JP2021035923A - Saccharification reaction inhibitory composition - Google Patents

Abstract

To provide an in vivo protein saccharification reaction inhibitory composition.SOLUTION: An in vivo protein saccharification reaction inhibitory composition contains extract from Salicornia europaea as an active ingredient. The protein is a skin horny layer protein. The extract from Salicornia europaea is extract obtained from a whole plant of Salicornia europaea by a supercritical method.SELECTED DRAWING: Figure 2

Description

The present invention relates to a composition that suppresses a protein glycation reaction in a living body.

Glycation is a phenomenon in which reducing sugars and proteins are bound by a non-enzymatic irreversible reaction (Maillard reaction) to form advanced glycation end products (AGEs) via various intermediates.
With the aging of the living body, this Maillard reaction occurs in each tissue and site in the body, and AGEs accumulate. As the glycation reaction progresses in the skin, AGEs accumulate in the skin tissue. Accumulation of AGEs on the skin causes yellowing of the skin and a decrease in transparency, which is not preferable from the viewpoint of cosmetics.

In order to suppress this saccharification reaction, a method of suppressing protein saccharification by applying a substance that suppresses saccharification from the skin surface is known. Patent Document 1 describes that, for example, an extract of Cistus monsperiensis and ellagitannin contained therein inhibit the Maillard reaction of the skin as a naturally derived material that suppresses saccharification. ..

Further, Patent Document 2 describes an in vivo glycation inhibitor containing soybean saponin B group contained in soybean extract as an active ingredient.

On the other hand, in recent years, an extract of Hamcho has been attracting attention as a material for cosmetics. Hamcho is an annual herb belonging to the Amaranthaceae family and is widely distributed in boreal regions such as Europe, Asia and North America worldwide. It is a halophyte that grows on land in contact with the sea between the average submergence level and the high tide level, which is defined by the tidal tide, and in salt marshes that develop inland.
Hamcho extract has already been incorporated into cosmetics. The purpose of the formulation is skin barrier function recovery action, antioxidant action, anti-inflammatory action (Patent Document 3), stratum corneum cell strengthening action (Patent Document 4), and vitamin C uptake promoting action into the skin (Patent Document 5). The purpose is to exert such effects.
However, it is not known that Hamcho or Hamcho extract has a saccharification reaction inhibitory effect or a Maillard reaction inhibitory effect.

Japanese Patent No. 5441232 Japanese Unexamined Patent Publication No. 2012-17270 Japanese Patent No. 3987825 Japanese Unexamined Patent Publication No. 2010-13373 Japanese Unexamined Patent Publication No. 2018-197211

The present inventor has found a strong saccharification reaction inhibitory effect on Hamcho extract in the process of searching for a substance that suppresses the saccharification reaction of human skin, and made the present invention.
That is, it is an object of the present invention to provide a composition for suppressing a protein saccharification reaction in vivo containing a Hamcho extract as an active ingredient.

The present invention has the following configuration.
(1) A composition for suppressing a protein saccharification reaction in vivo, which contains an extract of Hamcho as an active ingredient.
(2) The composition according to (1), wherein the protein is a skin stratum corneum protein.
(3) The composition according to (1) or (2), wherein the extract of Hamcho is an extract of Hamcho whole plant by the supercritical method.
(4) The composition according to any one of (1) to (3), which contains an extract of Hamcho containing 0.1 to 5% by mass of sterols as an active ingredient.

INDUSTRIAL APPLICABILITY The present invention provides a composition for suppressing a protein glycation reaction in vivo, which comprises an extract of Hamcho as an active ingredient. When applied externally to the skin, the composition of the present invention suppresses the glycation reaction of proteins in the stratum corneum of the skin and suppresses the accumulation of AGEs in the skin tissue.

It is a graph which shows the result of the preliminary test performed in order to determine the optimum concentration of glyoxal used for a saccharification test. AG in the figure is an abbreviation for aminoguanidine. It is a graph which shows the test result which screened the glycation reaction inhibitory effect with respect to a plant extract. It is a graph which shows that the effect of the saccharification reaction inhibitory action differs depending on the extraction method of Hamcho extract.

The present invention will be specifically described.
The "composition" as used in the present invention refers to a composition containing an extract of Hamcho and having an action of suppressing a protein saccharification reaction. The "composition" refers to a cosmetic, a pharmaceutical product, or a quasi-drug containing a Hamcho extract in a dosage form for internal and / or external use together with a known pharmaceutical additive.
Further, the "saccharification reaction" referred to in the present invention is also called a glycation, but is a browning reaction known as an aminocarbonyl reaction or a Maillard reaction. It consists of two stages, an early stage and a late stage, and refers to a reaction in which an amino group of a protein reacts non-enzymatically with an aldehyde group or a ketone group of a reducing sugar to produce an Amadori rearrangement product via a Schiff base. This reaction is reversible. In the late stage, advanced glycation end products (AGEs) are produced through a complex reaction involving the generation of active oxygen. This reaction is irreversible.
"Suppression of saccharification reaction" may be inhibition of the saccharification reaction process at any stage of the above process, and means that the amount of advanced glycation end products (AGEs) produced is suppressed.

The Hamcho used as a raw material of the present invention is not particularly limited in its origin and origin, but is an extract of the whole Salicornia (Salicornia europaea) growing on the coasts of Europe, Asia and North America. Is preferably used.

The extract of Hamcho is used for extraction by washing the Hamcho with water in advance to remove foreign substances, if necessary, and then cutting or pulverizing the part to be extracted as it is or after drying it, if necessary.

In the present invention, a supercritical extract of Salicornia whole plant is preferably used. In particular, supercritical extraction with carbon dioxide is preferable. There are no particular restrictions on the supercritical extraction device. In addition, supercritical extraction may be performed directly from the Salicornia plant. In the extraction operation, the whole Salicornia herb is stored in the extraction tank in the supercritical extraction device, and carbon dioxide is used as the extraction solvent to perform the extraction operation. Suitable extraction conditions are, for example, a carbon dioxide flow rate of 10 to 100 g / min, an extraction pressure of 10 to 100 MPa, and an extraction temperature of 32 to 100 ° C. More preferably, the carbon dioxide flow rate is 65 g / min, the extraction pressure is 25 MPa, and the extraction temperature is 40 ° C. The characteristic of the supercritical extract of Hamcho thus obtained is that it contains a large amount of sterols and terpenes. Here, when a large amount of sterols is contained, the content of sterols in the Hamcho extract is 0.1 to 5.0% by mass, more preferably 1.0 to 1.5% by mass. As long as the extraction method has a sterol content of 0.1 to 5.0% by mass, it is not necessary to use the supercritical extraction method, and the obtained Hamcho extract is preferably used as the Hamcho extract of the present invention. it can.

The Hamcho extract extracted by supercritical extraction is commercially available as a Hamcho extract containing it, and this may be used in the present invention. As such a commercially available product, “Salicornia extract 4% by mass content product: Hydrasarinol” manufactured by CODIF RECHERCHE ET NATURE (sold by Ikunos Co., Ltd.) can be exemplified. Hydrasarinol consists of Hamcho extract 4% by weight and tri (capryl / capric acid) glyceryl 96.0% by weight. Tri (capryl / capric acid) glyceryl is a diluting solvent.

The extract of Hamcho of the present invention suppresses saccharification in the stratum corneum of the skin when applied or applied externally to human or mammalian skin. Therefore, it can be used as a quasi-drug, a drug, etc. for the purpose of preventing or improving the progress of saccharification in the skin.

When the Hamcho extract is used as a preparation for quasi-drugs, pharmaceuticals, or external preparations such as cosmetics, the Hamcho extract is mixed and dispersed by a general manufacturing method directly or with a carrier that is acceptable in the formulation. After that, it can be easily obtained by processing it into a desired form. In this case, in addition to the extract used in the present invention, oily bases such as vegetable oils and animal oils commonly used in such forms, analgesic and anti-inflammatory agents, analgesics, bactericidal disinfectants, astringents, hormones, vitamins , Moisturizers, UV absorbers, alcohols, chelating agents, pH adjusters, preservatives, thickeners, pigments, fragrances and the like can be appropriately blended as long as they do not interfere with the effects of the present invention.
The content of the composition containing the Salicornia extract of the present invention is not particularly limited, but the extract is usually 0.0001 to 10% by mass, preferably 0.001 to 1% by mass of the total composition. The composition is contained in the range of 5% by mass, particularly preferably 0.001 to 1% by mass.

An example of a saccharification reaction suppression test will be shown, and the present invention will be further described.
1. 1. Preliminary test (determination of optimal concentration of glycation reaction inhibitor using skin stratum corneum)
(1) Method An adhesive tape (cellophane tape, made by Nichiban) was brought into close contact with the skin surface of the inner part of the forearm of an adult, and the skin stratum corneum was collected by peeling it off.
The adhesive tape with the stratum corneum was attached to APS-coated slide glass (manufactured by Matsunami Glass Industry Co., Ltd.) and immersed in xylene (manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.) overnight. Then, it was taken out from xylene and washed by immersing it in a phosphate buffered saline (hereinafter referred to as "PBS", manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.) having a pH of 7.4 (room temperature, 5 minutes, 3 times). Then, it was immersed in a glyoxal solution (saccharification inducer; manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.) prepared with ethanol (manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.) to induce saccharification at 37 ° C. overnight. Alternatively, it was immersed in an aminoguanidine (glycation reaction inhibitor; manufactured by Tokyo Chemical Industry Co., Ltd.) solution prepared with ethanol (manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.) and reacted at 37 ° C. overnight.
The glyoxal solution was prepared to have a final concentration of 0,0.625, 1.25, 2.5, 5 mmol / L.
Aminoguanidine (AG) was prepared so that the final concentration was 0, 1, 5, 10 mmol / L.
After inducing saccharification with glyoxal, it was washed with PBS (room temperature, 5 minutes, 3 times), and then hydrogen peroxide (Fujifilm Wako Pure Chemical Industries, Ltd.) was prepared with PBS to a concentration of 3% by volume to block endogenous enzymes. Was immersed in (manufactured by) for 15 minutes at room temperature. Then, it was washed with PBS (room temperature, 5 minutes, 3 times), air-dried, and then photographed with a fluorescence microscope at an excitation wavelength of 360/40 nm and an absorption wavelength of 460/50 nm, 3 fields per tape.
The stratum corneum is known to have autofluorescence, which is known to increase with the accumulation of glycated proteins. Therefore, the fluorescence intensity per stratum corneum cell in the image was measured using image analysis software and used as the amount of glycated protein. The amount of glycated protein was shown as a relative value (ratio / 0 mM GO) when the value of 0 mM glyoxal was 1.

(2) Results The test results are shown in Fig. 1. In FIG. 1, for example, 0 mM AG is a solution of 0 mmol / L, and 1 mM AG is a solution of 1 mmol / L.
As shown in FIG. 1, it was found that autofluorescence increased depending on the concentration of the glycation inducer glyoxal.
This indicates that glyoxal induced protein glycation, accumulated AGEs, and increased autofluorescence.
Glyoxal increased autofluorescence at any of the concentrations evaluated in this study, but 5 mmol / L, which has a high rate of increase in autofluorescence and is considered to be observable even when saccharified, has an inhibitory effect on glycation reaction. It was considered to be the most suitable for evaluation. This concentration was used in the following tests.
In addition, the fluorescence intensity decreased depending on the concentration of aminoguanidine (AG) added.
It was found that aminoguanidine (AG) strongly suppresses glycation by glyoxal at a concentration of 5 mmol / L or more. Therefore, in the following test examples, it was used as a positive control for suppressing the glycation reaction.

2. Screening for glycation reaction suppression effect of plant extract <Primary screening using saccharification reaction suppression test of plant extract>
Primary screening was performed using a commercially available plant extract as a raw material for cosmetics.
The primary screening used was as shown in Table 1 below. The effect of suppressing the protein glycation reaction is already known for the cistus monsperiensis extract.

However, all compositions are listed in% by mass.

The Hamcho extract shown in Table 1 (“Hacho supercritical extract containing 4% by mass: Hydrasarinol” manufactured by CODIF RECHERCHE ET NATURE (sold by Ikunos Co., Ltd.)) was analyzed by a conventional method and found in the Hamcho extract. 1.35% by mass was sterols. The remaining 98.65% by mass of the components are unknown except for the unsaponified fraction. Further, when a plurality of lots were analyzed in the same manner, the total content of sterols was in the range of 0.1 to 5.0% by mass.

(1) Screening test method PBS, positive control aminoguanidine (AG), tri (caprylic acid / capric acid) glyceryl (diluent) commonly contained in each test, and the test in Table 1 are described above. The test was carried out in the same manner as the preliminary test of. Glyoxal was prepared so that the final concentration was 5 mmol / L, and the plant extract, which was the test substance, had a final concentration of 20% by volume, and aminoguanidine (AG) was adjusted to a final concentration of 5 mmol / L as a positive control. Prepared in.

(2) Results Fig. 2 shows the test results.
The test results were shown as the ratio (5 mM / 0 mM ratio) of the fluorescence intensity when saccharification was induced (when glyoxal was 5 mmol / L) and the fluorescence intensity when saccharification was not induced (when glyoxal was 0 mmol / L). For comparison, the values corrected using the following formula are shown. The smaller this value is, the stronger the saccharification inhibitory effect is.

Calculation formula vs Control = (5 mM / 0 mM ratio of subject) / (5 mM / 0 mM ratio of diluent)

The measurement results are shown in FIG. 2 in the following order.
PBS
Aminoguanidine (glycation inhibitor)
Tri (capryl / capric acid) glyceryl (diluent)
Hamcho Extract Sistus Monsperiensis Extract Fernaceae Haz Extract Helichrysum Itaricum Extract

It was confirmed that the cistus monsperiensis extract and the Hamcho extract have a strong inhibitory effect on the glycation reaction of proteins.

3. 3. Confirmation test of saccharification reaction suppression effect by different extraction method of Hamcho extract <Effect test of extract by supercritical extraction method and solvent extraction method>
The Akeshisou extract was tested using extracts with different extraction methods (carbon dioxide supercritical extract, water extract, ethanol extract, 1,3-butylene glycol (BG) extract).

・ Carbon dioxide supercritical extract: “Hydrasarinol” made by CODIF RECHERCHE ET NATURE Akkeshiso supercritical extract 4% by mass content ・ Water extract: Akkeshiso water extract 2% by mass content ・ Ethanol extract: Akkeshiso ethanol extract Product containing 2% by mass ・ BG extract: Product containing 2% by mass of Akkeshiso BG extract

Table 2 shows the compositions of the above four raw materials containing the extracts having different extraction methods.

However, all compositions are listed in% by mass.

(1) Test method The test was conducted by the same test method as the screening test described above. The final concentration of glyoxal was 5 mmol / L, and the Hamcho extract obtained by each extraction method as a test substance was prepared to contain 0.8% by mass as a Hamcho extract at the final concentration. As a positive control, aminoguanidine was prepared to a final concentration of 5 mmol / L and used in the test.
Further, regarding tri (caprylic acid / capric acid) glyceryl contained in hydra linoleic acid in an amount of 96% by mass, and BG contained in each raw material containing a water extract, an ethanol extract, and a BG extract in an amount of 30% by mass. Was measured in the same manner.

(2) Test results Figure 3 shows the test results.
The test result is the ratio of the fluorescence intensity when saccharification is induced (when glyoxal is 5 mmol / L) to the fluorescence intensity when saccharification is not induced (when glyoxal is 0 mmol / L), as in the above cleaning test. 5 mM / 0 mM ratio) was calculated. In the comparison, since the solvent differs depending on the extraction method, the corrected value is shown using the following calculation formula. This value indicates that the smaller it is, the stronger the saccharification inhibitory effect.

Calculation formula vs Control = (5 mM / 0 mM ratio of subject) / (5 mM / 0 mM ratio of control (tri (caprylic acid / capric acid) glyceryl or BG))

No anti-glycation effect was confirmed in each extract of water, ethanol and BG. On the other hand, it was found that the supercritical extract has a saccharification reaction inhibitory effect.

The Hamcho extract contained in Hydrasarinol used in this test is a carbon dioxide supercritical extract and contains 0.1 to 5.0% by mass of sterols. It is generally known that a large amount of minerals, sugars and amino acids are extracted from water extracts, and sterols are hardly extracted. Conceivable.

The following are examples of prescriptions for external creams, external emulsions, and cosmetics containing Hamcho extract. In this formulation example, the blending amount of Hamcho extract may be appropriately increased or decreased depending on the purpose. The Hamcho extract in the formulation is a carbon dioxide supercritical extract.
Formulation Example 1 External cream An external cream having an anti-glycation effect was produced by the following formulation (unit: mass%).
(1) Stearyl alcohol 6
(2) Stearic acid 2
(3) Hydrogenated lanolin 4
(4) Squalan 9
(5) Octyldodecanol 10
(6) POE (25) Cetyl alcohol ether 3
(7) Glycerin monostearate 2
(8) Hamcho extract 5
(9) Preservatives Appropriate amount (10) 1,3 Butylene glycol 6
(11) PEG1500 4
(12) Residual of purified water [Manufacturing method] The above components (1) to (8) are heated and dissolved at 80 ° C. to prepare an oil phase. Ingredients (9) to (12) are heated and dissolved at 70 ° C. to prepare an aqueous phase. The aqueous phase was gradually added to the oil phase to emulsify, and the mixture was cooled to 40 ° C. with stirring and further stirred and cooled to 30 ° C. to obtain an external cream having an anti-glycation effect.

Formulation Example 2 External emulsion An external emulsion having an anti-glycation effect was produced by the following formulation (unit: mass%).
(1) Dipropylene glycol 9
(2) Hamcho extract 0.0001
(3) (Hydroxyethyl acrylic acid / acrylic dimethyl taurine Na) copolymer
0.188
(4) Squalane 0.127
(5) Polysorbate 60 0.028
(6) Di (phytosteryl / octyldodecyl) lauroyl glutamate 1
(7) Glycerin 5
(8) Dimethicone 3
(9) Purified water 76.3919
(10) Carbomer 0.2
(11) Betaine 2
(12) Ethanol 3
(13) Potassium hydroxide 0.065
[Manufacturing method] The above components (2), (4) to (6) and (8) are heated and dissolved at 80 ° C. to prepare an oil phase. The components (1), (3), (7), (9) to (11) are heated and dissolved at 70 ° C. to prepare an aqueous phase. The aqueous phase is gradually added to the oil phase to emulsify, (13) is added to a part of (9) by stirring, and the mixture is cooled to 30 ° C. with stirring. Further, the component (12) was added to obtain an external emulsion having an anti-glycation effect.

Prescription example 3 Ointment
An ointment having an anti-glycation effect was produced according to a conventional method according to the following formulation (unit: mass%).
(1) White petrolatum 24
(2) Propylene glycol 12
(3) Stearyl alcohol 20
(4) Methyl paraoxybenzoate 0.05
(5) Hamcho extract 10
(6) Fragrance 0.1
(7) Purified water 33.85

Formulation Example 4 Cleansing oil A cleansing oil having an anti-glycation effect was produced by the following formulation (unit: mass%) according to a conventional method.
(1) Cetyl ethylcaproate 71.349
(2) Diphenylsiloxyphenyltrimethicone 5
(3) Meadowfoam oil 1
(4) Hexyldecyl isostearate 2
(5) Decaglyceryl diisostearate 8
(6) Hexaglyceryl dicaprate 9
(7) Polyoxyethylene sorbitan monostearate (20EO) 1.7
(8) Diglyceryl monoisostearate 0.4
(9) (Behenic acid / icosanedioic acid) Glyceryl 1.3
(10) Inulin stearate 0.15
(11) Tocopherol 0.1
(12) Hamcho extract 0.001

Formulation Example 5 Cleansing oil A cleansing oil having an anti-glycation effect was produced by the following formulation (unit: mass%) according to a conventional method.
(1) Cetyl ethylcaproate 66.35
(2) Diphenylsiloxyphenyltrimethicone 5
(3) Meadowfoam oil 1
(4) Hexyldecyl isostearate 2
(5) Decaglyceryl dioleate 8
(6) Hexaglyceryl dicaprate 9
(7) Polyoxyethylene sorbitan monostearate (20EO) 1.7
(8) Diglyceryl monoisostearate 0.4
(9) (Behenic acid / icosanedioic acid) Glyceryl 1.3
(10) Inulin stearate 0.15
(11) Tocopherol 0.1
(12) Hamcho extract 5

Prescription example 6 Cleansing gel
According to a conventional method, a cleansing gel having an anti-glycation effect was produced by the following formulation (unit: mass%).
(1) Cetyl ethylcaproate 54.2
(2) Triethylhexanoin 20
(3) Decaglyceryl diisostearate 8
(4) Hexaglyceryl dicaprate 9
(5) Polyoxyethylene sorbitan monostearate (20EO) 1.7
(6) Diglyceryl monoisostearate 0.6
(7) (behenic acid / icosanedioic acid) glyceryl 3
(8) (Palmitate / Ethylcaproic acid) Dextrin 2
(9) Dextrin palmitate 0.5
(10) Hamcho extract 1

Formulation Example 7 Moisturizing beauty emulsion A moisturizing beauty emulsion having an anti-glycation effect was produced by the following formulation (unit: mass%).
(1) Glycerin 10
(2) 1,3-butylene glycol 5
(3) Betaine 3
(4) Dipropylene glycol 7
(5) Citric acid 0.01
(6) Sodium citrate 0.1
(7) Xanthan gum 0.08
(8) 1,2-Pentanediol 1
(9) Squalan 3
(10) Dimethicone 1
(11) Polysorbate 60 0.8
(12) Sorbitan stearate 0.3
(13) Hamcho extract 0.001
(14) Residual of purified water (manufacturing method)
Component 7 heated and dispersed in component 8 is added to component 14, and components 1 to 6 are also added and dissolved by stirring (aqueous phase). An emulsified composition (moisturizing cosmetological emulsion) having an anti-glycation effect was prepared by adding the previously prepared aqueous phase to the heat-dissolved components 9 to 13.

Claims (4)

A composition for suppressing a protein glycation reaction in vivo, which contains an extract of Hamcho as an active ingredient. The composition according to claim 1, wherein the protein is a skin stratum corneum protein. The composition according to claim 1 or 2, wherein the extract of Hamcho is an extract of Hamcho whole plant by the supercritical method. The composition according to any one of claims 1 to 3, which contains an extract of Hamcho containing 0.1 to 5% by mass of sterols as an active ingredient.