Methods, compositions and preparations using young apple extract

Abstract

To provide novel applications of an apple young fruit extract.SOLUTION: Provided is a method for recovering bound water and / or free water of epidermis and / or stratum corneum keratin, using a composition containing an apple young fruit extract. Provided is a skin composition containing an apple young fruit extract. Provided are an inhibitor and a restorer of changes in the α-helix structure of epidermal and / or stratum corneum keratin, containing an apple young fruit extract as an active ingredient. Provided is a water retention-improving agent for epidermis and / or stratum corneum, containing an apple young fruit extract as an active ingredient.SELECTED DRAWING: None

Description

[Technical Field]

[0001] The present invention relates to methods, compositions, and agents using young apple extracts. [Background technology]

[0002] Apples are a highly nutritious food and are enjoyed worldwide. In apple cultivation, fruit thinning is carried out to harvest fruit with high commercial value, resulting in many fruits being picked while still immature. Immature fruits are not suitable for consumption due to their strong sour and astringent taste, and are sometimes discarded. From the perspective of effective resource utilization, attempts are being made to utilize young apples industrially (see Patent Document 1). [Prior art documents] [Patent documents]

[0003] [Patent Document 1] Japanese Patent Application Laid-Open No. 2013-209302 Summary of the Invention [Problem to be solved by the invention]

[0004] It is believed that young apples have great potential for further industrial use. An object of the present invention is to provide a new use of young apple extract. [Means for solving the problem]

[0005] The gist of the present invention is as follows. The methods of the present invention are methods for restoring bound water and / or free water in epidermal and / or stratum corneum keratin, methods for inhibiting changes in the α-helical structure of epidermal and / or stratum corneum keratin, and methods for restoring changes in the α-helical structure of epidermal and / or stratum corneum keratin, all of which use a composition containing young apple fruit extract. The composition of the present invention is a skin composition containing the young apple extract used in the method of the present invention. The agent of the present invention is an agent for inhibiting changes in the α-helical structure of epidermal and / or stratum corneum keratin, an agent for restoring changes in the α-helical structure of epidermal and / or stratum corneum keratin, and an agent for improving the moisture retention ability of the epidermis and / or stratum corneum, which contains young apple fruit extract as an active ingredient. [Effects of the Invention]

[0006] According to the present invention, a new use of young apple extract can be provided. [Brief explanation of the drawings]

[0007] [Figure 1] Figure 1 shows a chart of θ (mdeg) versus wavelength (nm) obtained by measuring the CD spectrum of a control A sample (no young apple extract added) in a CD spectrum measurement experiment. The crosses and the curves connecting them represent the chart before heating, while the black circles and the curves connecting them represent the chart after heating and cooling. [Figure 2] 2 is a chart of θ (mdeg) versus wavelength (nm) when CD spectrum measurement was performed on the sample of Example A (with young apple extract added) in a CD spectrum measurement experiment. The crosses and the curve connecting them show the chart before heating, and the black circles and the curve connecting them show the chart after heating and cooling. [Figure 3] Figure 3 shows the change in θobs (θ222,obs) (mdeg) at a wavelength of 222 nm before heating and after heating and cooling in the charts of the control A sample and the example A sample shown in Figures 1 and 2. [Figure 4] 4 is a chart of θ222,obs (mdeg) versus temperature (°C) when CD spectra were measured for a Control A sample (no young apple extract added) and an Example A sample (young apple extract added) in a CD spectrum measurement experiment. The crosses and the curve connecting them indicate the chart for the Control A sample, and the black circles and the curve connecting them indicate the chart for the Example A sample. [Figure 5] FIG. 5 is a graph showing the peak areas (arbitrary units) obtained by microscopic IR measurement of an untreated sample, a control B sample (no young apple extract added), and an example B sample (young apple extract added) in a microscopic IR measurement experiment. DETAILED DESCRIPTION OF THE INVENTION

[0008] Hereinafter, a form for implementing the present invention (hereinafter referred to as "the present embodiment") will be described in detail with reference to the drawings. However, the present invention is not limited to the following description and can be implemented in various modified forms within the scope of its gist.

[0009] (method) The method of this embodiment is a method using a composition containing young apple extract, and more specifically, may be a method for restoring bound water and / or free water in epidermal and / or stratum corneum keratin, a method for inhibiting changes in the α-helical structure of epidermal and / or stratum corneum keratin, or a method for restoring changes in the α-helical structure of epidermal and / or stratum corneum keratin. The method of this embodiment is used outside of medical practice. The composition used in the method of this embodiment may be the composition of this embodiment described below.

[0010] The young apple extract used in the method of this embodiment may be an extract from young apples of various varieties (scientific name: Malus pumila), which are plants of the genus Malus in the family Rosaceae.

[0011] Here, the term "variety" of apples refers to any variety, and may be an early-season, mid-season, or late-season variety. Examples of varieties include Fuji, Tsugaru, Ourin, Jonagold, Hokuto, Mutsu, Chiaki, Delicious, Kogyoku, Sansa, Akiakari, Youkou, Shinano Sweet, Sekaiichi, Asahi, Indo, Granny Smith, Kunimitsu, Star King, Tsugaru, and Hack Nine.

[0012] The term "young fruit" refers to an immature fruit at any stage after fruit set but before ripening. For example, apple fruit thinning is usually carried out in May or June, approximately 20 to 45 days after fruit set. Young fruits with a diameter of approximately 2 to 5 cm that are picked can be included in the "young fruit" of this embodiment. In this case, the "young fruit" may include all or part of the immature fruit, specifically the peel, pulp, puree, juice, and seeds.

[0013] The "extract" may be an extract obtained by soaking young apples in an extraction solvent and removing solids from the resulting extract, or an extract obtained by removing solids from the juice contained in young apples. The "extract" may be an extract obtained by concentrating or drying the extract or juice from which the solids have been removed, or may be a solid obtained by drying a liquid, or a powder obtained by drying using a method such as spray drying.

[0014] The following describes an example of a method for extracting from young apples. The young apples used for extraction may be fresh or processed. They may be used immediately after thinning, or may be used some time after thinning as long as they are not decayed. Furthermore, the young apples may be pretreated, such as by cutting or crushing, before being used for extraction to ensure efficient extraction. There are no particular limitations on the pretreatment, but cryo-crushing or freeze-crushing is preferred. The extraction solvent is not particularly limited, but examples include water, lower alcohols (methanol, ethanol, 1-propanol, 2-propanol, 1-butanol, 2-butanol, etc.), polyhydric alcohols (glycerin, propylene glycol, 1,3-butylene glycol (BG), etc.), ketones (acetone, methyl ketone, etc.), ethers (diethyl ether, tetrahydrofuran, etc.), esters such as ethyl acetate, and hexane. Among these, water, lower alcohols, and polyhydric alcohols are preferred in terms of usability and safety. These may be used alone or in combination of two or more. The extraction conditions can be appropriately designed by a person skilled in the art; for example, young apples may be immersed in an extraction solvent at room temperature (1 to 30°C) or at elevated temperatures (50 to 80°C) for a period of 1 hour to several months, with low-temperature conditions (1 to 15°C) for 1 to 10 days being particularly preferred. The mass ratio of young apples to extraction solvent may be appropriately designed, and for example, the lower limit of the amount of extraction solvent is not particularly limited as long as it is an amount that allows the raw material to be sufficiently immersed per 100 parts by mass of young apples, and is preferably 50 parts by mass or more, more preferably 100 parts by mass or more, and even more preferably 200 parts by mass or more. The upper limit of the amount of extraction solvent can also be determined from an economical perspective, and can be, for example, 10,000 parts by mass or less, preferably 5,000 parts by mass or less, and more preferably 1,000 parts by mass or less per 100 parts by mass of raw material. When water is used as the extraction solvent, the extraction may be carried out by steam distillation. Solid matter may be removed by a conventional method. The solids concentration of the resulting young apple extract after removing the solvent may be 0.1 to 10% by mass.

[0015] The following describes an example of a method for using the juice of young apples. The juice may be obtained by crushing washed young apples as the raw material at a pH of 3.2 to 4.6, preferably 3.4 to 4.3 (pH measurement temperature: 20°C), treating the resulting crushed material with pectinase for clarification, and removing solids.

[0016] Compositions containing young apple extract effectively maintain the bound water and / or free water contained in the epidermis and / or stratum corneum. The inventors have found that young apple extract has the effect of restoring the bound water and / or free water in epidermal and / or stratum corneum keratin. Young apple extract can also inhibit the loss of bound water and / or free water in epidermal and / or stratum corneum keratin, as well as restore the total amount of water in epidermal and / or stratum corneum keratin and inhibit the loss of the total amount of water. The mechanism by which the above-mentioned effects on bound water and / or free water are exerted is thought to be as follows. In animals, including humans and non-humans, the epidermis and stratum corneum of skin contain keratin, a protein that constitutes the cytoskeleton. The two-dimensional structure of keratin is primarily α-helical, and maintaining the amount of α-helical structure is crucial for maintaining the proper cytoskeleton of the epidermis and / or stratum corneum. The inventors have obtained results suggesting that young apple extract has the effect of stabilizing the hydration structure of keratin, thereby facilitating the maintenance of the α-helical structure. More specifically, as described in the Examples below, young apple extract has the effect of inhibiting changes in the α-helical structure of human epidermal-derived keratin and also of recovering from such changes. The above findings of the inventors can be said to be new discoveries made through the present invention.

[0017] (composition) The composition of the present embodiment contains an extract of young apple fruits. The young apple extract used in the composition of this embodiment may be the same as the young apple extract used in the method of this embodiment.

[0018] The composition of this embodiment may contain additives in addition to the young apple extract, as needed.

[0019] Such additives include cell activators, antioxidants, moisturizers, UV protection agents, solvents (water, alcohols, etc.), oils, surfactants, thickeners, powders, chelating agents, pH adjusters, emulsifiers, stabilizers, colorants, glossing agents, flavoring agents, odor masking agents, excipients, binders, disintegrants, lubricants, diluents, osmotic pressure adjusters, fragrances, etc.

[0020] The composition of the present embodiment may be prepared in various forms, such as a liquid, emulsion, cream, solid, gel, or paste. The composition may also be prepared in various dosage forms, such as an oil-based system, a water-in-oil emulsion system, or an oil-in-water emulsion system. Specifically, the composition can be various cosmetics, such as skin care cosmetics such as lotion, emulsion, cream, serum, cosmetic oil, lip balm, hand cream, facial cleanser, and cleanser; makeup cosmetics such as foundation, makeup base, blush, eye shadow, mascara, eyeliner, eyebrow pencil, overcoat, lipstick, and lip gloss; scalp or hair cosmetics such as hair tonic, hair cream, shampoo, rinse, conditioner, and hair styling products; and massage cosmetics.

[0021] The composition of the present embodiment may be used as a cosmetic, a quasi-drug, or a pharmaceutical.

[0022] The method for producing the composition of the present embodiment is not particularly limited, and may be a method commonly used in the art. The composition of this embodiment is preferably used in the method of this embodiment.

[0023] (agent) The agent of this embodiment contains young apple fruit extract as an active ingredient, and examples of the agent include an agent for inhibiting changes in the α-helix structure of epidermal and / or stratum corneum keratin, an agent for restoring changes in the α-helix structure of epidermal and / or stratum corneum keratin, an agent for improving the moisture retention ability of the epidermis and / or stratum corneum, an agent for restoring damage to the epidermis and / or stratum corneum, and an agent for inhibiting damage to the epidermis and / or stratum corneum. As mentioned above, and as will be described in the Examples below, young apple extract has the effect of restoring and suppressing changes in the α-helical structure of keratin derived from human epidermis, and is therefore expected to have the effect of recovering from and / or suppressing stimuli and damage such as heat, oxidation, and ultraviolet rays by suppressing and recovering changes in the α-helical structure of epidermal and / or stratum corneum keratin.

[0024] The agent of this embodiment may be the composition of this embodiment.

[0025] In this embodiment, the young apple extract can also be used to produce the above-mentioned agents of this embodiment. [Example]

[0026] The present invention will be described in more detail below with reference to examples, but the present invention is not limited to the following examples in any way.

[0027] The materials used in the examples and comparative examples described below are listed below. [Young apple extract] Young (immature) fruits of the Hakunai variety grown in Hokkaido, measuring up to 20 mm in diameter and grown within one month of flower abscission, were added with a 13-fold volume of 20% (w / w) 1,3-butylene glycol aqueous solution and extracted at low temperatures (1-15°C) for two days. After removing the solvent, the solid concentration was 0.3% by mass.

[0028] <1. CD spectrum measurement> <Measurement method> Measurement equipment: a circular dichroism spectrometer J-720 (JASCO Corporation) and a Peltier-type thermostatic cuvette holder PTC-423L (JASCO Corporation) were used, along with a quartz cell for spectrophotometers with an optical path length of 1 mm. The measurement conditions were as follows: Measurement cell: quartz 1mm cell Measurement wavelength: 190~250nm Sensitivity: Standard Temperature range: 20.0 to 98.0°C Heating rate: 1.0℃ / min Measurement sample: Human epidermal keratin (manufactured by SIGMA-ALDRICH) was thoroughly dialyzed against a buffer solution of 5 mM Na2PO4 / NaHPO4, 50 mM NaCl, pH 7.4, and then diluted with the same buffer solution to a measurable concentration. The control A sample was simply diluted with the buffer solution, while the example A sample containing young apple extract had the components added to the solution at 1% by volume. Measurement and analysis conditions: Measurements were performed for each sample according to the procedure below. The regeneration rate of the α-helix structure after heating and cooling was calculated by comparing (1) and (4), and the thermal denaturation temperature was obtained by (2). (1) CD spectrum measurement is performed at 20.0°C at wavelengths of 190 to 250 nm. (2) Fix the wavelength at 222 nm and measure the temperature of the same sample from 20.0 to 98.0°C at a rate of 1.0°C / min. (3) After the measurement at 98.0°C was completed, the sample was left to stand for 10 minutes, and then cooled to 20.0°C using a Peltier temperature controller. (4) After leaving the sample to stand at 20.0°C for 10 minutes, the CD spectrum is measured again at 20.0°C at a wavelength of 190 to 250 nm.

[0029] <Measurement results> Figure 1 shows a chart of θ (mdeg) versus wavelength (nm) obtained by measuring the CD spectrum of a control A sample (no young apple extract added) in a CD spectrum measurement experiment. The crosses and the curves connecting them represent the chart before heating, while the black circles and the curves connecting them represent the chart after heating and cooling.

[0030] 2 is a chart of θ (mdeg) versus wavelength (nm) when CD spectrum measurement was performed on the sample of Example A (with young apple extract added) in a CD spectrum measurement experiment. The crosses and the curve connecting them show the chart before heating, and the black circles and the curve connecting them show the chart after heating and cooling.

[0031] Figure 3 shows the change in θobs (θ222,obs) (mdeg) at a wavelength of 222 nm before heating and after heating and cooling in the charts of the control A sample and the example A sample shown in Figures 1 and 2.

[0032] Native keratin proteins contain a large amount of α-helical structures, and the content of α-helical structures can be determined by the peak intensity at 222 nm in CD measurements.

[0033] As shown in Figure 2, when the peak intensity at 222 nm was compared at the completion of heating (97.6-98.0°C) under measurement and analysis conditions (2) and (4), 84.7% of the peak intensity that decreased during heating was recovered by cooling after heating in Control A, whereas in Example A, the recovery rate increased to 94.5% by adding the young apple extract prepared above (containing 1% by volume of the original solution). In other words, it can be said that approximately 64% of the α-helix structure that would not be regenerated by simple heating and cooling alone was regenerated by adding the young apple extract.

[0034] As shown in FIG. 3, the change in θ222,obs in Example A sample was approximately 34% of the change in θ222,obs in Control A sample. These results indicate that heating in the presence of young apple extract has the effect of inhibiting structural changes in human epidermal keratin, and / or that cooling in the presence of young apple extract has the effect of restoring the structure of human epidermal keratin.

[0035] 4 is a chart of θ222,obs (mdeg) versus temperature (°C) when CD spectra were measured for a Control A sample (no young apple extract added) and an Example A sample (young apple extract added) in a CD spectrum measurement experiment. The crosses and the curve connecting them indicate the chart for the Control A sample, and the black circles and the curve connecting them indicate the chart for the Example A sample. 4 shows thermal denaturation curves obtained by measuring the sample while heating it from 20.0 to 98.0°C at a fixed measurement wavelength of 222 nm. The thermal denaturation temperature (°C) was calculated from the inflection points of the thermal denaturation curves for Control A and the sample of Example A to which young apple extract was added. The result was 39.28°C for Control A and 48.92°C for the sample of Example A to which young apple extract was added. This indicates that the addition of young apple extract improves the thermodynamic stability of the α-helix structure of keratin protein.

[0036] <2. Microscopic IR Measurement> <Measurement method> Measurement equipment: A humidity control device was used at SPring-8 BL43IR to create a humidity environment of 30% RH. Measurement sample: The stratum corneum was peeled from commercially available human skin, and 10mm x 10mm square human skin-derived stratum corneum sheets were treated under the conditions shown below. (1) Untreated sample: No special treatment was performed. (2) (Control B sample): The sample was immersed in a 3% H2O2 aqueous solution (pH 10.4) for 20 minutes to damage it, then washed with water seven times, and then dried in an environment of 25°C for 24 hours. (3) (Example B sample): After the above-mentioned damage treatment, the sample was immersed in an aqueous solution of young apple extract (containing 1% by mass of the original solution) for 3 hours, and then dried in an environment of 25°C for 24 hours. Measurement and analysis conditions: Aperture size was approximately 10 μm × 10 μm, and measurements were taken at 20 points (X = 5, Y = 4) with 10 μm steps (both the vertical and horizontal spacing of each aperture point was 10 μm), 512 accumulations, and baseline correction was performed on the measurement results. The peak area of ​​the amide I mode (wavenumber 1806-1600 cm) derived from the amide bond of the protein was calculated. -1 ) normalized by the total amount of water (wavenumber 3620-3100 cm -1 ), bound water (wave number 3420~3245cm -1 ), free water (wave number 3620~3420cm -1 ) peak area was calculated.

[0037] <Measurement results> FIG. 5 is a graph showing the peak areas (arbitrary units) obtained by microscopic IR measurement of an untreated sample, a control B sample (no young apple extract added), and an example B sample (young apple extract added) in a microscopic IR measurement experiment.

[0038] Table 1 shows the numerical data for each peak area shown in FIG.

[0039] [Table 1]

[0040] Water peaks (peaks derived from OH groups) in IR measurements can be broadly classified into two types: bound water, which is weakly bound to the structure and does not easily increase or decrease due to external influences such as humidity, and free water, which easily increases or decreases due to influences such as humidity. In the Control B sample, which was subjected to oxidative damage treatment, both bound water and free water decreased compared to the untreated sample, but in the Example B sample to which young apple extract was subsequently added, both bound water and free water were shown to recover to levels close to those of the untreated sample. Furthermore, the Example B sample also showed a recovery in total water content compared to the Control B sample. This indicates that the young apple extract essentially improves the moisture content in the stratum corneum, which is reduced by oxidative damage. [Industrial Applicability]

[0041] According to the present invention, a new use of young apple extract can be provided.

Claims

1. A method for restoring bound water lost in oxidatively damaged epidermis and / or stratum corneum keratin to the state before oxidative damage, using a composition containing young apple extract (excluding medical procedures and cases for moisturizing and maintaining moisture).

2. A moisture retention function improver containing young apple extract as an active ingredient, which restores the bound water lost in oxidatively damaged epidermis and / or stratum corneum keratin to the state before oxidative damage (excluding cases of moisturizing or retaining moisture).

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