scalp and dermal papilla, thereby supporting hair growth. . Considering the adverse effects of AGEs-RAGE on beauty, such as wrinkles, spots, and thinning hair, CR2 is expected to be a combination of ingredients that are extremely useful as shampoos, conditioners, and hair restorers.

Claims (12)

A composition for inhibiting AGE formation containing celery extract and rutin as active ingredients,
The celery extract comprises a celery stem, leaf, or stem and leaf extract,
The amount of rutin used is 80 to 150 μM based on the volume of the composition for suppressing AGEs formation ,
A composition for suppressing the formation of AGEs by suppressing the reaction between protein and sugar (rutin excludes rutin contained in celery extract). 2. The composition for inhibiting AGEs formation according to claim 1, wherein the celery extract is an extract of celery with water, 1,3-butylene glycol, or another organic solvent (including water containing an organic solvent). 3. The composition for inhibiting AGEs formation according to claim 1, wherein the celery extract is an extract of celery with water or 1,3-butylene glycol (including water containing 1,3-butylene glycol). The composition for suppressing AGEs formation according to any one of claims 1 to 3, wherein the ratio of the weight of celery to the volume of extracted water is 1/5 to 1/0.2. The composition for suppressing AGEs formation according to any one of claims 1 to 4, wherein the celery contains a celery stem. The composition for suppressing AGEs formation according to any one of claims 1 to 5, wherein the amount of the celery extract used is 0.2 to 5.0% by weight based on the weight of the composition for suppressing AGEs formation. . The composition for suppressing AGEs formation according to any one of claims 1 to 6, wherein the amount of rutin used is 100 µM based on the volume of the composition for suppressing AGEs formation. An AGEs formation inhibitor comprising the composition for inhibiting AGEs formation according to any one of claims 1 to 7 . 9. The AGE formation inhibitor according to claim 8 , which is an external preparation. 9. The AGE formation inhibitor according to claim 8 , which is a shampoo, rinse or hair restorer. providing a celery extract;
providing rutin and combining the celery extract and rutin;
A method for producing a composition for suppressing AGEs formation, comprising
The celery extract comprises a celery stem, leaf, or stem and leaf extract,
The amount of rutin used is 80 to 150 μM based on the volume of the composition for suppressing AGEs formation ,
A method for producing a composition for suppressing the formation of AGEs by suppressing the reaction between protein and sugar (rutin excludes rutin contained in celery extract) . 12. The method for producing a composition for suppressing AGEs formation according to claim 11, wherein the amount of rutin used is 100 [mu]M based on the volume of the composition for suppressing AGEs formation.

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